Crystalline form and synthesis method of 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate

Abstract

The present invention relates to a compound of formula (X): TIFF2023532330000063.tif1738. The present invention also provides pharmaceutical compositions containing polymorphs of the compound, and methods of treating a condition in a subject by administering a polymorph of the compound. The composition may include a crystal of the compound of Formula (X). The crystal can have any of the properties described above for the crystal of the compound. In another aspect, the present invention provides methods of treating a condition in a subject by administering to a subject having or at risk of developing a condition a therapeutically effective amount of a composition containing a polymorph of the compound of Formula (X).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to 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 by reference.

[0002] FIELD OF THE INVENTION The present invention relates to crystallographic forms of 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate and methods for chemically synthesizing said compound. [Background technology]

[0003] background Heart disease is a leading cause of death worldwide, accounting for 15 million deaths worldwide in 2015. In many forms of heart disease, decreased cardiac efficiency results from alterations in mitochondrial energy metabolism. Mitochondria are intracellular compartments where glucose- and fatty acid-derived metabolites are oxidized to produce high-energy molecules. As fatty acid oxidation increases in the heart, glucose oxidation decreases, and vice versa. Although glucose oxidation is a more efficient energy source, fatty acid oxidation predominates in cardiac mitochondria in certain types of heart disease, such as heart failure, ischemic heart disease, and diabetic cardiomyopathy. As a result, the heart's pumping capacity decreases.

[0004] IUPAC name 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate and the following structure: [ka] CV-8972, having the formula: ##STR00002## was recently identified as a promising therapeutic candidate for treating or preventing cardiac conditions due to its pharmacokinetic profile. Summary of the Invention [Means for solving the problem]

[0005] summary Crystallographic forms of CV-8972 and compositions containing them are provided herein. The present invention recognizes that CV-8972 crystals exist in multiple polymorphic forms, and that one polymorph, Form A, is the most stable under ambient temperature and relative humidity conditions. Therefore, CV-8972 Form A crystals are useful for preparing pharmaceutical compositions. For example, pharmaceutical compositions containing Form A polymorph do not require special handling during storage or distribution. Furthermore, such compositions may retain greater efficacy than compositions containing other polymorphs or mixtures of polymorphs. The present invention also provides methods for treating cardiac conditions in subjects using CV-8972 polymorphs, such as Form A.

[0006] The present invention also provides a method for the synthesis of CV-8972. Previous schemes for synthesizing CV-8972 require the formation of the free base form of 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethanol, also known as CV-8814, and the conversion of the free base form of CV-8814 to the hydrochloride salt. In such schemes, CV-8814 must then be converted back to its free base form for coupling to nicotinic acid to form the free base form of CV-8972. The present invention provides a synthetic scheme for CV-8972 that bypasses the reversible conversion of CV-8814 between the free base form and the HCl salt form. In the scheme 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 to nicotinic acid to form CV-8972. The synthetic scheme of the present invention requires fewer steps and is therefore simpler, faster, and provides superior yields than previous methods for making CV-8972.

[0007] In an aspect, the present invention provides a compound of formula (X): [ka] The present invention provides crystals containing polymorphs of the compound of formula (I).

[0008] The polymorph may be Form A, Form B, Form C, Form D or Form E.

[0009] The crystal may be substantially free of one or more other polymorphs. For example, the crystal may comprise polymorph Form A, and may be substantially free of polymorph Form B, Form C, Form D, and Form E.

[0010] The crystal may comprise a hydrochloride salt of the compound of formula (X). The crystal may comprise the compound of formula (X) and hydrochloride ions in a defined stoichiometric ratio. The crystal may comprise the compound and hydrochloride ions in a 1:3 stoichiometric ratio.

[0011] The crystals may include a hydrated form of the compound of formula (X). The crystals may include a monohydrate form of the compound. The crystals may include an anhydrous form of the compound.

[0012] In another aspect, the present invention provides pharmaceutical compositions comprising polymorphs of the compound of formula (X).

[0013] The polymorph may be Form A, Form B, Form C, Form D or Form E.

[0014] The composition may be substantially free of one or more other polymorphs. For example, the composition may comprise the Form A polymorph, and may be substantially free of the Form B, Form C, Form D, and Form E polymorphs.

[0015] The composition may comprise a hydrochloride salt of the compound of formula (X). The composition may comprise the compound of formula (X) and hydrochloride ion in a defined stoichiometric ratio. The composition may comprise the compound and hydrochloride ion in a 1:3 stoichiometric ratio.

[0016] The composition may include a hydrated form of the compound of formula (X). The composition may include a monohydrate form of the compound. The composition may include an anhydrous form of the compound.

[0017] The composition may be formulated for any route or mode of administration. The composition may be formulated for oral, cutaneous, intestinal, intraarterial, intramuscular, intraocular, intravenous, intranasal, oral, parenteral, pulmonary, rectal, subcutaneous, topical, or transdermal administration. The composition may be formulated for administration by injection or using or on an implantable medical device (e.g., a stent or drug-eluting stent or balloon equivalent).

[0018] The composition may be formulated as a single unit dose. The composition may be formulated as divided doses.

[0019] The composition may contain a prescribed dose of the compound. Doses include about 10 mg to about 2000 mg of the compound, 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 50 mg The amount of the active ingredient may be about 100 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. A dose can 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.

[0020] The composition may comprise a crystal of the compound of formula (X). The crystal may have any of the properties described above for a crystal of the compound.

[0021] In another aspect, the present invention provides methods of treating a condition in a subject by administering to a subject having or at risk of developing the condition a therapeutically effective amount of a composition containing a polymorph of the compound of Formula (X).

[0022] The polymorph may be Form A, Form B, Form C, Form D or Form E.

[0023] The composition can have any of the properties described above for compositions comprising the compound, including crystals of the compound of formula (X).

[0024] The compositions may be administered by any suitable route or mode of administration: orally, cutaneously, intestinally, intraarterially, intramuscularly, intraocularly, intravenously, intranasally, orally, parenterally, intrapulmonary, rectally, subcutaneously, topically, transdermally, by injection, or with or on an implantable medical device (e.g., a stent or drug-eluting stent or balloon equivalent).

[0025] The composition may be administered as a single unit dose. The composition may be administered as divided doses.

[0026] The composition may be administered in one dose per day. The composition may be administered in multiple doses per day. The composition may be administered in 2, 3, 4, 5, 6, 8 or more doses per day.

[0027] The composition can contain a prescribed dose of the compound, such as any of the doses described above.

[0028] The dose(s) may be administered over a defined period of time. One or more doses may be administered daily for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks, or longer.

[0029] The condition can be a cardiovascular condition. The cardiovascular condition can be any condition that can be improved by improving cardiac mitochondrial function. The disease, disorder, or condition can be a cardiovascular condition. The disease, disorder, or condition can be an 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 arterial disease, rheumatic heart disease, stroke, transient ischemic attack, or valvular heart disease. Angina pectoris can be refractory to other medical interventions.

[0030] The condition can be a rheumatic condition. The rheumatic condition can be acute kidney injury, alcoholic cardiomyopathy, angina (e.g., refractory angina and angina associated with heart failure), ankylosing spondylitis, autoimmune-related lung disease, Behcet's disease, bursitis, cachexia, myocardial fibrosis, chemotherapy-induced chronic fatigue syndrome, claudication (e.g., peripheral claudication), contrast nephropathy, cyanotic heart disease, dermatomyositis, dilated cardiomyopathy, imbalance, fibromyalgia, weakness, gout, Gulf War syndrome, heart failure, hypertrophic cardiomyopathy, induced nephropathy, It may be infectious arthritis, inflammatory arthritis, inflammatory eye disease, inflammatory myositis, ischemic cardiomyopathy, juvenile idiopathic arthritis, left ventricular dysfunction, lupus, myomyopathies, myofascial pain syndrome, myositis, osteoarthritis, osteonecrosis of the jaw, osteoporosis, polymyalgia rheumatica, polymyositis, psoriatic arthritis, pulmonary arterial hypertension, pulmonary fibrosis, rare muscle diseases, rheumatoid arthritis, sarcoidosis, sarcopenia, scleroderma, Sjögren's syndrome, tendonitis, tinnitus, vasculitis or vertigo.

[0031] The condition can be fibrosis. The fibrosis can be associated with another disease, disorder or condition. For example, fibrosis can include or be associated with adhesive capsulitis, aneurysm, angina pectoris, arteriosclerosis, arthrofibrosis, 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, endomyocardial fibrosis, glial scar, heart attack, heart failure, high blood pressure (hypertension), idiopathic pulmonary fibrosis, ischemic heart disease, keloid, mediastinal fibrosis, myelofibrosis, nephrogenic systemic fibrosis, old myocardial infarction, pericardial disease, peripheral arterial disease, Peyronie'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.

[0032] The condition can be cancer, which can be bladder cancer, brain cancer, breast cancer, carcinoma, cervical cancer, colon cancer, colorectal cancer, gastric cancer, glioblastoma, glioma, head and neck cancer, kidney cancer, leukemia, liposarcoma, liver cancer, lung cancer, lymphoma, medulloblastoma, melanoma, muscle cancer, neuroblastoma, oligoastrocytoma, oligodendroglioma, osteosarcoma, ovarian cancer, pancreatic cancer, paraganglioma, prostate cancer, sarcoma, or thyroid cancer.

[0033] In another aspect, the present invention provides a method of altering cardiac remodeling by administering to a subject having or at risk of developing cardiac remodeling a therapeutically effective amount of a composition containing a polymorph of the compound of formula (X).

[0034] The polymorph may be Form A, Form B, Form C, Form D or Form E.

[0035] The composition can have any of the properties described above for compositions comprising the compound, including crystals of the compound of formula (X).

[0036] The compositions may be administered by any suitable route or mode of administration: orally, cutaneously, enterally, ocularly, intravenously, intranasally, orally, parenterally, pulmonary, subcutaneously, topically, transdermally, by injection, or with or on an implantable medical device (e.g., a stent or drug-eluting stent or balloon equivalent).

[0037] The composition may be administered as a single unit dose. The composition may be administered as divided doses.

[0038] The composition may be administered in one dose per day. The composition may be administered in multiple doses per day. The composition may be administered in 2, 3, 4, 5, 6, 8 or more doses per day.

[0039] The composition can contain a prescribed dose of the compound, such as any of the doses described above.

[0040] The dose(s) may be administered over a defined period of time. One or more doses may be administered daily for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks, or longer.

[0041] Cardiac remodeling may be associated with a disease, disorder, or condition. Cardiac remodeling may be associated with cardiovascular disease. For example, cardiac remodeling may be associated with abnormal subclavian artery, aortic regurgitation, aortic stenosis, arteriovenous malformations and fistulas, atrial septal defect, atrioventricular septal defect, bicuspid aortic valve, cardiomegaly, cardiomyopathy, coarctation of the aorta, complete heart block, concentric hypertrophy, congenital heart defects, congenital heart disease, coronary artery disease, dextrocardia, dextro-transposition of the great arteries, diabetes, diet, double aortic arch, bi-atrioventricular left ventricular insertion, double outlet right ventricle, Ebstein's anomaly, giant hepatic hemangioma, heart failure, high cholesterol, high output hemodialysis fistula, hypertension, high blood pressure, hypoplastic left heart syndrome, hypoplastic right heart syndrome, interrupted aortic arch, aortic aorta. It may be associated with left lateral transposition of the heart, mitral regurgitation (which also causes left atrial volume overload), mitral stenosis, myocardial ischemia, obesity, outflow obstruction, partial anomalous pulmonary venous connection, patent ductus arteriosus, pentalogy of Cantrell, persistent truncus arteriosus, pressure overload, pulmonary atresia, pulmonary hypertension, pulmonary regurgitation, pulmonary stenosis, rhabdomyoma, right ventricular volume overload, Scimitar syndrome, Shawn syndrome, tetralogy of Fallot, total anomalous pulmonary venous connection, transposition of the great arteries, tricuspid atresia, tricuspid regurgitation, tobacco, alcohol, or other drug use, valvular heart disease, ventricular dilation, ventricular hypertrophy, ventricular septal defect, volume overload, and Wolff-Parkinson-White syndrome.

