Method for preparing metalloenzyme inhibitor compound

Abstract

Provided are a method for preparing a metalloenzyme inhibitor compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole, and an important intermediate for preparing the compound.

Description

Preparation method of metalloenzyme inhibitor compounds Technical Field

[0001] This disclosure relates to a method for preparing the metalloenzyme inhibitor compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzis[d]imidazole, and an important intermediate for preparing said compound. Background Technology

[0002] Compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole is a metalloenzyme inhibitor, specifically a selective inhibitor of aldosterone synthase CYP11B2, and shows promising application potential in the treatment of aldosterone-related diseases.

[0003] However, the existing methods for preparing this compound are complex and have low yields, which are not conducive to drug production. An improved preparation process is urgently needed. Summary of the Invention

[0004] This invention provides a method for preparing the metalloenzyme inhibitor compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole. More specifically, this invention provides a method for preparing the compound 6-(difluoromethyl)pyridazin-4-carboxylic acid, which is used to prepare 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole, and a novel intermediate structure.

[0005] This invention relates, in one aspect, to a method for preparing compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A), comprising the following steps:

[0006] (iv) N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazin-4-carboxamide (compound 11) was subjected to a cyclization condensation reaction under acidic conditions to form 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0007] In some specific embodiments, in step (iv), N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11) is dissolved in isopropyl acetate, and / or the acidic conditions are generated by adding an isopropanol solution of HCl, and / or the cyclization condensation reaction is carried out at a temperature of about 65 ± 5 °C.

[0008] In some specific embodiments, in step (iv), N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11) is dissolved in ethanol, and / or the acidic conditions are generated by adding an aqueous solution of HCl, and / or the cyclization condensation reaction is carried out at a temperature of about 60 ± 5 °C.

[0009] In some specific embodiments, the preparation method of compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A) further includes the following steps:

[0010] (iii) Make N 1 -Cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) or its hydrochloride salt reacts with 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) to form N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11).

[0011] In some specific embodiments, in step (iii), N is placed under an inert atmosphere. 1 The hydrochloride salt of cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) undergoes a coupling reaction with the reaction product of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) and CDI in isopropyl acetate to form N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11).

[0012] In some specific embodiments, in step (iii), N is introduced into the atmosphere at a temperature below room temperature and in an inert atmosphere. 1 Triethylamine and propylphosphonic anhydride were added to an ethyl acetate solution of cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) and 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7), and then heated to room temperature to react and form N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11).

[0013] In some specific embodiments, the preparation method of compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A) further includes the following steps:

[0014] (ii) Catalytic hydrogenation of N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9) to form N 1 -Cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) or its hydrochloride salt.

[0015] In some specific embodiments, in step (ii), a Pt-V / C catalyst is used to react with hydrogen in an isopropyl acetate solution of N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9) to form N 1 -Cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10);

[0016] The catalyst was removed by filtration, and then an ethyl acetate solution of HCl was added to precipitate a solid, thus obtaining N. 1 The hydrochloride salt of cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10).

[0017] In some specific embodiments, in step (ii), a Pt / C catalyst is used to react with hydrogen in an ethyl acetate solution of N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9) to form N 1 -Cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10).

[0018] In some specific embodiments, the preparation method of compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A) further includes the following steps:

[0019] (i) Reacting 1,2,4-trifluoro-5-nitrobenzene (compound 8) with cyclopropylamine to form N-cyclopropyl-4,5-difluoro-2-nitrobenzene (compound 9).

[0020] In some specific embodiments, in step (i), 1,2,4-trifluoro-5-nitrobenzene (compound 8), potassium carbonate, and cyclopropylamine are reacted in toluene at a temperature of about 105 ± 5 °C to form N-cyclopropyl-4,5-difluoro-2-nitrobenzylamine (compound 9).

[0021] In some specific embodiments, after N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9) is formed in toluene, water is added to extract and separate the organic phase, then glacial acetic acid is used to replace the solvent, and then water is added at room temperature to crystallize and separate the N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9).

[0022] In some specific embodiments, in step (i), 1,2,4-trifluoro-5-nitrobenzene (compound 8), potassium carbonate and cyclopropylamine are reacted in tetrahydrofuran at a temperature of about 0°C to room temperature to form N-cyclopropyl-4,5-difluoro-2-nitrobenzylamine (compound 9).

[0023] In some specific embodiments, the preparation method of compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzi[d]imidazole (compound A) further includes the synthesis of 6-(difluoromethyl)pyridazin-4-carboxylic acid (compound 7). Specifically, the preparation method of 6-(difluoromethyl)pyridazin-4-carboxylic acid (compound 7) includes the following steps:

[0024] (d) 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid ethyl ester (compound 5) is reduced and hydrolyzed to form 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

[0025] In some specific implementations, step (d) includes the following steps:

[0026] (d-1) In a mixed solution of THF and water, ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5) is hydrolyzed under alkaline conditions to form 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6); preferably, the alkaline conditions are generated by adding an aqueous solution of potassium carbonate;

[0027] (d-2) 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6) is reacted with formic acid and sodium formate in aqueous solution under the catalysis of Pd / C catalyst to form 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

[0028] In some specific implementations, step (d) includes the following steps:

[0029] (d-1') In a mixed solution of ethanol and triethylamine, ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5) was hydrogenated under the catalysis of a Pd / C catalyst to form ethyl 6-(difluoromethyl)pyridazine-4-carboxylate (compound 6-2).

[0030] (d-2') In a mixed solution of THF and water, ethyl 6-(difluoromethyl)pyridazine-4-carboxylate (compound 6-2) is hydrolyzed under alkaline conditions to form 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7); preferably, the alkaline conditions are generated by adding an aqueous solution of lithium hydroxide.

[0031] In some specific embodiments, the preparation method of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) further includes the following steps:

[0032] (c) 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4) is subjected to a substitution reaction to form 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5).

[0033] In some specific embodiments, in step (c), dimethylformamide and oxaloyl chloride are added to a dichloromethane solution of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4), and the reaction is carried out at a temperature of about 30±5°C to form ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5).

[0034] In some specific embodiments, in step (c), phosphorus oxychloride is added to a toluene solution of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4), and the reaction is carried out at a temperature of about 65±5°C to form ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5).

[0035] In some specific embodiments, the preparation method of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) further includes the synthesis of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4). Specifically, the preparation method of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4) includes the following steps:

[0036] (b) In an ethanol solvent, under acidic conditions, diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3) was reacted with hydrazine by heating under reflux to form ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4).

[0037] In some specific embodiments, in step (b), the hydrazine and the acidic conditions are provided by using hydrazine bisulfate.

[0038] In some specific embodiments, in step (b), the hydrazine and the acidic conditions are provided by using hydrazine monohydrochloride.

[0039] In some specific embodiments, the preparation methods of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) and ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4) further include the following steps:

[0040] (a) Ketopropyl diethyl malonate (compound 1) and 1,1-difluoroacetone (compound 2) were dissolved in tetrahydrofuran at a temperature below room temperature. Ketopropyl diethyl malonate (compound 1) was reacted with 1,1-difluoroacetone (compound 2) under alkaline conditions at room temperature to form 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3).

[0041] In some specific embodiments, in step (a), the alkaline conditions are provided by using triethylamine.

[0042] In some specific embodiments, in step (a), the basic conditions are provided by using diazabicyclo[2.2.2]octane.

[0043] It should be understood that the above-mentioned methods for preparing 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) and ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4) can be carried out separately for preparing compounds 7 and 4, or can be used as part of the method for preparing 1-cyclopropyl-2-(6-(difluoromethyl)pyridazine-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A) for preparing compound A.

[0044] Another aspect of the present invention relates to a compound having the following structure:

[0045] That is, N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11).

[0046] The present invention also relates to the use of N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazin-4-carboxamide (compound 11) in the preparation of 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0047] Another aspect of the present invention relates to a compound having the following structure:

[0048] That is, diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3).

[0049] The present invention also relates to the use of diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3) in the preparation of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7). The present invention further relates to the use of diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3) in the preparation of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4).

[0050] Another aspect of the present invention relates to compounds having the following structure:

[0051] The use of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4) in the preparation of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

[0052] Another aspect of the present invention relates to compounds having the following structure:

[0053] The use of ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5) in the preparation of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

[0054] Another aspect of the present invention relates to the compound 6-(difluoromethyl)pyridazine-4-carboxylic acid, which is obtained by the preparation method according to the present invention.

[0055] Another aspect of the present invention relates to a compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole, said compound being obtained by the preparation method according to the present invention. In some specific embodiments, the compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole is prepared using 6-(difluoromethyl)pyridazin-4-carboxylic acid obtained by the preparation method according to the present invention.

[0056] Another aspect of the present invention relates to a pharmaceutical composition comprising:

[0057] (i) the compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole obtained by the preparation method according to the present invention, and

[0058] (ii) Pharmaceutically acceptable carriers, diluents or excipients.

[0059] In some embodiments, the pharmaceutical composition also includes additional therapeutic agents.

[0060] In some specific embodiments, the pharmaceutical composition may include additional therapeutic agents selected from anticancer agents, antifungal agents, cardiovascular therapeutic agents, anti-inflammatory agents, chemotherapeutic agents, anti-angiogenic agents, cytotoxic agents, antiproliferative agents, metabolic disease therapeutic agents, ophthalmic disease therapeutic agents, central nervous system (CNS) disease therapeutic agents, urinary disease therapeutic agents, and gastrointestinal disease therapeutic agents.

[0061] Another aspect of the present invention relates to the use of compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzimidazole, obtained by the preparation method according to the present invention, in pharmaceutical preparation.

[0062] In some specific embodiments, the drug is used to regulate the activity of metalloenzymes.

[0063] In some specific embodiments, the drug is used to regulate the activity of aldosterone synthase CYP11B2.

[0064] In some specific embodiments, the drug is used to treat metalloenzyme-related conditions or diseases, wherein the conditions or diseases are cancer, cardiovascular disease, endocrine disorders, fibrosis, kidney disease, inflammatory diseases, infectious diseases, gynecological diseases, metabolic diseases, ophthalmic diseases, central nervous system (CNS) diseases, urinary diseases, or gastrointestinal diseases.

[0065] In some specific embodiments, the condition or disease is adrenal carcinoma, adrenal adenoma, leukemia, breast cancer, hypertension, refractory hypertension, pulmonary hypertension, heart failure, diastolic dysfunction, left ventricular diastolic dysfunction, diastolic heart failure, systolic dysfunction, systolic heart failure, post-myocardial infarction syndrome, coronary artery disease, myocardial necrosis, atrial fibrillation, atherosclerosis, restenosis, endothelial dysfunction, vascular injury, myocardial infarction, left ventricular hypertrophy, vascular wall hypertrophy, endothelial thickening, arterial fibrinoid necrosis, vascular disease, and diseases associated with primary or secondary aldosteronism and adrenal hyperplasia. Diseases including diabetes, metabolic syndrome, insulin resistance, neuropathy, insulinopathy, diabetic nephropathy, diseases characterized by increased collagen formation, fibrosis and matrix remodeling after hypertension, diseases characterized by fibrosis and matrix remodeling after endothelial cell dysfunction, myocardial fibrosis, vascular fibrosis, renal failure, chronic renal failure, nephropathy, renal dysfunction, kidney disease, glomerulosclerosis, glomerulonephritis, nephrotic syndrome, polycystic kidney disease, hypokalemia, retinopathy, sleep apnea, obstructive sleep apnea, muscular dystrophy, stroke, liver disease, non-alcoholic steatohepatitis, cirrhosis or non-alcoholic fatty liver disease.

[0066] In a preferred embodiment, the condition or disease is hypertension, refractory hypertension, pulmonary hypertension, atherosclerosis, or hypokalemia.

[0067] Another aspect of the present invention relates to a treatment for a symptom or disease, comprising administering to a subject in need an effective amount of the compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzimidazole, prepared according to the method of the present invention. The symptom or disease is as described above. Attached Figure Description

[0068] Figure 1 shows the synthetic route for 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

[0069] Figure 2 shows another synthetic route for 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

[0070] Figure 3 shows the synthetic route for 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0071] Figure 4 shows another synthetic route for 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A). Detailed Implementation

[0072] definition

[0073] To facilitate a better understanding of this invention, certain terms are defined herein.

[0074] As used herein, the term "treatment" of a disease includes the prevention, improvement, reduction, and / or management of the disease and / or symptoms that may lead to the disease. The terms "treatment" and "management" refer to methods of reducing or alleviating a disease and / or its accompanying symptoms. According to this disclosure, "treatment" includes preventing, blocking, inhibiting, weakening, protecting, modulating, reversing, and reducing the effects of a disease, such as its harmful effects.