[0042] In another aspect, the present invention provides the use of a crystal containing a polymorph of the compound of formula (X) for preparing a medicament.

[0043] In an embodiment of the use, the polymorph is Form A, Form B, Form C, Form D or Form E.

[0044] In an embodiment of this use, the crystals are substantially free of one or more other polymorphs. In an embodiment of this use, the crystals comprise the Form A polymorph and are substantially free of the Forms B, C, D, and E polymorphs.

[0045] In an embodiment of this use, the crystal comprises the hydrochloride salt of the compound of formula (X). In an embodiment of this use, the crystal comprises the compound of formula (X) and chloride ions in a defined stoichiometric ratio. In an embodiment of this use, the crystal comprises the compound and chloride ions in a 1:3 stoichiometric ratio.

[0046] In an embodiment of this use, the medicament comprises a hydrated form of the compound of formula (X). In an embodiment of this use, the medicament comprises a monohydrate form of the compound. In an embodiment of this use, the medicament comprises an anhydrous form of the compound.

[0047] In an embodiment of this use, the medicament is formulated for oral, cutaneous, intestinal, intraarterial, intramuscular, intraocular, intravenous, intranasal, oral, parenteral, pulmonary, rectal, subcutaneous, topical, or transdermal administration. In an embodiment of this use, the medicament is formulated for administration by injection or with or on an implantable medical device (e.g., a stent or drug-eluting stent or balloon equivalent).

[0048] In an embodiment of this use, the medicament is formulated as a single unit dose. In an embodiment of this use, the medicament is formulated as a split dose.

[0049] In an embodiment of this use, the medicament contains the compound in an amount of 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, or about 50 mg to about 3 00mg, about 50mg to about 200mg, about 100mg to about 2000mg, about 100mg to about 1000mg, about 100mg to about 800mg, about 100mg to about 600mg, about 100mg to about 400mg, about 100mg to about 300mg, about 100mg to about 200mg, about 200mg to about 2000mg, about 200mg to about 1000mg, about 200mg to about 800mg, about 200mg to about 600mg, about 200mg to about 400mg, about 200mg to about 300mg, about 300mg to about 2000mg, about 300mg to about 1000mg, about 300mg to about 800mg, about 300mg to about 600mg, or about 300mg to about 400mg. In an embodiment of this use, the medicament comprises 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.

[0050] In another aspect, the present invention provides a compound of formula (X): [ka] 1. A method for preparing 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol are reacted to give a compound of formula (IX): [ka] generating the free base form of reacting the free base form of the compound of formula (IX) with nicotinic acid to produce a compound of formula (X). This is done by carrying out The present method provides a method that does not involve producing a salt form of the compound of formula (IX).

[0051] The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol can include one or more of a solvent, a catalyst, or other chemicals. The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol can include one or more of sodium triacetoxyborohydride, acetic acid, and 2-methyltetrahydrofuran.

[0052] The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol can be carried out at a specified temperature, such as about 10°C to about 30°C, about 15°C to about 30°C, about 20°C to about 30°C, about 25°C to about 30°C, about 10°C to about 25°C, about 15°C to about 25°C, about 20°C to about 25°C, about 10°C to about 20°C, or about 15°C to about 20°C.

[0053] The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol may be free of a particular solvent, catalyst, or other chemical. The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol may be free of dichloromethane.

[0054] Reacting the free base form of the compound of formula (IX) with nicotinic acid can produce the free base form of the compound of formula (X).

[0055] The step of reacting the free base form of the compound of formula (IX) with nicotinic acid can include one or more of a solvent, a catalyst, or other chemicals. The step of reacting the free base form of the compound of formula (IX) with nicotinic acid can include one or more of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, 4-(dimethylamino)pyridine, and dichloromethane.

[0056] The step of reacting the free base form of the compound of formula (IX) with nicotinic acid can be carried out at a temperature of 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.

[0057] The method may include converting the free base form of the compound of formula (X) into a salt form of the compound of formula (X). The salt form of the compound of formula (X) may be an HCl salt. The salt form of the compound of formula (X) may be a monohydrate.

[0058] The step of converting the free base form of the compound of formula (X) to a salt form of the compound of formula (X) can include one or more of a solvent, a catalyst, or other chemical. The step of converting the free base form of the compound of formula (X) to a salt form of the compound of formula (X) can include one or more of HCl and methyl ethyl ketone.

[0059] The step of converting the free base form of the compound of Formula (X) to the salt form of the compound of Formula (X) can be carried out at a specified temperature. The step of converting the free base form of the compound of Formula (X) to the salt form of the compound of Formula (X) can be carried out at about 40°C to about 60°C, about 45°C to about 60°C, about 50°C to about 60°C, about 55°C to about 60°C, about 40°C to about 55°C, about 45°C to about 55°C, about 50°C to about 55°C, about 40°C to about 50°C, about 45°C to about 50°C, about 40°C to about 50°C, about 40°C, about 45°C, about 50°C, about 55°C, or about 60°C.

[0060] The method can include converting a salt form of the compound of Formula (X) from a first crystalline form to a second crystalline form, wherein the first and second crystalline forms can each independently be Form A, Form B, Form C, Form D, or Form E.

[0061] The step of converting the salt form of the compound of Formula (X) from a first crystalline form to a second crystalline form may include one or more of the steps of changing the solvent of the salt form of the compound of Formula (X) and incubating the salt form of the compound of Formula (X) at about 60°C.

[0062] The process may be carried out without the use of one or more of a solvent, catalyst or other chemicals. The process may be carried out without the use of one or more of dioxane, ethyl acetate or potassium carbonate.

[0063] The method may include purifying the free base form of the compound of formula (IX).The method may include crystallizing the free base form of the compound of formula (IX).

[0064] In another aspect, the present invention provides a method for producing a pharmaceutical composition comprising: Compounds of formula (1): [ka] with a compound of formula (2): [ka] to produce the free base form of the compound of formula (IX) [ka] generating a The free base form of the compound of formula (IX) can be converted into a compound of formula (3): [ka] to produce the free base form of the compound of formula (X), and Converting the free base form of the compound of formula (X) to the HCl salt of the compound of formula (X). A method for preparing a compound of formula (X) by carrying out The present method provides a method that does not involve producing a salt form of the compound of formula (IX).

[0065] The method may include purifying the free base form of the compound of formula (IX).The method may include crystallizing the free base form of the compound of formula (IX). In an embodiment of the present invention, for example, the following items are provided: (Item 1) Compounds of formula (X): [ka] Crystals containing the Form A polymorph of (Item 2) Item 1, wherein the crystal comprises a hydrochloride salt of the compound. (Item 3) 3. The crystal of claim 2, wherein the crystal comprises a hydrated form of the compound. (Item 4) 4. The crystal according to item 3, wherein the hydrated form of the compound is a monohydrate. (Item 5) 2. The crystal according to item 1, wherein the crystal is substantially free of Form B, Form C, Form D and Form E polymorphs. (Item 6) Compounds of formula (X): [ka] A pharmaceutical composition comprising the Form A polymorph of (Item 7) 7. The composition of claim 6, wherein the composition comprises the hydrochloride salt of the compound. (Item 8) 8. The composition of claim 7, wherein the composition comprises a hydrated form of the compound. (Item 9) 9. The composition of claim 8, wherein the hydrate form of the compound is a monohydrate. (Item 10) 7. The composition of claim 6, wherein the composition is substantially free of Form B, Form C, Form D and Form E polymorphs. (Item 11) 7. The composition of claim 6, wherein the composition is formulated for oral administration. (Item 12) 7. The composition of claim 6, wherein the composition is formulated as a single unit dosage. (Item 13) 7. The composition of claim 6, wherein the composition is formulated as a divided dose. (Item 14) A subject having or at risk of developing a condition is administered a therapeutically effective amount of a compound of formula (X):

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[0066] [Figure 1] FIG. 1 is a space-filling three-dimensional model of the crystalline structure of the Form D polymorph of CV-8972.

[0067] [Figure 2] FIG. 2 is a space-filling three-dimensional model of the crystalline structure of the Form D polymorph of CV-8972 at room temperature.

[0068] [Figure 3] FIG. 3 is a space-filling three-dimensional model of the crystalline structure of the Form A polymorph of CV-8972.

[0069] [Figure 4]FIG. 4 is an XRPD diffractogram of the starting material CV-8972.

[0070] [Figure 5] FIG. 5 shows the TGA and DSC thermograms of the starting material CV-8972.

[0071] [Figure 6] FIG. 6 shows the XRPD diffractograms of various forms of CV-8972.

[0072] [Figure 7] FIG. 7 is a polarized microscope image of the starting material CV-8972.

[0073] [Figure 8] FIG. 8 is a dynamic water vapor sorption isotherm plot.

[0074] [Figure 9] FIG. 9 shows the XRPD diffractograms of CV-8972 before and after dynamic water vapor sorption.

[0075] [Figure 10] FIG. 10 shows the XRPD diffractograms of CV-8972 in its anhydrous and rehydrated forms.

[0076] [Figure 11] FIG. 11 shows the XRPD diffractograms of various polymorphs of CV-8972.

[0077] [Figure 12] Figure 12 is a PLM image of a single crystal batch of C22H34Cl3N3O6 (CV-8972).

[0078] [Figure 13] Figure 13 shows a PLM image of the crystal used in the single crystal diffractometer.

[0079] [Figure 14]Figure 14 shows an image of a crystal mounted in a 100 micron Mitegen loop on a diffractometer.

[0080] [Figure 15] Figure 15 is an Ortep diagram of the asymmetric unit of the C22H34Cl3N3O6 crystal.

[0081] [Figure 16] Figure 16 shows one unit cell of a C22H34Cl3N3O6 crystal.

[0082] [Figure 17] Figure 17 is a diagram of the hydrogen bond network and pairing with counterions in a C22H34Cl3N3O6 crystal.

[0083] [Figure 18] FIG. 18 shows the calculated and measured XRPD diagrams of C22H34Cl3N3O6 crystal.

[0084] [Figure 19] FIG. 19 shows a PLM image of an anhydrous single crystal from recrystallized CV-8972.

[0085] [Figure 20] FIG. 20 is an image of an anhydrous single crystal from recrystallized CV-8972 mounted on the tip of a glass fiber.

[0086] [Figure 21] FIG. 21 is a thermal ellipsoid diagram of the asymmetric unit of a C22H32Cl3N3O5 crystal.

[0087] [Figure 22] Figure 22 shows one unit cell of a C22H32Cl3N3O5 crystal.

[0088] [Figure 23]Figure 23 is a diagram of the hydrogen bond network and pairing with counterions in a C22H32Cl3N3O5 crystal.

[0089] [Figure 24] FIG. 24 shows the calculated and measured XRPD diagrams of C22H34Cl3N3O6 crystal. DETAILED DESCRIPTION OF THE INVENTION

[0090] Detailed Description The recently identified compound CV-8972 shows promise as a therapeutic agent for treating a variety of conditions, including cardiovascular conditions, rheumatic diseases, fibrosis, and cancer. It has the IUPAC name 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate and the following structure: [ka] CV-8972, which has the structure: [ka] CV-8814 is cleaved into CV-8814 and nicotinic acid, which have the structure: CV-8814 has the structure: CV-8814 is converted to trimetazidine in the body over time. Both CV-8814 and trimetazidine inhibit the beta-oxidation of fatty acids, thus shifting mitochondrial metabolism towards oxidizing glucose, a more oxygen-efficient energy source. Nicotinic acid converts nicotinamide adenine dinucleotide (NAD) + It serves as a precursor for the synthesis of NAD. +CV-8972 promotes mitochondrial respiration and drives ATP synthesis, regardless of whether glucose or fatty acids are used as the carbon source. Thus, in vivo, the two sets of products resulting from the degradation of CV-8972 act synergistically to stimulate mitochondrial energy production in cardiac tissue and other cell types. 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.

[0091] U.S. Patent No. 10,556,013 also provides a scheme for the synthesis of CV-8972. This scheme involves the formation of the free base form of CV-8814 by reductive amination of 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol. Because it is difficult to isolate CV-8814 in solid form in this previous method, the product of this reaction is then converted to the hydrochloride salt of CV-8814. However, CV-8814 must be converted back to its free base form for use in the esterification reaction with nicotinic acid to produce CV-8972.

[0092] The present invention recognizes that crystalline CV-8972 exists in multiple polymorphic forms. One polymorph, Form A, is most stable under ambient temperature and relative humidity conditions and therefore has particular utility for the manufacture of pharmaceutical compositions. Due to the stability of Form A, compositions containing this polymorph can be easily stored and distributed without losing therapeutic efficacy. Accordingly, the present invention provides compositions containing crystalline CV-8972 polymorphs, methods of making such compositions, and methods of using them to treat various conditions in subjects.

[0093] Polymorphism of CV-8972 As described in the Examples below, crystalline CV-8972 can exist in at least five polymorphic forms: Form A, Form B, Form C, Form D, and Form E. Form A is a monohydrate, and Forms B, D, and E are anhydrates. Form C was not obtained in purified form, and therefore its hydration state could not be determined.

[0094] The crystals may be formed as a salt of CV-8972. For example, the crystals may be formed as the hydrochloride salt of CV-8972.

[0095] Figure 1 is a space-filling three-dimensional model of the crystal structure of the CV-8972 polymorph Form D. This polymorph is the trihydrochloride salt, with chloride ions shown in green.

[0096] Figure 2 is a space-filling three-dimensional model of the crystalline structure of the CV-8972 polymorph Form D at room temperature. This polymorph is the trihydrochloride salt, with chloride ions shown in green.

[0097] Figure 3 is a space-filling three-dimensional model of the crystal structure of the CV-8972 polymorph Form A. This polymorph is the trihydrochloride salt, with chloride ions shown in green.

[0098] Pharmaceutical Composition The present invention provides pharmaceutical compositions comprising crystalline polymorphs of CV-8972. For example, the compositions can contain crystalline CV-8972 of Form A, Form B, Form C, Form D, or Form E. The compositions can be substantially free of one or more other polymorphs. For example, the compositions can contain polymorph Form A and can be substantially free of polymorphs Form B, Form C, Form D, and Form E.

[0099] A composition containing a polymorph of CV-8972 may be substantially free of one or more other polymorphic forms of CV-8972 if the composition contains the predominant polymorph at a specified level of purity. Purity may be expressed as the amount of the predominant polymorph as a percentage of the total weight of two of the more polymorphic forms of CV-8972.

[0100] In certain embodiments, the total weight refers to the weight of all polymorphs of CV-8972 in the composition. For example, a composition containing polymorph Form A and substantially free of other polymorphs can contain Form A at a specified weight percentage of all polymorphs of CV-8972 in the composition. For example, the composition can contain Form A at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% by weight of all polymorphs of CV-8972 in the composition.

[0101] In certain embodiments, the total weight refers to the weight of the selected polymorph of CV-8972 in the composition. For example, a composition containing the Form A polymorph and substantially free of the Form B polymorph can contain Form A in a specified weight percentage of Forms A and B. For example, the composition can contain Form A in at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% by weight of Forms A and B of CV-8972 in the composition. Similarly, a composition containing the Form A polymorph and substantially free of Forms B and C polymorphs can contain Form A in a specified weight percentage of Forms A, B, and C. For example, the composition can contain at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% by weight of Form A of CV-8972 Forms A, B, and C in the composition.

[0102] Alternatively, or in addition, a composition containing a polymorph of CV-8972 may be substantially free of one or more other polymorphic forms of CV-8972, provided that the composition contains the minor polymorph at a level below a specified level. The presence of a minor polymorph may be defined as the amount of one or more minor polymorphs as a percentage of the total weight of the two more polymorphs of CV-8972.

[0103] In certain embodiments, the total weight is the weight of all polymorphs of CV-8972 in the composition. For example, a composition containing polymorph Form A and substantially free of other polymorphs may contain all polymorphs other than Form A at a specified 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%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1% by weight of all polymorphs of CV-8972 in the composition.

[0104] In certain embodiments, the total weight refers to the weight of the selected polymorph of CV-8972 in the composition. For example, a composition containing the Form A polymorph and substantially free of the Form B polymorph can contain Form B at a specified weight percentage of Forms A and B. For example, the composition can contain Form B at less than 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1% by weight of Forms A and B of CV-8972 in the composition. Similarly, a composition containing the Form A polymorph and substantially free of Forms B and C polymorphs can contain Forms B and C at a specified weight percentage of Forms A, B, and C. For example, the composition can contain Forms B and C at less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% by weight of Forms A, B, and C of CV-8972 in the composition.

[0105] The composition may include a polymorphic hydrochloride salt of CV-8972. The composition may include CV-8972 and chloride ions in a defined stoichiometric ratio. The composition may include CV-8972 and chloride ions in a 1:3 stoichiometric ratio.

[0106] The composition can include a hydrated form of CV-8972. The composition can include a monohydrate form of CV-8972, such as polymorph Form A. The composition can include an anhydrous form of CV-8972, such as polymorph Form B, Form D, or Form E.

[0107] The composition may be formulated for any route or mode of administration. The composition may be formulated for oral, cutaneous, intestinal, intraarterial, intramuscular, intraocular, intravenous, intranasal, oral, parenteral, pulmonary, rectal, subcutaneous, topical, or transdermal administration. The composition may be formulated for administration by injection or using or on an implantable medical device (e.g., a stent or drug-eluting stent or balloon equivalent).

[0108] The composition may be formulated as a single unit dose. The composition may be formulated as divided doses.

[0109] The composition may contain a prescribed dose of CV-8972. This dose is for administering CV-8972 in an amount of 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, or about It may contain 50mg to about 200mg, about 100mg to about 2000mg, about 100mg to about 1000mg, about 100mg to about 800mg, about 100mg to about 600mg, about 100mg to about 400mg, about 100mg to about 300mg, about 100mg to about 200mg, about 200mg to about 2000mg, about 200mg to about 1000mg, about 200mg to about 800mg, about 200mg to about 600mg, about 200mg to about 400mg, about 200mg to about 300mg, about 300mg to about 2000mg, about 300mg to about 1000mg, about 300mg to about 800mg, about 300mg to about 600mg, or about 300mg to about 400mg. This dose can contain about 10 mg, about 25 mg, about 50 mg, about 100 mg, about 200 mg, about 300 mg, or about 400 mg of CV-8972.

[0110] Pharmaceutical compositions containing CV-8972 polymorphs may be in a form suitable for oral use, such as tablets, troches, lozenges, fast-melt tablets, dispersible powders or granules, or capsules. Compositions for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents selected from sweeteners, flavoring agents, coloring agents, and preservatives to produce medicament-elegant and palatable preparations. Tablets contain the polymorphs in a mixture with non-toxic pharmaceutically acceptable additives. These additives can be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents, such as corn starch or alginic acid; binders, such as starch, gelatin, or acacia, 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 may also be provided as hard gelatin capsules, in which the compound is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin. The formulation may be encapsulated in an enteric coating to allow for controlled release of the CV-8972 polymorph in the gastrointestinal tract.

[0111] Dispersible powders and granules provide the compound in a mixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified, and for example, sweeteners, flavoring agents, and coloring agents may also be present.

[0112] The pharmaceutical composition may contain a mixture containing an erodible polymer, which promotes the swelling of the mixture in an aqueous environment. An erodible polymer is any polymer that degrades within the body within a physiologically relevant time frame. The erodible polymer may have other characteristics that promote the gradual release of the CV-8972 polymorph from the mixture. For example and without limitation, the polymer may be one or more of the following: biocompatible, i.e., not harmful to living tissue, hydrophilic, hygroscopic, or prone to forming a hydrogel.

[0113] Without wishing to be bound by theory, polymer-containing mixtures can promote gradual release through one or more mechanisms. For example, swelling of the mixture due to absorption of water can promote the diffusion of CV-8972 polymorphs from the mixture. Degradation of the polymer can also allow CV-8972 polymorphs to be released from the mixture. Osmotic pressure due to a high concentration gradient of the compound between the inside and outside of the mixture can also contribute to the diffusion of CV-8972 polymorphs from the mixture.

[0114] For example and without limitation, the polymer may be a cellulose derivative, a gelatin derivative, such as a cross-linked gelatin derivative, or a polyester derivative.

[0115] Cellulose derivatives include polymers of linear β(1→4)-linked D-glucose units with substitutions on one or more of the hydroxyl groups of each glucose unit. The substituents may be organic or inorganic and are usually attached via ester or ether linkages. Cellulose ester derivatives include carboxymethylcellulose (CMC), such as sodium carboxymethylcellulose, ethyl cellulose, ethylhydroxyethyl cellulose, ethylmethylcellulose, hydroxyethyl cellulose, hydroxyethylmethylcellulose, hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), and methylcellulose. 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.

[0116] A blend may contain multiple polymers or multiple polymer forms of the same polymer. For example, HPMC polymer forms may differ in various physical properties, including viscosity, degree of methoxy substitution, degree of hydroxypropoxyl substitution, or average molecular weight.

[0117] The viscosity of the HMPC polymer form can be determined by testing under standard conditions, including the concentration of HMPC in solution and the temperature of the solution. For example and without limitation, the HPMC concentration can be 1%, 1.5%, 2%, 2.5%, or 3%. For example and without limitation, the temperature of the solution 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.

[0118] Polymeric forms of cellulose derivatives such as HPMC can have a defined viscosity. For example and without limitation, the polymeric form of HPMC may have a viscosity of 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, 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.

[0119] The polymer form of cellulose derivatives such as HPMC can have different substitution degree of glucose unit.Substitution degree can be expressed as weight percentage of substitution or as molar ratio of substitution to glucose unit.For the cellulose derivatives such as HPMC that have two different substitutions, polymer form can be described by the substitution degree of each substitution.

[0120] Each polymer form of HPMC can independently have a defined degree of methoxy substitution. For example and without limitation, 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%.

[0121] Each polymer form of HPMC can independently have a defined degree of hydroxypropoxyl substitution. For example and without limitation, the degree of hydroxypropoxyl 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 about 11%, or about 9% to about 12%.

[0122] Each polymeric form of HPMC can independently have a defined average molecular weight, which 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.

[0123] When multiple forms of a polymer such as HPMC are present, one or more polymer forms can be present in a specified amount. For example and without limitation, a polymer such as HPMC can contain about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by weight of one polymer form.

[0124] The pharmaceutical composition may include a modified release formulation containing one or more polymorphs of CV-8972. The formulation contains a mixture comprising one or more polymorphs of CV-8972 and one or more erodible polymers, where the one or more erodible polymers promote the swelling of the mixture in an aqueous environment. Due to the hygroscopic and erodible properties of the polymer, the mixture can form a hydrogel that slowly degrades in the subject's digestive tract. Therefore, the mixture promotes the steady release of the polymorphs of CV-8972 and their metabolites into the blood circulation.