[0075] As used herein, “inhibition” includes prevention, reduction, and cessation of progression. The term “regulation” refers to an increase or decrease in enzyme activity in response to exposure to compounds of this disclosure.

[0076] The term "inhibitor" as used herein refers to a molecule that exhibits inhibition of metalloenzyme activity. "Inhibition" as used herein means a reduction in metalloenzyme activity compared to the activity of the metalloenzyme in the absence of an inhibitor. In some embodiments, the term "inhibition" means a reduction in metalloenzyme activity of at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95%. In other embodiments, inhibition means a reduction in metalloenzyme activity of about 5% to about 25%, about 25% to about 50%, about 50% to about 75%, or about 75% to 100%. In some embodiments, inhibition means a reduction in metalloenzyme activity of about 95% to 100%, for example, a reduction of 95%, 96%, 97%, 98%, 99%, or 100%. This reduction can be measured using various techniques recognized by those skilled in the art.

[0077] The terms “administration” or “application” refer to the means by which a compound is introduced into a target to achieve its intended function. Examples of possible routes of administration include injection (subcutaneous, intravenous, parenteral, intraperitoneal, intrathecal), topical, oral, inhalation, rectal, and transdermal administration.

[0078] The term "effective amount" refers to the amount of a compound that achieves the desired result within the necessary dosage and time period. The effective amount of a compound can vary depending on factors such as the subject's disease condition, age, and weight, as well as the compound's ability to elicit the desired response in the subject. Dosing regimens can be adjusted to provide the best therapeutic response. An effective amount is also the amount in which the beneficial therapeutic effect outweighs any toxic or harmful effects (e.g., side effects) of the inhibitory compound.

[0079] The phrases “systemic administration,” “systemic drug delivery,” “peripheral administration,” and “peripheral drug delivery” used here refer to the administration of a compound, drug, or other substance to the patient’s system, where it undergoes metabolism and other similar processes.

[0080] The term "therapeutic effective dose" refers to an amount of compound applied that is sufficient to prevent or, to some extent, alleviate the development of one or more symptoms of a disease or condition being treated.

[0081] The therapeutically effective amount (i.e., effective dose) of the compound can be from about 0.005 μg / kg body weight to about 200 mg / kg body weight, preferably from about 0.01 mg / kg body weight to about 200 mg / kg body weight, more preferably from about 0.015 mg / kg body weight to about 30 mg / kg body weight. In other embodiments, the therapeutically effective amount can be from about 1.0 pM to about 10 μM. Those skilled in the art will understand that certain factors can affect the dose required to effectively treat a subject, including but not limited to the severity of the disease or condition, prior treatment, the subject's overall health status and / or age, and other pre-existing conditions. Furthermore, treating a subject with a therapeutically effective amount of the compound can include monotherapy or preferably can include a series of treatments. In one example, a subject is treated with a compound at a concentration of about 0.005 μg / kg body weight to about 200 mg / kg body weight once daily for about 1 to 10 weeks, preferably 2 to 8 weeks, more preferably about 3 to 7 weeks, and even more preferably about 4, 5, or 6 weeks. In another example, in the case of a chronic condition or disease, the subject can be treated daily for several years. It should also be understood that the effective dose of a compound used for treatment can be increased or decreased during a particular treatment.

[0082] The term "object" refers to an animal, such as a mammal, including but not limited to primates (e.g., humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In some implementations, the object is humans.

[0083] As used herein, the term "pharmaceutically acceptable" means a carrier, loading agent, diluent, excipient, and / or salt that must be compatible with other components of the formulation and not be harmful to the recipient.

[0084] As used herein, the term "pharmaceuticalally acceptable carrier, diluent, or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that has been approved by the U.S. Food and Drug Administration for use in humans or animals and has no adverse effects on the composition of the pharmaceutical composition.

[0085] In this application (including the claims), terms used without a quantifier mean "one or more". Thus, for example, reference to "sample" includes multiple samples unless the context clearly indicates otherwise (e.g., multiple samples), etc.

[0086] Throughout the specification and claims, unless the context otherwise requires, the words “comprising,” “including,” and “containing” are used in a non-exclusive sense.

[0087] As used herein, the term “about” when referring to a value means including variations based on a specific amount, within ±20% in some embodiments, within ±10% in some embodiments, within ±5% in some embodiments, within ±1% in some embodiments, within ±0.5% in some embodiments, and within ±0.1% in some embodiments, because such variations are suitable for performing the disclosed methods or using the disclosed compositions.

[0088] It should be understood that the singular article “a” (corresponding to the English words “a,” “an,” and “the”) used in this disclosure and the appended claims includes plural objects unless otherwise expressly stated herein. Thus, a pharmaceutical composition comprising, for example, a “pharmaceutical-acceptable carrier, diluent, or excipient” includes one, or two or more, pharmaceutically acceptable carriers, diluents, or excipients.

[0089] The numerical limits or ranges stated in this article include endpoints, specifically all values ​​and subranges within the numerical limits or ranges.

[0090] The present invention will be further described below through specific embodiments. Unless otherwise specified, the terminology used herein has the same meaning as commonly understood by those skilled in the art.

[0091] This invention relates to compound (I) 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (hereinafter sometimes simply referred to as "compound A"). Compound A has the chemical formula C. 15 N4H 10 F4, molecular weight 322.3. Compound A is a metalloenzyme inhibitor, specifically a selective inhibitor of aldosterone synthase CYP11B2. The synthetic method and biological activity of compound A can be found, for example, in WO2018 / 125800, the entire contents of which are incorporated herein by reference.

[0092] In the preparation of compound A, compound 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) is an important intermediate.

[0093] For ease of description, some compounds involved in the preparation process of this invention are designated by numbers (Compound 1, Compound 2, and so on). The corresponding structures and chemical names of the numbered compounds are listed in Table 1 below:

[0094] Table 1 List of compounds

[0095] Preparation method of compound A

[0096] This invention provides a method for preparing compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A), comprising the following steps:

[0097] (iv) Compound 11 undergoes a cyclization condensation reaction under acidic conditions to form compound A.

[0098] In some preferred embodiments, in step (iv), compound 11 is dissolved in isopropyl acetate, and / or the acidic conditions are generated by adding an isopropanol solution of HCl, and / or the cyclization condensation reaction is carried out at a temperature of about 65 ± 5 °C.

[0099] In some specific embodiments, in step (iv), the compound 11 is dissolved in ethanol, and / or the acidic conditions are generated by adding an aqueous solution of HCl, and / or the cyclization condensation reaction is carried out at a temperature of about 60 ± 5 °C.

[0100] In some specific embodiments, the preparation method of compound A further includes the following steps:

[0101] (iii) React compound 10 or its hydrochloride salt with compound 7 to form compound 11.

[0102] In some preferred embodiments, in step (iii), the hydrochloride salt of compound 10 is coupled with the reaction product of compound 7 and CDI in isopropyl acetate under an inert atmosphere to form compound 11.

[0103] In some specific embodiments, in step (iii), triethylamine and propylphosphonic anhydride are added to an ethyl acetate solution of compound 10 and compound 7 at a temperature below room temperature and in an inert atmosphere, and then heated to room temperature to react and form compound 11.

[0104] In some specific embodiments, the preparation method of compound A further includes the following steps:

[0105] (ii) Catalytically hydrogenate compound 9 to form compound 10 or its hydrochloride salt.

[0106] In some preferred embodiments, in step (ii), a Pt-V / C catalyst is used to react with hydrogen in an isopropyl acetate solution of compound 9 to form compound 10. The catalyst is removed by filtration, and then an ethyl acetate solution of HCl is added to precipitate a solid to obtain the hydrochloride salt of compound 10.

[0107] In some specific embodiments, in step (ii), a Pt / C catalyst is used to react with hydrogen in an ethyl acetate solution of compound 9 to form compound 10.

[0108] In some specific embodiments, the preparation method of compound A further includes the following steps:

[0109] (i) React compound 8 with cyclopropylamine to form compound 9.

[0110] In some preferred embodiments, in step (i), compound 8, potassium carbonate, and cyclopropylamine are reacted in toluene at a temperature of about 105 ± 5 °C to form compound 9. In some specific embodiments, after compound 9 is formed in toluene, water is added to extract and separate the organic phase, followed by solvent replacement with glacial acetic acid, and then water is added at room temperature for crystallization to obtain compound 9.

[0111] In some specific embodiments, in step (i), compound 8, potassium carbonate and cyclopropylamine are reacted in tetrahydrofuran at a temperature of about 0°C to room temperature to form compound 9.

[0112] According to the preferred embodiment, in step (i), the desired isomer can be obtained as the main product in high yield, and can be separated by crystallization, facilitating subsequent steps; in step (i), by selecting a suitable catalyst type and reaction condition combination, the reduced unstable oily product can be obtained more selectively, and the product can be converted into a crystalline HCl salt that remains stable in an inert atmosphere, which is beneficial to improving the yield; steps (iii) and (iv) can be carried out continuously in a condensation process, further improving the yield of compound A while shortening the process cycle time; overall, the preferred preparation method of compound A can suppress or minimize the occurrence of side reactions in all steps, and does not require the use of chromatography for the purification and separation of intermediates, which is beneficial to large-scale industrial production. Therefore, the present invention develops an efficient, non-chromatographic, and scalable method for the preparation of compound A.

[0113] This invention also provides a method for purifying compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A), comprising the following steps:

[0114] (1) Dissolve solid compound A in an organic solvent selected from formic acid, acetic acid, propionic acid, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, diethyl sulfoxide, C1-C6 alcohols or polyols, or acetonitrile, or suspend it in such an organic solvent. If necessary, it may be heated appropriately to obtain a solution or suspension.

[0115] (2) Add water to the above solution or suspension to cause the solid to precipitate;

[0116] (3) The solid was then collected by filtration, washed with water and dried to obtain purified compound A.

[0117] In some specific embodiments, in step (1), the organic solvent is acetic acid, preferably anhydrous acetic acid; and / or, the heating temperature is 65±5℃.

[0118] In some specific embodiments, in step (2), depending on the miscibility of water with the organic solvent used, the amount of water is 0.1 to 100 times the volume of the organic solvent used, preferably 0.5 to 10 times.

[0119] In some specific embodiments, the purification method for compound A further includes cooling the mixture to room temperature and stirring it at room temperature for a period of time, such as about 2 hours, after the solid precipitates in step (2).

[0120] It should be understood that the purification method for compound A described above can be carried out alone or as part of the preparation method of the compound to prepare purified compound A.

[0121] In a preferred embodiment, the preparation method of compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A) includes the following steps:

[0122] (d-1) In a mixed solution of THF and water, ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5) was hydrolyzed under alkaline conditions to form 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6);

[0123] (d-2) 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6) was reacted with formic acid and sodium formate in aqueous solution under the catalysis of Pd / C catalyst to form 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7);

[0124] (iii) Make N 1 -Cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) or its hydrochloride salt reacts with 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) to form N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11);

[0125] (iv) N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazin-4-carboxamide (compound 11) was subjected to a cyclization condensation reaction under acidic conditions to form 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0126] In some specific embodiments, the reaction conditions and preparation methods of various reactants involved in the above steps are as described in other parts of this document.

[0127] Preparation method of intermediate compound 7

[0128] Another aspect of the present invention provides a method for preparing the intermediate 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) required for the preparation of compound A, comprising the following steps:

[0129] (d) Compound 5 is reduced and hydrolyzed to form compound 7.

[0130] In some preferred embodiments, step (d) includes the following steps:

[0131] (d-1) In a mixed solution of THF and water, compound 5 is hydrolyzed under alkaline conditions to form compound 6; preferably, the alkaline conditions are generated by adding an aqueous solution of potassium carbonate.

[0132] (d-2) Compound 6 is reacted with formic acid and sodium formate in aqueous solution under the catalysis of a Pd / C catalyst to form compound 7.

[0133] In some other implementations, step (d) includes the following steps:

[0134] (d-1') In a mixed solution of ethanol and triethylamine, compound 5 was hydrogenated under the catalysis of a Pd / C catalyst to form compound 6-2;

[0135] (d-2') In a mixed solution of THF and water, compound 6-2 is hydrolyzed under alkaline conditions to form compound 7; preferably, the alkaline conditions are generated by adding an aqueous solution of lithium hydroxide.

[0136] In some specific embodiments, the preparation method of compound 7 further includes the following steps:

[0137] (c) Compound 4 is subjected to a substitution reaction to form compound 5.

[0138] In some preferred embodiments, in step (c), dimethylformamide and oxaloyl chloride are added to a dichloromethane solution of compound 4 and reacted at a temperature of about 30 ± 5 °C to form compound 5.