[0125] The mixture can contain a specified amount of CV-8972 polymorphs. The mixture can contain at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% by weight of CV-8972 polymorphs.

[0126] The mixture can contain the CV-8972 polymorph and the polymer in a specified weight ratio. For example and without limitation, the mixture can contain the CV-8972 polymorph and the polymer in a ratio 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 1:1 00 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, about 1:20 to about 10:1, about 1:20 to about 5:1, about 1:20 to about 2:1, about 1:10 to about 100:1, about 1:10 to about 50:1, about 1:10 to about 20:1, about 1:10 to about 10:1, about 1:10 to about 5:1, about 1:10 to about 2:1, about 1:5 to about 100:1, about 1:5 The amount of hydroxybenzoates may be about 1:3 to about 50:1, about 1:5 to about 20:1, about 1:5 to about 10:1, about 1:5 to about 5:1, about 1:5 to about 2:1, about 1:3 to about 100:1, about 1:3 to about 50:1, about 1:3 to about 20:1, about 1:3 to about 10:1, about 1:3 to about 5:1, or about 1:3 to about 2:1 by weight.

[0127] The pharmaceutical composition may be formulated for a particular route of administration. The medicament may be formulated for oral, enteral, intravenous, or rectal administration.

[0128] The pharmaceutical composition may be formulated as a unit dosage containing a specified amount of the CV-8972 polymorph. Unit dosages may range from about 5 mg, about 10 mg, about 20 mg, about 50 mg, about 100 mg, about 200 mg, about 500 mg, about 5 mg to about 10 mg, about 5 mg to about 20 mg, about 5 mg to about 50 mg, about 5 mg to about 100 mg, about 5 mg to about 200 mg, about 5 mg to about 500 mg, about 10 mg to about 20 mg, about 10 mg to about 50 mg, about 10 mg to about 100 mg, and about 10 mg to 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 of a polymorph of CV-8972.

[0129] The pharmaceutical composition may be formulated to produce a defined value for one or more parameters described below with respect to the methods of the invention. For example and without limitation, the parameters may be C max , administration and C max The interval between the achievement of T 1 / 2 Or it can be AUC.

[0130] The pharmaceutical composition of the present invention may contain excipients.For example and without limitation, the composition may contain sweeteners, flavoring agents, coloring agents or preservatives.The composition may contain one or more of mannitol, starch and magnesium stearate.

[0131] Administration of CV-8972 polymorphs to subjects The present invention provides a method for treating a condition in a subject by administering a polymorph of CV-8972. The polymorph can be Form A, Form B, Form C, Form D, or Form E. The polymorph of CV-8972 may be provided in a pharmaceutical composition as described above. In certain embodiments of the method, only Form A polymorph is provided.

[0132] The CV-8972 polymorph may be administered by any suitable route or mode of administration. For example, and without limitation, the CV-8972 polymorph may be administered orally, cutaneously, intestinally, intraarterially, intramuscularly, intraocularly, intravenously, intranasally, orally, parenterally, intrapulmonary, rectally, subcutaneously, topically, transdermally, by injection, or using or onto an implantable medical device (e.g., a stent or drug-eluting stent or balloon equivalent).

[0133] The polymorphs of CV-8972 may be provided according to a dosing regimen, which can include the amount of administration, the frequency of administration, or both.

[0134] Dosage can be administered at any suitable interval.For example and without limitation, dosage can be administered 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 2 days, once every 3 days, once every 4 days, once every 5 days, once every week, twice a week, three times a week, four times a week or five times a week.

[0135] The dose can contain a defined amount of CV-8972 that improves cardiac mitochondrial function, such as any of the doses described above for pharmaceutical compositions containing a polymorph of CV-8972.

[0136] The dose may be administered in a single dose, i.e., the dose may be administered as a single tablet, capsule, pill, etc. Alternatively, the dose may be administered in split doses, i.e., the dose may be administered as multiple tablets, capsules, pills, etc.

[0137] Administration may continue for a specified period of time. For example and without limitation, a dose may be administered for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks or longer.

[0138] The subject can be a human. The subject can be a human with a cardiovascular condition, a rheumatic condition, fibrosis, or cancer. The subject can be a human at risk for developing a cardiovascular condition, a rheumatic condition, fibrosis, or cancer. A subject can be at risk for developing a condition if they do not meet established criteria for a diagnosis of that condition but have one or more symptoms, markers, or other factors that indicate that they are likely to meet the diagnostic criteria for that condition in the future. The subject can be a pediatric, newborn, neonatologist, infant, child, adolescent, preteen, teenager, adult, or elderly subject. The subject can be in critical 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-hospital field setting.

[0139] Conditions that may be treated by polymorphs of CV-8972 The present invention provides methods of treating a condition in a subject by administering a polymorph of CV-8972. The condition can be any disease, disorder, or condition in which increased mitochondrial energy production provides a therapeutic benefit.

[0140] The condition may be a cardiac condition, for example, but not limited to, an aneurysm, angina, 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 arterial disease, refractory angina, rheumatic heart disease, stable angina, stroke, transient ischemic attack, unstable angina, or valvular heart disease.

[0141] Angina pectoris (angina) is chest pain or pressure, usually due to insufficient blood flow to the heart muscle. The pain or discomfort is located behind the sternum or on the left side and may radiate down the left arm, neck, jaw, or back. Several classifications of angina are known.

[0142] Stable angina, also known as exertional angina, is associated with myocardial ischemia. In stable angina, chest discomfort and related symptoms are usually triggered by some physical activity, such as running or walking, but the symptoms are minimized or disappear when the patient rests or takes sublingual nitroglycerin. Symptoms usually subside within a few minutes after activity and recur when activity is resumed. Symptoms can also be precipitated by cold weather, heavy meals, and emotional stress.

[0143] Unstable angina is angina that changes or worsens. Unstable angina has at least one of the following characteristics: (1) it occurs at rest or with minimal exertion and usually lasts more than 10 minutes; (2) it is severe and new, i.e., occurring within the past 4 to 6 weeks; and (3) it occurs in a crescendo pattern, i.e., it is significantly more severe, lasts longer, or occurs more frequently than before.

[0144] Cardiac syndrome X, also called microvascular angina, is characterized by angina-like chest pain in the setting of normal epicardial coronary arteries on angiography. Its primary cause is unknown, but a clearly contributing factor is endothelial dysfunction, which reduces flow in the cardiac's small resistance vessels. Microvascular angina may be part of the pathophysiology of ischemic heart disease.

[0145] Refractory angina is a chronic condition (≥3 months duration) in which angina (1) occurs in the setting of coronary artery disease (CAD), (2) cannot be controlled by a combination of optimal medical therapy, angioplasty, or bypass surgery, and (3) it is clinically established that reversible myocardial ischemia is the cause of the symptoms.

[0146] Administering polymorphs of CV-8972 can improve cardiac efficiency in subjects.Various definitions of cardiac efficiency exist in 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 cardiac output to the outside of the heart to the cardiac energy consumption by the left ventricle.See, for example, Lopaschuk G.D., et al., Myocardial Fatty Acid Metabolism in Health and Disease, Phys.Rev. 90:207-258(2010), which are incorporated herein by reference. Another definition is the ratio between stroke work and oxygen consumption, which in a normal human heart is in the range of 20-25%. Visser, F., Measuring cardiac efficiency: is it useful? Hear Metab. 39:3-4 (2008), incorporated herein by reference. Another definition is the ratio of stroke volume to mean arterial pressure. Any suitable definition of cardiac efficiency can be used to measure the effects of the compounds of the present invention.

[0147] CV-8972 polymorphs can be used to treat rheumatic disease, disorder or condition.As used herein, rheumatic disease, disorder or condition is any condition that affects joints, tendons, ligaments, bones, muscles or connective tissue, or is accompanied by pain in one or more of these tissues.Rheumatic disease, disorder or condition can mainly affect joints, tendons, ligaments, bones, muscles or connective tissue. Examples of such conditions include ankylosing spondylitis, autoimmune-related 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, Sjogren's syndrome, tendonitis, and vasculitis.

[0148] A rheumatic disease, disorder, or condition primarily affects the cardiovascular system and may have secondary effects on joints, tendons, ligaments, bones, muscles, or connective tissue. For example, and without limitation, the condition can be alcoholic cardiomyopathy, aneurysm, angina pectoris (including refractory angina and angina in the setting of heart failure), atherosclerosis, myocardial fibrosis, cardiomyopathy, cerebrovascular disease, claudication (e.g., peripheral claudication), congenital heart disease, coronary artery disease, coronary heart disease, cyanotic heart disease, diabetic cardiomyopathy, dilated cardiomyopathy, heart attack, heart failure, high blood pressure (hypertension), hypertrophic cardiomyopathy, ischemic cardiomyopathy, ischemic heart disease, left ventricular dysfunction, pericardial disease, peripheral arterial disease, rheumatic heart disease, stroke, transient ischemic attack, or valvular heart disease.

[0149] Rheumatic disease, disorder or condition may be rare muscle disease.For example and not limited to, this condition may be CAV3-associated distal myopathy, Duchenne muscular dystrophy, hypertrophic cardiomyopathy, isolated hyperCKemia, limb-girdle muscular dystrophy 1C, muscle myopathy, myositis or rippling muscle disease.Rare muscle disease may be associated with mutation in BICD2, CAV3 or DMD.

[0150] The rheumatic disease, disorder, or condition may be a glycogen storage disease. For example, and without limitation, the glycogen storage disease may be aldolase A deficiency, Andersen's disease, Cori's disease, Fanconi-Bickel syndrome, Haas's disease, Lafora's disease, McArdle's disease, Pompe's disease, Tarui's disease, or von Gierke's disease. The glycogen storage disease may be associated with a deficiency of an enzyme or protein, such as acid alpha-glucosidase, aldolase A, β-enolase, glucose transporter, glucose-6-phosphatase, glycogen branching enzyme, glycogen debranching enzyme, glycogen synthase, glycogenin-1, liver glycogen phosphorylase, muscle glycogen phosphorylase, muscle lactate dehydrogenase, muscle phosphofructokinase, muscle phosphoglycerate mutase, phosphoglycerate mutase, or phosphorylase kinase. Glycogen storage diseases can 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.

[0151] The rheumatic disease, disorder or condition may be another condition affecting the joints, tendons, ligaments, bones, muscles or connective tissue, such as acute kidney injury, cachexia, chemotherapy-induced nephropathy, contrast nephropathy, imbalance, weakness, pulmonary arterial hypertension, pulmonary fibrosis, sarcopenia, tinnitus or vertigo.

[0152] CV-8972 polymorphs may be used to treat fibrosis or diseases, disorders or conditions associated with fibrosis.In particular, this method is useful for treating diseases, disorders or conditions in which fibrosis in an organ or tissue is associated with a decrease in energy production by 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.

[0153] Fibrosis may be associated with, for example, secondary to, or lead to another condition. For example, and without limitation, fibrosis can include or be associated with adhesive capsulitis, aneurysm, angina pectoris, arteriosclerosis, arthrofibrosis, 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, endomyocardial fibrosis, glial scar, heart attack, heart failure, high blood pressure (hypertension), idiopathic pulmonary fibrosis, ischemic heart disease, keloid, mediastinal fibrosis, myelofibrosis, nephrogenic systemic fibrosis, old myocardial infarction, pericardial disease, peripheral arterial disease, Peyronie'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.