[0139] In some specific embodiments, in step (c), phosphorus oxychloride is added to a toluene solution of compound 4 and reacted at a temperature of about 65 ± 5 °C to form compound 5.

[0140] In some specific embodiments, the preparation method of compound 7 further includes the following steps:

[0141] (b) Compound 3 was reacted with hydrazine in an ethanol solvent under acidic conditions by heating and reflux to form compound 4.

[0142] In some preferred embodiments, in step (b), the hydrazine and the acidic conditions are provided by using hydrazine bisulfate.

[0143] In some specific embodiments, in step (b), the hydrazine and the acidic conditions are provided by using hydrazine monohydrochloride.

[0144] In some specific embodiments, the preparation method of compound 7 further includes the following steps:

[0145] (a) Compound 1 and Compound 2 were dissolved in tetrahydrofuran at a temperature below room temperature, and Compound 1 and Compound 2 were reacted under alkaline conditions at room temperature to form Compound 3.

[0146] In some preferred embodiments, in step (a), the alkaline conditions are provided by using triethylamine.

[0147] In some specific embodiments, in step (a), the basic conditions are provided by using diazabicyclo[2.2.2]octane.

[0148] According to the preferred embodiment, steps (a)-(d) can significantly improve the yield of compound 7, shorten the process cycle time, and suppress or minimize the occurrence of side reactions. Therefore, the present invention develops an efficient and scalable method for preparing compound 7.

[0149] Preparation method of intermediate compound 4

[0150] Another aspect of the present invention provides a method for preparing the intermediate 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) required for the preparation of compounds A and 7, comprising the following steps:

[0151] (b) Compound 3 was reacted with hydrazine in an ethanol solvent under acidic conditions by heating and reflux to form compound 4.

[0152] In some specific embodiments, the preparation method of compound 4 further includes the following steps:

[0153] (a) Compound 1 and Compound 2 were dissolved in tetrahydrofuran at a temperature below room temperature, and Compound 1 and Compound 2 were reacted under alkaline conditions at room temperature to form Compound 3.

[0154] In some specific embodiments, the reaction conditions and preparation methods of various reactants involved in the above steps are as described in other parts of this document.

[0155] Compounds and their uses

[0156] The present invention also provides a series of novel intermediate compounds, and the use of the intermediate compounds in the preparation of compound A and / or compound 7.

[0157] On the one hand, the present invention also provides a novel compound N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11).

[0158] In another aspect, the present invention also provides the use of N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazin-4-carboxamide (compound 11) in the preparation of 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0159] In another aspect, the present invention also provides 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7), 1,2,4-trifluoro-5-nitrobenzene (compound 8), N-cyclopropyl-4,5-difluoro-2-nitrobenzide (compound 9), N... 1 -Cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) and N 1 Use of any of the compounds in cyclopropyl-4,5-difluorophenyl-1,2-diamine(x) hydrochloride (hydrochloride of compound 10) in the preparation of 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0160] In another aspect, the present invention also provides a novel compound, diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3).

[0161] In another aspect, the present invention also provides the use of diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3) in the preparation of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4).

[0162] In another aspect, the present invention also provides the use of diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3) in the preparation of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

[0163] In another aspect, the present invention also provides the use of diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3) in the preparation of 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0164] In another aspect, the present invention also provides the use of diethyl ketomalonate (compound 1) and 1,1-difluoroacetone (compound 2) in the preparation of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4).

[0165] In another aspect, the present invention also provides the use of any one of the following compounds in the preparation of 6-(difluoromethyl)pyridinium malonate (compound 1), 1,1-difluoroacetone (compound 2), ethyl 3-hydroxy-6-(difluoromethyl)pyridinium-4-carboxylate (compound 4), ethyl 3-chloro-6-(difluoromethyl)pyridinium-4-carboxylate (compound 5), 3-chloro-6-(difluoromethyl)pyridinium-4-carboxylic acid (compound 6), and ethyl 6-(difluoromethyl)pyridinium-4-carboxylate (compound 6-2) in the preparation of 6-(difluoromethyl)pyridinium-4-carboxylic acid (compound 7).

[0166] In another aspect, the present invention also provides the use of any one of the following compounds in the preparation of 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-carboxylic acid, compound 1), 1,1-difluoroacetone (compound 2), ethyl 3-hydroxy-6-(difluoromethyl)pyridazin-4-carboxylic acid (compound 4), ethyl 3-chloro-6-(difluoromethyl)pyridazin-4-carboxylic acid (compound 5), ethyl 3-chloro-6-(difluoromethyl)pyridazin-4-carboxylic acid (compound 6) and ethyl 6-(difluoromethyl)pyridazin-4-carboxylic acid (compound 6-2) in the preparation of 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

[0167] Pharmaceutical Composition

[0168] The present invention also provides a pharmaceutical composition comprising compound A (1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole) prepared according to the method of the present invention.

[0169] Specifically, another aspect of the present invention relates to a pharmaceutical composition comprising:

[0170] (i) the compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole prepared according to the method of the present invention, and

[0171] (ii) Pharmaceutically acceptable carriers, diluents or excipients.

[0172] In some embodiments, the pharmaceutical composition also includes additional therapeutic agents.

[0173] In some specific embodiments, the pharmaceutical composition may include additional therapeutic agents selected from anticancer agents, antifungal agents, cardiovascular therapeutic agents, anti-inflammatory agents, chemotherapeutic agents, anti-angiogenic agents, cytotoxic agents, antiproliferative agents, metabolic disease therapeutic agents, ophthalmic disease therapeutic agents, central nervous system (CNS) disease therapeutic agents, urinary disease therapeutic agents, and gastrointestinal disease therapeutic agents.

[0174] Pharmaceutical uses of compound A

[0175] Another aspect of the present invention relates to the use of compound A (1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzimidazole) prepared according to the method of the present invention in pharmaceutical preparation.

[0176] In some specific embodiments, the drug is used to regulate the activity of metalloenzymes.

[0177] In some specific embodiments, the drug is used to regulate the activity of aldosterone synthase CYP11B2.

[0178] In some specific embodiments, the drug is used to treat metalloenzyme-related conditions or diseases, wherein the conditions or diseases are cancer, cardiovascular disease, endocrine disorders, fibrosis, kidney disease, inflammatory diseases, infectious diseases, gynecological diseases, metabolic diseases, ophthalmic diseases, central nervous system (CNS) diseases, urinary diseases, or gastrointestinal diseases.