[0154] Polymorphs of CV-8972 may be used to treat cancer. For example and without limitation, the cancer may be bladder cancer, brain cancer, breast cancer, carcinoma, cervical cancer, colon cancer, colorectal cancer, gastric cancer, glioblastoma, glioma, head and neck cancer, kidney cancer, leukemia, liposarcoma, liver cancer, lung cancer, lymphoma, medulloblastoma, melanoma, muscle cancer, neuroblastoma, oligoastrocytoma, oligodendroglioma, osteosarcoma, ovarian cancer, pancreatic cancer, paraganglioma, prostate cancer, sarcoma, or thyroid cancer.

[0155] Synthesis of CV-8972 The present invention also provides a synthetic scheme for CV-8972 in which the free base form of CV-8814, formed as a product in a reductive amination reaction, can be used directly as a substrate in an esterification reaction. The present invention is based, in part, on the identification of conditions that improve the stability of the free base of CV-8814 and allow the free base form to crystallize. Thus, 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, the present invention provides a simpler, faster, and higher-yielding method for making CV-8972.

[0156] The present invention relates to a compound of formula (X): [ka] 1. A method for preparing 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol are reacted to give a compound of formula (IX): [ka] generating the free base form of reacting the free base form of the compound of formula (IX) with nicotinic acid to produce a compound of formula (X). This is done by carrying out The present method provides a method that does not involve producing a salt form of the compound of formula (IX). 2,3,4-Trimethoxybenzaldehyde has the following structure: [ka] It has. 2-(piperazin-1-yl)ethan-1-ol has the following structure: [ka] It has.

[0157] Nicotinic acid has the following structure: [ka] It has.

[0158] The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol can include one or more of a solvent, a catalyst, or other chemicals. The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol can include one or more of sodium triacetoxyborohydride, acetic acid, and 2-methyltetrahydrofuran.

[0159] The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol can be carried out at a specified temperature, such as about 10°C to about 30°C, about 15°C to about 30°C, about 20°C to about 30°C, about 25°C to about 30°C, about 10°C to about 25°C, about 15°C to about 25°C, about 20°C to about 25°C, about 10°C to about 20°C, or about 15°C to about 20°C.

[0160] The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol may be free of a particular solvent, catalyst, or other chemical. The step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol may be free of dichloromethane.

[0161] Reacting the free base form of the compound of formula (IX) with nicotinic acid can produce the free base form of the compound of formula (X).

[0162] The step of reacting the free base form of the compound of formula (IX) with nicotinic acid can include one or more of a solvent, a catalyst, or other chemicals. The step of reacting the free base form of the compound of formula (IX) with nicotinic acid can include one or more of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, 4-(dimethylamino)pyridine, and dichloromethane.

[0163] The step of reacting the free base form of the compound of formula (IX) with nicotinic acid can be carried out at a temperature of 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.

[0164] The method can include converting the free base form of the compound of formula (X) into a salt form of the compound of formula (X). The salt form of the compound of formula (X) can be an HCl salt. The salt form of the compound of formula (X) can be a monohydrate.

[0165] The step of converting the free base form of the compound of formula (X) to a salt form of the compound of formula (X) can include one or more of a solvent, a catalyst, or other chemical. The step of converting the free base form of the compound of formula (X) to a salt form of the compound of formula (X) can include one or more of HCl and methyl ethyl ketone.

[0166] The step of converting the free base form of the compound of Formula (X) to the salt form of the compound of Formula (X) can be carried out at a specified temperature. The step of converting the free base form of the compound of Formula (X) to the salt form of the compound of Formula (X) can be carried out at about 40°C to about 60°C, about 45°C to about 60°C, about 50°C to about 60°C, about 55°C to about 60°C, about 40°C to about 55°C, about 45°C to about 55°C, about 50°C to about 55°C, about 40°C to about 50°C, about 45°C to about 50°C, about 40°C to about 50°C, about 40°C, about 45°C, about 50°C, about 55°C, or about 60°C.

[0167] 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 anhydrates. The method can include converting the compound of formula (X) from a first crystalline form to a second crystalline form. The first and second crystalline forms can each independently be Form A, Form B, Form C, Form D, or Form E. The method can include one or more of converting the compound of formula (X) from an anhydrous form to a hydrated form, from a hydrated form to an anhydrous form, from one anhydrous form to another anhydrous form, and from one hydrated form to another hydrated form.

[0168] The step of converting the salt form of the compound of Formula (X) from a first crystalline form to a second crystalline form may include one or more of the steps of changing the solvent of the salt form of the compound of Formula (X) and incubating the salt form of the compound of Formula (X) at about 60°C.

[0169] The process may be carried out without the use of one or more of a solvent, catalyst or other chemicals. The process may be carried out without the use of one or more of dioxane, ethyl acetate or potassium carbonate.

[0170] The method may include purifying the free base form of the compound of formula (IX).The method may include crystallizing the free base form of the compound of formula (IX). [Example]

[0171] Example 1 summary A comprehensive polymorph screening of CV-8972, having the structure of Formula (X), was undertaken. The starting material, CV-8972, was characterized by X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), dynamic vapor sorption (DVS), and polarized light microscopy (PLM). Data indicated that this material was crystalline in nature and had an XRPD pattern similar to that of Form A. Starting with Form A, polymorph / single crystal screening experiments were set up under 34 conditions using vapor diffusion, slow solvent evaporation, and cooling crystallization methods. Five unique XRPD patterns were observed, including Form A, Form B, Form A+C, Form D, and Form E. Form A is a monohydrate form, confirmed by a single crystal structure. Form D is anhydrous and was also confirmed by a single crystal structure. Form E is an anhydrous form produced by dehydration of Form A at approximately 90°C. Form B is the known anhydrate from a separate study. Form C was not obtained in pure form during this study, but rather appeared as a mixture of Forms A and C. Water activity analysis showed that Form E converted to Form A under all conditions tested. Furthermore, Form E was shown to partially convert to Form A upon exposure to ambient temperature and humidity. Furthermore, results due to slurry competition between both anhydrous Forms D and E also showed that both forms converted to Form A during this experiment. These results suggest that Form A is the most stable form at ambient temperature and humidity.

[0172] Characterization of Form A The CV-8972 starting material was characterized using X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and polarized optical microscopy (PLM).

[0173] Figure 4 shows the XRPD diffractogram of the starting material CV-8972. The XRPD results indicated that the starting material was highly crystalline. Comparison of the XRPD of the starting material with a previously known polymorph indicated that it was Form A.

[0174] Figure 5 shows the TGA and DSC thermograms of the starting material CV-8972. The TGA thermogram is shown in green, and the DSC thermogram is shown in blue. As shown by the TGA and DSC data, a weight loss of approximately 3.46% was observed up to 150°C before decomposition. The DSC showed a slight endotherm (peak) at 85.3°C and a likely melting endotherm (onset) at 214.6°C, followed by decomposition and a melting point of 131.7°C (peak).

[0175] Figure 6 shows XRPD diffractograms of various forms of CV-8972. Starting CV-8972 is shown in blue, CV-8972 after 8 hours of incubation at 90°C is shown in red, and CV-8972 after 2 hours of incubation at 65°C in vacuum is shown in purple. To determine whether the slight DSC endotherm at 85.3°C corresponds to a polymorphic phase transition or dehydration, XRPD was performed on Form A after storing CV-8972 in an oven at 90°C for 8 hours. The data showed that Form A converted to Form E.

[0176] Figure 7 is a polarized light microscope image of the starting material CV-8972. The crystals were observed by PLM to have a very plate-like "mica-like" morphology.

[0177] Figure 8 is a dynamic water vapor sorption isotherm plot. Sorption cycles are shown in red, cycle 1 desorption in blue, and cycle 2 sorption in green. DVS results showed that the water uptake of CV-8972 was <0.2% at 25°C and 80% relative humidity (RH), indicating that the starting material was non-hygroscopic. However, above 80% RH, the mass change increased significantly, indicating possible deliquescence.

[0178] Figure 9 shows the XRPD diffractograms of CV-8972 before and after dynamic water vapor sorption. The pre-DVS data is shown in red, and the post-DVS data is shown in blue. The XRPD of the post-DVS sample showed weak crystalline peaks but was mostly similar to the starting material.

[0179] Figure 10 shows XRPD diffractograms of CV-8972 in its anhydrous and rehydrated forms. Data from the starting material is shown in blue, data after incubation in a vacuum oven for 2 hours is shown in red, and data from the heated material exposed to ambient relative humidity is shown in green. To monitor Form A in its anhydrous state, it was placed in a vacuum oven at 65°C for 2 hours before XRPD analysis. The XRPD results indicated that this process produced a new anhydrous form of this material, assigned as Form E. Rehydration of Form E when exposed to ambient RH resulted in partial conversion to Form A.

[0180] X-ray powder diffraction data are difficult to interpret due to highly preferential orientation, which results in large variations in peak intensities from one sample preparation to the next. To minimize this effect, single crystal X-ray diffraction was used to obtain the crystallographic structure and calculate the X-ray powder diffraction that would be observed in an ideal sample in which there is no preferred orientation.

[0181] Polymorph / Single Crystal Screening Starting with Form A, polymorph screening experiments were set up under 34 conditions using the following methods: slurry conversion, liquid vapor diffusion, slow solvent evaporation, and slow cooling. The solubility of the starting material was estimated at room temperature (RT). An accurately weighed sample weighing approximately 2 mg was added to a 3 mL glass vial. Solvent was then added to the vial in stepwise fashion (50 / 50 / 200 / 700 μL) until the solid dissolved or a total volume of 1 mL was reached. The solubility of the starting material in various solvents is shown in Table 1. [Table 1-1] [Table 1-2]

[0182] Results from solubility analysis were used to guide solvent selection in polymorph screening. Polymorph screening experiments were performed using various crystallization or solid-state transition methods. The polymorph screening experiments are summarized in Table 2. [Table 2]

[0183] Figure 11 shows XRPD diffractograms of various polymorphs of CV-8972, with Form A shown in blue, Form B shown in green, a mixture of Forms A and C shown in navy blue, Form D shown in orange, and Form E shown in purple.

[0184] The various crystalline forms of CV-8972 are summarized in Table 3. [Table 3] *Single crystal structures of these forms are available and will be presented in separate reports. # Information obtained from another report

[0185] Liquid vapor diffusion experiments were performed under various solvent conditions. Approximately 15–25 mg of starting material was dissolved in an appropriate solvent in a 3 mL vial to obtain a clear solution. This solution was then placed in a 20 mL vial containing 3 mL of a volatile solvent. The 20 mL vial was sealed with a cap and kept at room temperature to allow sufficient time for the organic vapor to interact with the solution. The precipitate was isolated for XRPD analysis. Table 4 summarizes the results of the liquid vapor diffusion experiments. [Table 4-1] [Table 4-2]

[0186] Slow solvent evaporation experiments were performed under various conditions. Briefly, saturated solutions of starting materials prepared in various solvents were added to HPLC vials. The visually clear solutions were covered with Parafilm® containing 5-10 pinholes and subjected to solvent evaporation at room temperature. Solids were isolated for XRPD analysis. Table 5 summarizes the results from the slow solvent evaporation experiments. [Table 5]

[0187] Slow cooling experiments were performed with two different solvent systems. Approximately 10-15 mg of starting material was suspended in the appropriate solvent in a 2 mL glass vial at room temperature. The suspension was then heated to 50°C, equilibrated for approximately 2 hours, and filtered using a nylon membrane (pore size 0.22 μm). Each filtrate was slowly cooled to 5°C at a rate of 0.1°C / min. Table 6 summarizes the results from the slow cooling experiments. [Table 6]

[0188] Slurry conversion experiments were carried out at room temperature in various solvent systems. Approximately 20 mg of starting material was suspended in 0.1 mL of solvent in an HPLC vial. The suspension was magnetically stirred at room temperature for 48 hours, after which the residual solid was isolated for XRPD analysis. Table 7 summarizes the results from the slurry conversion experiments. [Table 7]

[0189] conclusion Form A was successfully characterized to understand its behavior. A comprehensive polymorph screening under 34 different conditions was performed. Five polymorphs of CV-8972 were identified during screening, including Form A, Form B, a mixture of Forms A and C, Form D, and Form E. Forms D and E are anhydrous, Form A is a monohydrate, and Form B is a hydrate with unknown stoichiometry. The phase origin of Form C is unknown because it was not obtained in pure form. It always crystallized as a mixture with Form A. Based on the polymorph screening, it is clear that CV-8972 has a tendency to form multiple polymorphs. The current study concluded that Form A is the best form for development of CV-8972 and is a stable monohydrate form, as well as the most stable form under ambient temperature and humidity conditions.