[0179] In some specific embodiments, the condition or disease is adrenal carcinoma, adrenal adenoma, leukemia, breast cancer, hypertension, refractory hypertension, pulmonary hypertension, heart failure, diastolic dysfunction, left ventricular diastolic dysfunction, diastolic heart failure, systolic dysfunction, systolic heart failure, post-myocardial infarction syndrome, coronary artery disease, myocardial necrosis, atrial fibrillation, atherosclerosis, restenosis, endothelial dysfunction, vascular injury, myocardial infarction, left ventricular hypertrophy, vascular wall hypertrophy, endothelial thickening, arterial fibrinoid necrosis, vascular disease, and diseases associated with primary or secondary aldosteronism and adrenal hyperplasia. Diseases including diabetes, metabolic syndrome, insulin resistance, neuropathy, insulinopathy, diabetic nephropathy, diseases characterized by increased collagen formation, fibrosis and matrix remodeling after hypertension, diseases characterized by fibrosis and matrix remodeling after endothelial cell dysfunction, myocardial fibrosis, vascular fibrosis, renal failure, chronic renal failure, nephropathy, renal dysfunction, kidney disease, glomerulosclerosis, glomerulonephritis, nephrotic syndrome, polycystic kidney disease, hypokalemia, retinopathy, sleep apnea, obstructive sleep apnea, muscular dystrophy, stroke, liver disease, non-alcoholic steatohepatitis, cirrhosis or non-alcoholic fatty liver disease.

[0180] In a preferred embodiment, the condition or disease is hypertension, refractory hypertension, pulmonary hypertension, atherosclerosis, or hypokalemia.

[0181] Compound A, prepared according to the method of the present invention, is a regulator and inhibitor of aldosterone synthase CYP11B2. It can effectively and specifically target CYP11B2, and therefore can be used to prevent or treat various symptoms or diseases related to CYP11B2.

[0182] Treatment

[0183] The present invention also provides a treatment for a symptom or disease, comprising administering to a subject in need an effective amount of compound A (1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzimidazole) prepared according to the method of the present invention. The symptom or disease is as described above.

[0184] Example

[0185] The present invention will be described in detail below through embodiments, which are not intended to limit the invention. Unless otherwise specified, the experimental methods in the following embodiments are conventional methods. All reagents and equipment used in the following embodiments are commercially available products and have no special requirements.

[0186] Preparation method of intermediate 7 in Example 1

[0187] Intermediate compound 7 was prepared via the reaction pathway shown in Figure 1.

[0188] Step 1: Synthesis of Compound 3

[0189] Tetrahydrofuran (2.0V) was loaded into the reactor at 20°C and then cooled to 0°C. 1,1-Difluoroacetone (117.5 kg, 1.0 equivalent) was added at 0°C, followed by dropwise addition of triethylamine (1.0 equivalent) at 0°C. Then, diethyl ketomalonate (220.0 kg, 1.0 equivalent) was added dropwise to the resulting mixture at 0°C over 5 hours. The reaction mixture was then heated to 25°C and stirred for 20 hours. The reaction system was cooled to 0°C, and the reaction was terminated by dropwise addition of a 5% (w / w) aqueous NaCl solution while maintaining the temperature at 10°C to separate the layers. The organic layer was concentrated under reduced pressure, and THF was exchanged for ethanol by solvent displacement to obtain a solution of intermediate compound 3. The yield of compound 3 was determined to be 71.2%. This solution was used directly in step 2 without further separation.

[0190] Step 2: Synthesis of Compound 4

[0191] Ethanol (EtOH, 9.0 V) was charged into the reactor at 20 °C, and then a solution of compound 3 (542.0 kg in EtOH, containing 240.6 kg of compound 3) was added at 20 °C. Hydrazine bisulfate (1.0 equivalent) was added to the resulting mixture at 20 °C. The mixture was then heated under reflux and stirred for 20 hours. The reaction mixture was cooled to 25 °C and centrifuged to obtain a wet product (compound 4). The wet filter cake was washed with water (1.0 V). The separated product was dried under reduced pressure at 50 °C for 12 hours to obtain the crude product compound 4. The dried filter cake was slurried in methyl tert-butyl ether (MTBE), centrifuged, and washed with MTBE. Further vacuum drying at 50 °C for 12 hours yielded 111.5 kg of product compound 4 (HPLC purity: 99.4% area; overall yield of steps 1-2 approximately 39.8%).

[0192] Step 3: Synthesis of Compound 5

[0193] Dichloromethane (DCM, 9.5V) was charged into the reactor at 20°C. The temperature was adjusted to 30°C, and compound 4 (110.0 kg, 1.0 equivalent) was added to the reactor at 30°C, followed by N,N-dimethylformamide (0.05 equivalent). Oxaloyl chloride (1.2 equivalent) was then added to the reactor over 10 hours, and the resulting mixture was stirred for another 6 hours. The reaction mixture was then added to cold water (5.0V) at 0°C. The phases were separated, and the organic phase was washed with water (1×5.0V). The organic phase was filtered, the filtrate was concentrated to 2-3V, and DCM was exchanged for THF by solvent displacement. The resulting solution of compound 5 was used directly in the subsequent hydrolysis step (step 4) without further separation.

[0194] Step 4: Synthesis of Compound 6

[0195] Water (4.0V) was added to a reactor containing a THF solution of compound 5 obtained in step 4 at 25°C, followed by dropwise addition of an aqueous solution of potassium carbonate (1.2 equivalents) at 25°C. The resulting mixture was stirred at room temperature for 16 hours. DCM (3.0V) was added to the reaction mixture to separate the phases. The aqueous phase containing the product was collected to give 971.5 kg of an aqueous solution of compound 6 (overall yield of steps 3-4: 95.2%, HPLC area purity: 99.6%).

[0196] Step 5: Synthesis of Compound 7

[0197] An aqueous solution of compound 6 obtained in step 4 (485.4 kg aqueous solution, compound 6 content 50.0 kg) was added to the reactor at 25 °C, followed by the addition of sodium formate (4.0 equivalents) at 25 °C. A Pd / C catalyst (50% wet, 500.0 g, 10 wt%) was added to the reactor at 25 °C, followed by the dropwise addition of formic acid (2.0 equivalents) over 5 hours at 25 °C, with stirring at this temperature for 40 hours. The mixture was filtered through a diatomaceous earth pad. The diatomaceous earth pad was washed with water (2 × 1.0 V). The combined filtrate was then added dropwise to an aqueous solution of HCl at 0 °C (5.0 equivalents). The mixture was further stirred for 5 hours, filtered, and the resulting wet filter cake was washed with water (6 × 1.0 V). The wet filter cake was dried under reduced pressure at 50°C to obtain 31.25 kg of pale yellow powder compound 7 (HPLC area purity: 99.6%; content determination: 99.4% by weight; overall yield of steps 3-5: 75%).