[0190] Apparatus and Method Form A was analyzed and used as starting material to screen for other polymorphs.

[0191] XRPD was performed using a Panalytical X'Pert3 Powder XRPD in a Si zero background holder. 2θ positions were calibrated against a Panalytical Si reference standard disc. The instrument parameters used for XPRD are listed in Table 8. [Table 8]

[0192] TGA data was collected using a TA Discovery550 TGA manufactured by TA Instruments. DSC was performed using a TA Q2000 DSC manufactured by TA Instruments. The DSC was calibrated with an indium reference standard and the TGA was calibrated using a nickel reference standard. The detailed parameters used for TGA and DSC are listed in Table 9. [Table 9]

[0193] Polarized light microscopy (PLM) photographs were captured on a Nikon DS-Fi2 upright microscope at room temperature. Low viscosity microscope immersion oil (Resolve®) was used to disperse the powder crystals.

[0194] Example 2 summary To determine the crystal structure of CV-8972, monohydrate single crystals were grown and suitable single crystals were used to collect whole crystal X-ray diffraction (SCXRD) data at 199 K. A crystal structure was obtained with an R value of 0.0303 (I > 2σ(I)). The structure indicated that this crystalline form was the monohydrate tri-HCl salt.

[0195] Crystal growth and SCXRD preparation By slow cooling, C 22 H 34 Single crystals of Cl3N3O6 (CV-8972) were obtained by weighing 163.2 mg of starting material into a 2 mL glass vial, adding 0.100 mL of water to dissolve the solid at 50 °C, and then slowly cooling the solution to 10 °C over 12 hours before harvesting.

[0196] Figure 12 shows the C 22 H 34 PLM image of a single crystal batch of Cl3N3O6 (CV-8972). The bar represents 100 μm.

[0197] Figure 13 shows a PLM image of the crystal used in the single crystal diffractometer. The bar represents 100 μm. Thick needles were selected and trimmed to a uniform block size of 200 × 160 × 100 μm. The sample was mounted in a 100 mm MiTeGen MicroLoop™ using low viscosity cryo-oil (MiTeGen LV CryoOil™).

[0198] Figure 14 shows an image of a crystal mounted in a 100 micron Mitegen loop on a diffractometer.

[0199] Single crystal structure determination A total of 9576 frames were collected using Bruker Apex3 v2018-7.2. The total exposure time was 18 hours (exposure time was adjusted based on 2θ). Frames were integrated with the Bruker SAINT software package using a narrow frame algorithm. Integration of the data using an orthorhombic unit cell yielded a total of 157237 reflections, of which 5641 were independent (average multiplicity 27.874, completeness = 99.8%, R int =4.41%, R sig =1.34%), 5388 (95.51%) were within the 2σ(F 2 a = 7.8826(2) Å, b = 12.4776(3) Å, c = 52.3580(13) Å, volume = 5149.7(2) Å 3 The final cell constant is based on the refinement of the XYZ centroid of 1406 reflections over 20σ(I) with 11.75°<2θ<100.6°. The data were corrected for absorption effects using a multiscan method (SADABS). The minimum-to-maximum ratio of the apparent transmittance was 0.788. The calculated minimum and maximum values ​​of the transmittance coefficient (based on crystallite size) are 0.5340 and 0.7160.

[0200] Solving this structure, formula unit C 22 H 34 For Cl3N3O6, refinement was performed using the SHELXTL software package integrated into Olex2, with Z=8 and using the orthorhombic space group Pbca. One asymmetric unit contains one whole API molecule. F using 330 variables (0 restraints) 2The final anisotropic full-matrix least-squares refinement based on converged with R1 = 3.03% for the observed data and wR2 = 7.98% for all data. The goodness of fit was 1.041. The maximum peak in the final difference electron density synthesis was 0.358 e- / Å3 (0.81 Å from Cl1) and the maximum hole was -0.438 e- / Å3 (0.66 Å from Cl1). The hydrogen positions and most of the thermal ellipsoids were treated as riding models (using AFIX23, AFIX43, and AFIX137). However, key hydrogen atoms involved in hydrogen bonding and salt formation were freely refined without any constraints. Based on the final model, the calculated density was 1.400 g / cm. 3 and F(000), 2288e-. Table 10 shows the C 22 H 34 The crystallographic parameters of Cl3N3O6 crystal are summarized below. [Table 10-1] [Table 10-2]

[0201] Figure 15 shows the C 22 H 34 Ortep diagram of the asymmetric unit of a Cl3N3O6 crystal. 22 H 34 An Ortep diagram of the asymmetric unit of the Cl3N3O6 crystal demonstrates that the API is a monohydrate tri-HCl salt, as a 1:3:1 (API:HCl:H2O) ratio was observed.

[0202] Figure 16 shows the C 22 H 34 A unit cell of a Cl3N3O6 crystal is shown.

[0203] Figure 17 shows the C 22 H 34Diagram of the hydrogen bond network and counterion pairing in Cl3N3O6 crystal. The diagram shows that three hydrochloride molecules are deprotonated, while three nitrogens are protonated. A water molecule acts as a hydrogen bond donor to bridge two chloride anions. Table 11 shows the C 22 H 34 We summarize crystallographic measurements of hydrogen bonds and counterion pairs in Cl3N3O6 crystals. [Table 11]

[0204] Symmetry transformations used to generate equivalent atoms: #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

[0205] The final cif file was reviewed using Platon using Olex2 locally, which revealed only one level C alert (three reflections missing) along with eight level G alerts. To prevent this issue, an extensive data collection strategy was implemented, and the completeness and multiplicity of this dataset were 99.8% and 27.87, respectively.

[0206] Figure 18 shows C 22 H 34 Calculated and measured XRPD diagrams for Cl3N3O6 crystals are shown. The calculated XRPD diffractogram is shown in red, and the measured XRPD diffractogram is shown in blue. Powder X-ray diffraction was obtained for this batch and compared to the calculated pattern based on this crystal structure using Mercury. The experimental peak positions and intensities are in good agreement with the calculated pattern. Apparatus and Method

[0207] X-ray intensity data were measured on a Bruker Venture X-ray diffractometer at 199.0 K (controlled by an Oxford Cryostream 800). Monochromatic Cu Kα radiation (λ = 1.54178 Å, voltage = 50 kV, current = 1.1 mA) from an Incoatec microfocus source (1 μS 3.0) was used as the x-ray source. Intensity data were collected by a Photon II detector.

[0208] Polarized light micrographs were captured on a Nikon DS-Fi2 upright microscope at room temperature.

[0209] XRPD was performed using a Panalytical X'Pert3 Powder XRPD in a Si zero background holder. 2θ positions were calibrated against a Panalytical Si reference standard disc.

[0210] Example 3 summary To determine the crystal structure of CV-8972, an anhydrous single crystal of CV-8972 was grown at 102 K, and a suitable single crystal was used for full SCXRD data collection. A crystal structure was obtained with an R value of 0.0328 (I > 2σ(I)). The structure indicated that this crystalline form was the anhydrous tri-HCl salt.

[0211] Crystal growth and SCXRD preparation Single crystals of the anhydrous triHCl salt of CV-8972 were obtained by liquid vapor diffusion of MTBE in MeOH solution. Briefly, a saturated solution of CV-8972 in MeOH was obtained at room temperature and filled into a 2 mL glass vial, which was then placed inside a larger 20 mL vial containing 2 mL of MTBE. When the vial showed the presence of white crystalline material, the vial was removed.

[0212] FIG. 19 shows a PLM image of an anhydrous single crystal from recrystallized CV-8972.

[0213] Figure 20 shows an image of an anhydrous single crystal from recrystallized CV-8972 mounted on the tip of a glass fiber. The colorless crystal was then placed on the SCXRD instrument.

[0214] Single crystal structure determination A colorless crystal was mounted on the tip of a glass fiber. X-ray intensity data were measured at a temperature of 102 K using the omega / phi scan technique on a Bruker D8 Quest PHOTON 100 CMOS X-ray diffractometer system equipped with monochromated Mo Kα radiation (λ=0.71073 Å, sealed tube) from an Incoatec microfocus source (IμS). Data were collected in 1660 frames with a 10 second exposure time. Crystallographic data: C 22 H 32 O5N3Cl3: a=6.9940(6)Å, b=10.5742(9)Å, c=17.5786(14)Å, α=78.252(2)°, β=82.823(2)°, γ=82.476(2)°, V=1255.37(18)Å 3 , Z=2, FW=524.85, μ=0.403mm-1, d=1.389g / cm 3 , F(000)=552.

[0215] The crystallographic data and structure refinement for j1_a are presented in Table 12. [Table 12-1] [Table 12-2]

[0216] Atomic coordinates for j1_a (×10 4 ) and the equivalent isotropic displacement parameter (Å 2 x10 3 ) are presented in Table 13. Define U(eq) as one-third of the trace of the orthogonal Uij tensor. [Table 13-1] [Table 13-2]

[0217] The bond lengths [Å] for j1_a are presented in Table 14. [Table 14-1] [Table 14-2]

[0218] The angles [degrees] for j1_a are given in Table 15. [Table 15-1] [Table 15-2] [Table 15-3]

[0219] Of 8732 unique reflections collected up to a maximum theta angle of 32.02° (resolution 0.67 Å), 7553 were observed (I > 2σ(I)). The linear extinction coefficient for Mo Kα radiation is 0.403 mm -1 The data were integrated using the manufacturer's SAINT software and corrected for absorption effects using the multiscan method (SADABS).

[0220] Subsequent solution and refinement were performed using the SHELXTL-2014 solution package running on a Pentium® computer. The structure was solved by direct methods using the SHELXTL-2014 software package. Non-hydrogen atom scattering factors were adopted from literature tables. Non-hydrogen atoms were located from successive difference Fourier map calculations. In the final cycle of each refinement, all non-hydrogen atoms were refined with anisotropic displacement parameters. Except for H(1), H(2), and H(3) on the N(1), N(2), and N(3) atoms of the molecule, which were located from the difference Fourier maps and refined with appropriate restraints, the C-H bond lengths were assumed to be mÅ (m = 0.990 for CH2 groups, m = 0.980 for CH3 groups, and m = 0.950 for Ph-H groups), and the remaining hydrogen atom positions were calculated to rest on the carbons to which they were attached. The temperature factors of hydrogen atoms were fixed at n times the isotropic temperature factors of the C atoms to which they are bonded (n = 1.2 for CH2, Ph-H groups, and n = 1.5 for CH3). The crystal system of 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(I)) are R1 = 0.0328, wR2 = 0.0926, and the values ​​for all unique reflections are R1 = 0.0408, wR2 = 0.0975. The goodness of fit index for all data is 1.014. 0.549 to -0.459 e / Å 3 Peaks in the final difference map, ranging from , to , are chemically insignificant. We are working to resolve as many alerts generated by the CheckCIF program as possible. The current highest alert is at level G.

[0221] Figure 21 shows the C 22 H 32 1 is a thermal ellipsoid diagram of the asymmetric unit of the ClN0 crystal, demonstrating that this form is the anhydrous tri-HCl salt form.

[0222] Figure 22 shows C 22 H 32 A unit cell of a Cl3N3O5 crystal is shown.

[0223] Figure 23 shows C 22 H 32 Diagram of the hydrogen bond network and counterion pairs in Cl3N3O5 crystal.

[0224] Figure 24 shows C 22 H 34 Calculated and measured XRPD diagrams of crystalline Cl3N3O6 are shown. The calculated XRPD diffractogram is shown in red and the measured XRPD diffractogram is shown in green.

[0225] The compound crystallizes in the triclinic space group P-1 (number 2). The asymmetric unit has the formula C 22 H 32 The molecule contains one cation / anion salt (anhydrous triHCl salt) with 05N3Cl3. There may be some intramolecular H-bonds between N(1)-H(1)...Cl(3) (with a distance of 2.9634 (10)), N(2)-H(2)...Cl(2) (with a distance of 2.9822 (9)), and N(3)-H(3)...Cl(1) (with a distance of 3.0120 (9)). The structure solution, refinement, and calculation of derived results were performed using the SHELXTL-2014 computer program package. Neutral atom scattering factors were Cromer and Waber factors, and real and imaginary anomalous dispersion corrections were Cromer factors.

[0226] Example 4 introduction CV-8972 was synthesized according to Scheme 1. Scheme 1: [ka] [ka] [ka]

[0227] Step 1 is a reductive amination using the starting materials 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol using sodium triacetoxyborohydride (STAB) as the reducing agent in the presence of catalytic acetic acid (AcOH) and 2-methyltetrahydrofuran (2-MeTHF) as the solvent. After the reaction is complete, aqueous workup, solvent switching to MTBE, and recrystallization from MTBE / n-heptane forms the intermediate CV-8814 free base (CV8814 free base).

[0228] In step 2, CV-8814 free base (CV8814 free base) undergoes acid coupling with nicotinic acid in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and catalytic 4-(dimethylamino)pyridine (DMAP) in dichloromethane (DCM) solvent. After aqueous workup, CV8972 free base is formed. A solvent exchange to 2-butanone (MEK) followed by addition of concentrated HCl in MEK forms the CV8972 monohydrate intermediate.

[0229] The final step 3 is form conversion in a mixture of water, methanol and MEK at 60°C ± 5°C, followed by precipitation by addition of MEK to obtain the desired Form A of the final product CV-8972 by XRPD analysis.

[0230] Manufacturing Details Manufacturing details are provided in Table 16. [Table 16]

[0231] Generation Details Step 1, formation of CV8814 free base (2493-1903-00487), was carried out according to Scheme 2. Scheme 2: [ka]

[0232] Production details for step 1 are provided in Table 17. [Table 17-1] [Table 17-2]

[0233] 1) Sodium triacetoxyborohydride (STAB; 60.3 kg; CHP Lot Number: 181-190220) and 2-MeTHF (189.2 kg; CHP Lot Number: 234-190227) were charged to reactor R-401. 2) The contents of R-401 were stirred and the temperature was adjusted to 15°C ± 5°C. 3) 2,3,4-Trimethoxybenzaldehyde (27.5 kg; CHP Lot Numbers: 275-190311, 142-190212) and 2-MeTHF (71.1 kg; CHP Lot Number: 234-190227) were charged to reactor R-402. 4) Agitation in reactor R-402 was started. 5) 2-(piperazin-1-yl)ethan-1-ol (20.0 kg; CHP lot number: 173-190220) and 2-MeTHF (47.3 kg; CHP lot number: 234-190227) were charged to reactor R-402. 6) The contents of reactor R-402 were stirred for at least 5 minutes. 7) Acetic acid (1.41 kg: CHP Lot No.: 151-190214) was charged to reactor R-401 while maintaining the temperature of the mixture below 25°C. 8) T max =14.4℃ 9) The contents of Reactor R-402 were transferred over 1 hour and 19 minutes while maintaining the temperature of the mixture below 25°C. 10) T max =29.2°C (temperature exceeded range during addition). 11) It was considered good to move the reaction forward. 12) The temperature of the contents of R-401 was adjusted to 20°C ± 5°C and stirred at 20°C ± 5°C for at least 6 hours. 13) After approximately 19 hours, the contents of R-401 were sampled. 14) Analysis of the sample by QC showed that no peaks for 2,3,4-trimethoxybenzaldehyde were detected (specification ≤ 1.5 area%). 15) The temperature of the contents of R-401 was adjusted to 15°C ± 5°C. 16) Water (247.5 kg; CHP Lot No.: 232-190227) was charged to R-401 while maintaining the temperature below 25°C. 17)T max =14.6℃ 18) The temperature of the contents of R-401 was adjusted to 20°C ± 5°C and the contents were stirred for 30 minutes. 19) The contents of R-401 were allowed to stand for 30 minutes. 20) The phases were separated (cut) by transferring the aqueous layer to R-402. 21) MTBE (203.8 kg; CHP lot number: 207-190222) was charged into R-402. 22) The temperature of the contents of R-402 was adjusted to 0°C ± 5°C. 23) 50% NaOH (68.3 kg; CHP Lot Numbers: 145-190213, 150-190214) was charged to the R-402 while maintaining the temperature below 25°C. 24)T max =11.5℃ 25) After the addition was complete, the temperature of the R-402 contents was adjusted to 20°C ± 5°C. 26) The contents in the R-402 were stirred for at least 30 minutes, and then the contents were allowed to settle for at least 30 minutes. 27) The aqueous layer was transferred to reactor R-401, and the organic layer was left in reactor R-402, allowing for phase separation. 28) MTBE (61.0 kg; CHP lot number: 207-190222) was charged into R-401. 29) The temperature of the contents of R-401 was adjusted to 20°C ± 5°C. 30) The contents in R-401 were stirred for at least 15 minutes, and then the contents were allowed to settle for at least 15 minutes. 31) The aqueous layer was transferred to a drum and the organic layer was left in reactor R-401, allowing for phase separation. 32) The pH of the FIO was confirmed to be 13.20 in the aqueous layer of the drum. 33) The contents of R-402 were transferred to R-401. 34) 20% NaCl solution (94.4 kg; NaCl: 19.0 kg, CHP Lot No.: 156-190214; water (75.9 kg; CHP Lot No.: 232-190227) was charged to R-401, stirred for at least 15 minutes, and allowed to stand for at least 30 minutes. 35) The aqueous layer was transferred to a drum and phase separation was carried out. 36) The solution in R-401 was distilled under reduced pressure, maintaining a temperature of <45°C, to a total volume of approximately 55 L. 37) MTBE (61.1 kg; CHP lot number: 207-190222) was charged into R-401. 38) The contents in R-401 were distilled under reduced pressure, maintaining a temperature of <45°C, to a total volume of approximately 82 L. 39) MTBE (61.1 kg; CHP lot number: 207-190222) was charged into R-401. 40) The contents in R-101 were distilled under reduced pressure, maintaining a temperature of <45°C, to a total volume of approximately 82 L. 41) The contents of R-401 were sampled (IPC sample: 2493-1903-00484-85-01) and the water content of the solution was confirmed by KF analysis. 42)KF=1.6% (standard ≦0.5%) 43) MTBE (61.1 kg; CHP lot number: 207-190222) was charged into R-401. 44) The contents in R-101 were distilled under reduced pressure, maintaining a temperature of <45°C, to a total volume of approximately 82 L. 45) The contents of R-401 were sampled (IPC sample: 2493-1903-00484-88-01) and the water content of the solution was confirmed by KF analysis. 46)KF=0.8% (standard ≦0.5%) 47) MTBE (61.1 kg; CHP lot number: 207-190222) was charged into R-401. 48) The contents in R-101 were distilled under reduced pressure, maintaining a temperature of <45°C, to a total volume of approximately 82 L. 49) The contents of R-401 were sampled (IPC sample: 2493-1903-00484-91-01) and the water content of the solution was confirmed by KF analysis. 50)KF=0.4%(Standard≦0.5%) 51) MTBE (30.9 kg; CHP lot number: 207-190222) was charged into R-401. 52) The temperature of the contents of R-401 was adjusted to 40°C ± 5°C. 53) Heptane (56.3 kg; CHP Lot No.: 233-190227) was charged to R-401 over 8 minutes while maintaining the temperature at 40°C ± 5°C. 54)T min =38.1℃ 55) An FIO sample (FIO Sample: 2493-1903-00484-100-01) was taken to monitor the MTBE:heptane ratio of the contents in R-401. The ratio was 1.5:6.0. 56) The temperature of the contents of R-401 was adjusted to 28°C ± 5°C (target 26°C - 29°C) over at least 30 minutes and stirred at this temperature for at least 30 minutes. 57) The formation of a solid was observed. 58) The temperature of the contents of R-401 was adjusted to 30°C ± 3°C and stirred for at least 30 minutes. 59) Heptane (56.4 kg; CHP Lot No.: 233-190227) was charged to R-401 over 24 minutes while maintaining the temperature at 30°C ± 5°C. 60)T min =30.2℃. 61) The temperature of the contents of R-401 was adjusted to 30°C ± 3°C and stirred for at least 20 minutes. 62) The temperature of the contents of R-401 was adjusted to 20°C ± 5°C over at least 30 minutes and stirred for at least 20 minutes. 63) The temperature of the contents of R-401 was adjusted to 5°C ± 5°C over at least 30 minutes and stirred for at least 30 minutes. 64) The solid was collected on a filter FD-400. 65) The contents of Filter-FD-400 were washed with cold heptane (49.4 kg; CHP Lot No.: 233-190227). 66) The contents of Filter-FD-400 were dried under vacuum at ≦25° C. with a N 2 flow for at least 16 hours. 67) An IPC sample (IPC sample: 2493-1903-00484-122-01) was submitted to QC for LOD. · LOD=0.20% (standard ≦0.5%) 68) The product CV-8814 free base (CV8814 free base) was double wrapped, formed into an S-shape, and weighed. 69) Analysis of Dry Matter (IPC Sample: 2493-1903-00484-122-01): Appearance: White to off-white solid Weight 34.0 kg (78.0% yield) · HPLC purity=100.0% · 1H NMR: Consistent with structure. 70) A 5 kg portion of CV-8814 free base (CV8814 free base) was removed from the bulk material, double wrapped, configured into an S-shape, and reserved for release under lot number 2493-1903-00484.