[0198] The total yield of steps 1-5 above is 30%.

[0199] Other preparation methods of intermediate 7 in Comparative Example 1

[0200] In this example, intermediate compound 7 is prepared via the reaction pathway shown in Figure 2.

[0201] Step 1: Synthesis of Compound 3

[0202] Tetrahydrofuran (30 L) was added to the reactor at room temperature (approximately 30 °C) under an inert atmosphere, followed by diethyl ketomalonate 1 (5 kg, 1.0 equivalent). The container of diethyl ketomalonate was rinsed, and the rinsing solution was added to the reactor. The resulting solution was cooled to 0 °C, and 1,1-difluoroacetone (1.0 equivalent) was added to the reactor. The mixture was maintained at 0 °C for 15 minutes, and then 1,4-diazabicyclo[2.2.2]octane (DABCO) was added. The reaction mixture was then stirred at room temperature for approximately 7 hours. The progress of the reaction was monitored by GC. THF was removed by distillation under reduced pressure, and ethyl acetate (5 V) and water (8 V) were added to the residue, and the resulting mixture was stirred at room temperature for approximately 25 minutes. The layers were separated, and the aqueous phase was further extracted with ethyl acetate (5 V). The combined organic phases were distilled to dryness under reduced pressure to give crude compound 3.

[0203] Step 2: Synthesis of Compound 4

[0204] Ethanol (10 V) was added to crude compound 3 obtained in step 1 at room temperature, followed by hydrazine monohydrochloride (1.6 equivalents) and p-toluenesulfonic acid (0.1 equivalents). Ethanol (10 V) was added again to the mixture, and the mixture was heated under reflux for 25 hours, with the reaction progress monitored by GC. The reaction mixture was concentrated to 2 V by vacuum distillation (keeping the temperature <45 °C). The residue was cooled to room temperature (approximately 30 °C) and diluted with ethyl acetate (6.5 V) and water (6.5 V). The mixture was stirred at room temperature (25 min) to separate the layers (approximately 30 min), and the bottom aqueous layer was removed. The aqueous layer was further extracted with ethyl acetate (6.5 V). The combined organic layers were concentrated to the residue by vacuum distillation (keeping the temperature of the reaction mixture below 45 °C). Toluene (2.5 V) was added to the residue, and the residue was further distilled to dryness to remove residual water. This azeotropic drying was repeated with toluene (2.5 V) to give crude compound 4.

[0205] Step 3: Synthesis of Compound 5

[0206] At 20°C, crude compound 4 obtained in step 2 was dissolved in toluene (5V) in a reactor, phosphorus oxychloride (1.6V) was added, and the resulting mixture was heated at 60°C for approximately 5 hours. The reaction mixture was cooled to 5°C, diluted with dichloromethane (10.8V), and then pre-cooled (5°C) water (10.8V) was added. The mixture was stirred for approximately 25 minutes and then allowed to stand. The layers were separated, and the aqueous layer was further extracted with dichloromethane (3.3V). The combined organic layers were distilled to dryness under reduced pressure (keeping the temperature <45°C) to give crude compound 5, which was purified by silica gel chromatography to give 1.8 kg of product with an area purity of 96% by HPLC analysis. The overall yield was calculated to be 26.5% based on the starting compound 1 (diethyl ketomalonate).

[0207] Step 4: Synthesis of Compound 6-2

[0208] Compound 5 (17 g, 1.0 equivalent) was loaded into a round-bottom flask at room temperature, followed by the addition of ethanol (10 V), triethylamine (1 V), and 10% Pd / C catalyst (50% wet, 1.7 g, 10 wt%) under a nitrogen atmosphere. The resulting mixture was then hydrogenated (using a hydrogen balloon) at 30 °C for 3–4 hours (the reaction was monitored by TLC). The reaction mixture was inertized and passed through a diatomaceous earth pad. The mixture was filtered, and the filtrate was concentrated into a solid residue by distillation under reduced pressure. The residue was dissolved in ethyl acetate (5V) and water (5V), and stirred for 20-30 minutes to separate the layers. The aqueous layer was further extracted with ethyl acetate (3V), and the combined organic layers were distilled to dryness under reduced pressure to give 12g of compound 6-2 (crude yield approximately 83%, area purity of 91% by HPLC analysis).

[0209] Step 5: Synthesis of Compound 7

[0210] Tetrahydrofuran (8V) was added to the reactor, followed by compound 6-2 (2.8 kg, 1.0 equivalent). The mixture was stirred at 20°C to obtain a solution. Water (2V) was added, followed by lithium hydroxide monohydrate (3.0 equivalent). The resulting mixture was stirred at 20°C for 1–2 hours, and the reaction was analyzed by TLC. Water (2.5V) was added, the mixture was stirred for 15 minutes, and then methyl tert-butyl ether (8V) was added. The two-phase mixture was stirred at 20°C for 15 minutes, allowed to stand (25 minutes), and the layers were separated. The aqueous layer was cooled to 5°C, and the pH was adjusted to 2–4 with concentrated HCl, precipitating a solid. The slurry was stirred at 0°C for 1 hour, filtered under reduced pressure to obtain a solid, and washed with water (2V). The wet filter cake was further dried under vacuum in an oven at 50°C for 1–2 hours to obtain 1.8 kg of compound 7 (area purity of 99.4% and yield of 75% by HPLC analysis).

[0211] The total yield of steps 1-5 above is 16%.

[0212] Example 2: Preparation method of compound A

[0213] In this example, compound A is prepared via the reaction pathway shown in Figure 3.