[0234] Step 2, the formation of CV8972 monohydrate (2479-1903-00489), was carried out according to Scheme 3. Scheme 3: [ka]

[0235] Production details for steps 2a and 2b are provided in Table 18. [Table 18-1] [Table 18-2]

[0236] 1) Nicotinic acid (17.1 kg, CHP Lot No. 201-190222) and DCM (153.0 kg, CHP Lot No. 328-190326) were charged to reactor R-401. 2) The contents of R-401 were stirred and the temperature was adjusted to 15±5°C. 3) CV8814 free base (28.8 kg, CHP Lot No. 2493-1903-00484), EDC (26.7 kg, CHP Lot No. 147-190213), DMAP (1.70 kg, CHP Lot No. 152-190214) and DCM (306.6 kg, CHP Lot No. 328-190326) were charged to reactor R-402. 4) The contents of R-402 were stirred for at least 20 minutes. 5) The contents of R-402 were transferred to R-401 over a period of at least 30 minutes, maintaining the temperature below 25°C. 6) T max =20.0℃ 7) The temperature of the contents of R-401 was adjusted to 20±5°C and stirred for at least 16 hours. 8) After approximately 16 hours, the contents of R-401 were sampled. 9) Analysis of the sample by QC showed that 0% (0.05%) of CV-8814 free base (CV8814 free base) was detected relative to CV8972 (specification: ≦1% CV8814 free base). 10) The contents of R-401 were adjusted to 10±5°C. 11) Water (29.2 kg, CHP Lot No. 329-190326) was added slowly while maintaining the temperature below 25°C. 12)T max =12.3℃ 13) The temperature of the contents of R-401 was adjusted to 20±5°C, stirred for at least 15 minutes, and allowed to stand for at least 15 minutes. 14) The phases were separated. 15) The lower organic layer containing the product was transferred to reactor R-402. The aqueous layer was sent to a drum. 16) Water (29.0 kg, CHP Lot No. 329-190326) was charged to the reactor. 17) The biphasic mixture was stirred for 15 minutes and allowed to settle for 15 minutes. 18) The phases were separated. 19) The organic layer containing the product was transferred to reactor R-401. The aqueous layer was sent to a drum. 20) 8% aqueous NaHCO3 solution (sodium bicarbonate, 4.0 kg, CHP lot number 192-190221; water, 53.2 kg, CHP lot number 329-190326) was added to the reactor. 21) The mixture was stirred for at least 15 minutes and allowed to stand for at least 15 minutes. 22) The phases were separated. 23) The lower organic layer containing the product was transferred to reactor R-402. The aqueous layer was sent to a drum. 24) Water (29.0 kg, CHP Lot No. 329-190326) was added to the reactor. 25) The mixture was stirred for at least 15 minutes and allowed to stand for at least 15 minutes. 26) R-401 was cleaned with water (17.6 kg, CHP Lot No. 329-190326) and MEK (5.9 kg, CHP Lot No. 330-190326) and dried under a stream of N2. 27) The phases were separated. 28) The lower organic layer containing the product was transferred to reactor R-401. The aqueous layer was sent to a drum. 29) The contents of R-401 were concentrated under reduced pressure to approximately 72 L, maintaining the temperature below 45°C. 30)T max =32.0℃ 31) MEK (139.0 kg, CHP lot number 330-190326) was added to R-401. 32) The contents of R-401 were concentrated under reduced pressure to approximately 72 L, maintaining the temperature below 45°C. 33)T max =32.0℃ 34) MEK (139.1 kg, CHP lot number 330-190326) was added to R-401. 35) The contents of R-401 were concentrated under reduced pressure to approximately 72 L, maintaining the temperature below 45°C. 36)T max =29.2℃. 37) To determine the DCM:MEK ratio, 1H NMR of FiO was measured. DCM:MEK = 1:214.9 38) MEK (185.5 kg, CHP lot number 330-190326) was added to R-401. 39) MEK (208.7 kg, CHP Lot No. 330-190326) and concentrated HCl (30.2 kg, CHP Lot No. 274-190311) were charged to the cleaned R-402. 40) The temperature of the reactor contents was adjusted to 25±5°C. 41) The contents of R-401 were transferred to R-402 over approximately 1 hour, maintaining the temperature below 35°C. 42)T max =27.2℃ 43) The temperature of the contents of R-402 was heated to 50±5°C and stirred for at least 1 hour. 44) The temperature of the reactor contents was cooled to 20±5°C over 2 hours. 45) The contents of the reactor were stirred at 20±5° C. for 15 hours. 46) The solid was filtered. 47) The filter cake was rinsed with MEK (58.0 kg, CHP Lot No. 330-190326). 48) The filter cake was rinsed with MEK (58.0 kg, CHP Lot No. 330-190326). 49) The wet cake was dried in a tray oven at 20-25°C without a nitrogen bleed for at least 16 hours. 50) For KF, an IPC sample (IPC sample: 2479-1903-00489-87-01) was submitted to QC. Moisture content (standard: cKF ≦4%): 3.3% 51) The product CV8972 monohydrate was double wrapped, placed in an S-shape and weighed. 52) Analysis of dry matter (IPC sample: 2479-1903-00489-87-01): Appearance: White to off-white solid Weight: 48.6 kg (96.4% yield) · 1H NMR: Consistent with structure. · HPLC purity (area %): 99.1% GC analysis of residual solvents in FIO: 2-MeTHF: no peak DCM: no peak MTBE: no peak Heptane: no peak MEK: 343 ppm Acetic acid: 874 ppm

[0237] Step 3, the formation of CV-8972 (2479-1904-00494), was carried out according to Scheme 4. Scheme 4: [ka] [ka]

[0238] Production details for step 3 are provided in Table 19. [Table 19]

[0239] 1) CV8972 monohydrate (48.5 kg, CHP lot number 2479-1903-00489), water (48.5 kg, CHP lot number 329-190326), methanol (19.2 kg, CHP lot number 379-190404), and MEK (39.0 kg, CHP lot number 330-190326) were each charged into reactor R-402. 2) The contents of R-402 were adjusted to 20°C ± 5°C and stirred until a solution was obtained. 3) The contents of R-402 were passed through a 0.45 micron in-line filter and transferred to R-401. 4) Water (24.0 kg, CHP Lot No. 329-190326) and methanol (19.2 kg, CHP Lot No. 379-190404) were charged to R-402, passed through a 0.45 micron in-line filter, and transferred to R-401. 5) Methanol (192.1 kg, CHP Lot No. 379-190404) was passed through a 0.45 micron in-line filter and charged to the R-401. 6) The temperature of the contents of R-401 was adjusted to 60±5°C. 7) While maintaining the temperature at 60±5° C., MEK (899.6 kg, CHP Lot Nos. 330-190326; 380-190404) was charged to the R-401 over approximately 2 hours through a 0.45 micron in-line filter. 8) The contents of R-401 were stirred at 60±5° C. for at least 4 hours. 9) The temperature of the R-401 was adjusted to 20±5°C over a period of at least 3 hours. 10) The contents of R-401 were stirred at 20±5° C. for approximately 9 hours. 11) The contents of R-401 were pumped into the filter. 12) The filter cake was rinsed with MEK (105.4 kg, CHP Lot No. 380-190404). 13) The filter cake was rinsed with MEK (105.4 kg, CHP Lot No. 380-190404). 14) The wet cake was dried on the filter under vacuum for at least 30 minutes. 15) The wet cake was sealed in a filter dryer and dried under vacuum at ≦30° C. for at least 12 hours. 16) An IPC sample (IPC sample: 2479-1904-00494-32-01) was submitted to QC for KF and GC of residual solvents. Moisture content (standard: 2.8-3.8% according to cKF) = 3.5% GC analysis of residual solvents (specifications: MeOH≦3000 ppm; MEK≦5000 ppm) MeOH=215 ppm; MEK=185 ppm 17) The product CV-8972 was double wrapped, formed into an S shape, and weighed. 18) Analysis of dry matter (IPC sample: 2479-1904-00494-32-01): Appearance: White to off-white solid Weight = 41.7 kg (86.0% yield) · HPLC purity (area %)=99.9% Known Impurities: Nicotinic acid: no peak DMAP: No peak CV8814:0.1% 2,3,4-trimethoxybenzaldehyde: no peak Trimetazidine: No peak CV-10099: No peak CV-10046: No peak XRPD: Consistent with Form A Chloride ion content: 19.4% 1H NMR: Consistent with structure

[0240] conclusion The results presented above demonstrate that CV-8972 can be synthesized using Scheme 1. Reductive amination in Step 1 using starting materials 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol, sodium triacetoxyborohydride (STAB) as the reducing agent, catalytic acetic acid (AcOH) and 2-methyltetrahydrofuran (2-MeTHF) afforded CV-8814 free base (CV8814 free base) in 78.0% yield with 100.0% purity by HPLC after aqueous workup, solvent exchange, and crystallization. A 5 kg portion of CV-8814 free base (CV8814 free base) was diverted from the synthesis for publication. Step 2 coupling of CV8814 free base with nicotinic acid in DCM in the presence of EDC and catalytic DMAP proceeded to complete conversion to CV8972 free base by HPLC IPC. Solvent exchange into MEK and addition of concentrated HCl in MEK afforded CV8972 monohydrate in 96.4% yield with 99.1% purity by HPLC. The final form conversion in step 3 was accomplished by heating CV8972 monohydrate in a mixture of water, methanol, and MEK to 60°C ± 5°C and adding MEK to precipitate. White solid CV-8972 was obtained as Form A, confirmed by XRPD analysis, in 86.0% yield with 99.9% purity by HPLC. The overall yield of the GMP synthesis of CV-8972 was 64.7%. The final amount of CV-8972 produced was 41.7 kg.

[0241] Incorporation by Reference Throughout this disclosure, references and citations are made to other documents, such as patents, patent applications, patent publications, journals, books, articles, web content, etc. All such documents are incorporated herein by reference in their entirety for all purposes.

[0242] equivalent Various modifications of the invention and numerous further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the complete contents of this document, including the scientific and patent references cited herein. The subject matter herein contains important information, exemplification, and guidance that can be adapted to the practice of the invention in its various embodiments and equivalents thereof.

Claims

1. Compound of formula (X): 【Chemistry 26】 1. A method for preparing 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol are reacted to produce the free base form of the compound of formula (IX): 【Chemistry 27】 generating reacting the free base form of the compound of formula (IX) with nicotinic acid to produce the compound of formula (X). Including, The method does not include producing a salt form of the compound of formula (IX).

2. 10. The method of claim 1, wherein the step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol comprises sodium triacetoxyborohydride, acetic acid, and / or 2-methyltetrahydrofuran.

3. 3. The method of claim 2, wherein the step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol is carried out at 15° C. to 25° C., and the step of reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol does not include dichloromethane.

4. 2. The method of claim 1, wherein the step of reacting the free base form of the compound of formula (IX) with nicotinic acid produces the free base form of the compound of formula (X), and wherein the step of reacting the free base form of the compound of formula (IX) with nicotinic acid comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, 4-(dimethylamino)pyridine, and / or dichloromethane.

5. 5. The method of claim 4, wherein the step of reacting the free base form of the compound of formula (IX) with nicotinic acid is carried out at 20° C. to 25° C., and the method further comprises the step of converting the free base form of the compound of formula (X) to a salt form of the compound of formula (X).

6. 6. The method of claim 5, wherein the salt form of the compound of formula (X) is an HCl salt and / or a monohydrate.

7. 6. The method of claim 5, wherein the step of converting the free base form of the compound of formula (X) to the salt form of the compound of formula (X) comprises HCl and / or methyl ethyl ketone.

8. 6. The method of claim 5, wherein the step of converting the free base form of the compound of formula (X) to the salt form of the compound of formula (X) is performed at 50° C., the method further comprises converting the salt form of the compound of formula (X) from a first crystalline form to a second crystalline form, the step of converting the salt form of the compound of formula (X) from the first crystalline form to the second crystalline form comprises one selected from the group consisting of precipitating the salt form of the compound of formula (X), changing the solvent of the salt form of the compound of formula (X), and incubating the salt form of the compound of formula (X) at 60° C., and further wherein the method does not comprise the use of dioxane, ethyl acetate, or potassium carbonate.

9. Compound of formula (X): 【Chemistry 28】 1. A method for preparing Compound of formula (1): 【Chemistry 29】 with a compound of formula (2): 【Transformation 30】 to produce the free base form of the compound of formula (IX): 【Chemistry 31】 Steps to generate The free base form of the compound of formula (IX) can be converted into a compound of formula (3): 【Chemistry 32】 to produce the free base form of the compound of formula (X), and converting said free base form of said compound of formula (X) to the HCl salt of said compound of formula (X). Including, The method does not include producing a salt form of the compound of formula (IX).

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