[0214] Step 1: Synthesis of Compound 9

[0215] Toluene (2.0 V) was added to the reactor at 25 °C, followed by compound 8 (99.5 kg, 1.0 equivalent) and potassium carbonate (2.0 equivalent). The resulting mixture was heated to 100 °C, and then a toluene (2.0 V) solution of cyclopropylamine (C-PrNH2, 1.0 equivalent) was added at 100 °C. After the reaction was complete, the reaction mixture was cooled to 25 °C. Softened water (6.0 V) was added to the mixture, and the mixture was stirred at 25 °C for at least 0.5 h. The phases were separated, and the organic phase was concentrated to 3-4 V. Glacial acetic acid (3.4 V) was added to the residue, followed by water (2.6 V) added dropwise at 20 °C. The resulting slurry was stirred at 20 °C for 2 h. The product was harvested by centrifugation, and the wet filter cake was washed with water (4 V) to obtain 106 kg of wet solids of compound 9 (83% separation yield), with an area purity of 99.6% as determined by HPLC.

[0216] Step 2: Synthesis of Compound 10 Hydrochloride

[0217] Isopropyl acetate (IPAc, 5.0V) was added to the reactor, followed by the addition of wet compound 9 (106 kg, 1.0 equivalent) obtained in step 1 under an inert nitrogen atmosphere. Pt-V / C catalyst (5 wt%) was added to the reactor, and the mixture was stirred for 10 minutes. A hydrogen pressure of 1–5 atm was then applied, and the reaction mixture was stirred at 20°C for 10 hours. The reaction progress was monitored by HPLC. After reduction, the reaction mixture was filtered, and the catalyst solid residue was washed twice with IPAc (2 × 2V). The filtrates were combined and separated into layers. The organic phase was collected and cooled to 10°C. An ethyl acetate solution of HCl (4 mol / L, 2.5 equivalent) was then added dropwise to the organic extract at 10°C. The resulting slurry was stirred and maintained at 10°C for at least 4 hours. The product was collected by centrifugation, washed with IPAc (2V), and vacuum dried at 25°C under a nitrogen atmosphere for 8 hours to give 112.6 kg of compound 10 hydrochloride, with an area purity of 95% and a yield of approximately 91% as determined by HPLC.

[0218] Steps 3 and 4 (Stacking Process): Synthesis of Compound A

[0219] Isopropyl acetate (3V) was added to reactor A under a nitrogen atmosphere at 25°C, followed by compound 7 (60.0 g, 1.0 equivalent). Carbonyl diimidazole (1.03 equivalent) was then added to the resulting mixture under a N2 atmosphere at 25°C. The reactor temperature was adjusted to 35°C, and the mixture was stirred for 5 hours under a N2 atmosphere.

[0220] Under a nitrogen atmosphere at 10°C, IPAc (4.0 V) was added to another reactor B, followed by the addition of compound 10 hydrochloride (1.03 equivalents) under a N2 atmosphere at 10°C. The imidazole derivative solution formed in reactor A was then slowly added to the hydrochloride solution in reactor B at 10°C, while the reaction mixture was maintained under a N2 atmosphere. The mixture was stirred for 1 hour at 10°C under a N2 atmosphere. The reaction progress was monitored by HPLC. Immediately after the coupling reaction was complete, a solution of isopropanol (i-PrOH) containing HCl (4 mol / L, 2.0 equivalents) was added to the reaction mixture under a N2 atmosphere at 10°C. The mixture was then heated to 65°C under a N2 atmosphere to initiate the cyclization and condensation reactions.

[0221] The mixture was stirred at 65°C for 18 hours, and HPLC analysis showed that the reaction was complete. Isopropanol was concentrated, and then water (4V) was added to the mixture, followed by extraction with ethyl acetate (4V). After standing and separation, the organic layer was washed with water (3×1V), and the ethyl acetate was concentrated to give product compound A (88.0 g, with an area purity of 98.6% and a yield of approximately 90% as determined by HPLC).

[0222] Step 5: Purification of Compound A

[0223] The crude compound A obtained in steps 3-4 (100.0 g) was suspended in glacial acetic acid (6 V) and then heated to approximately 65 °C. Water (6 V) was slowly added to the resulting solution over 5 hours at 65 ± 5 °C. The mixture was then cooled to approximately 25 °C and stirred at this temperature for 2 hours. The slurry was filtered and washed with water (2 × 2 V) to give purified compound A (94 g) as a grayish-white crystalline solid (recovery 94%; HPLC area purity 99.65%).

[0224] The overall yield of steps 1-5 above is 64%.

[0225] Comparative Example 2: Other methods for preparing compound A

[0226] In this example, compound A is prepared via the reaction pathway shown in Figure 4.

[0227] Step 1: Synthesis of Compound 9

[0228] Under an inert atmosphere, potassium carbonate (2.0 equivalents) was added to a stirred THF (15V) solution of compound 8 (250 g, 1.0 equivalent), followed by dropwise addition of cyclopropylamine (1.2 equivalents) at 0°C. The reaction mixture was gradually heated to room temperature and stirred for 16 hours. After monitoring the consumption of the starting material by TLC, the reaction mixture was diluted with cold water (12V) at approximately 0°C and extracted with EtOAc (3 × 8V). The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: 2% EtOAc in hexane, v / v) to give compound 9 (100 g, 33% yield, 99.45% purity) as an orange solid.

[0229] Step 2: Synthesis of Compound 10

[0230] At room temperature and under an inert atmosphere, catalyst Pd / C (50% wet, 37 g, 10 wt%) was added to a stirred solution of compound 9 (150 g, 0.7 equivalents) in ethyl acetate (15 V) in an autoclave. The reaction mixture was stirred at room temperature under a hydrogen atmosphere (60 psi) for 4 hours. After monitoring feed consumption by TLC, the catalyst was separated by filtration. The filtrate was concentrated under reduced pressure to obtain crude compound 10 as a brown semi-solid. This crude product was used directly in the next step without further purification.

[0231] Step 3: Synthesis of Compound 11

[0232] Compound 7 (1.0 equivalent) and triethylamine (2.0 equivalent) were added to a stirred solution of compound 10 (75 g, 0.41 mol) in ethyl acetate (20 V) at 0 °C under an inert atmosphere. Propylphosphonic anhydride (T3P, 50% EtOAc solution, 2.5 equivalent) was added dropwise to the resulting mixture over 30 minutes at 0 °C. The reaction mixture was gradually heated to room temperature and stirred for 2 hours. After monitoring the consumption of the starting material by TLC and LC-MS, the reaction mixture was adjusted to alkaline (pH ~8) using saturated NaHCO3 solution and extracted with EtOAc (3 × 1 L). The combined organic extracts were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: 2% MeOH in CH2Cl2, v / v) to obtain a solid, which was further purified by resuspending in a mixture of CH2Cl2 (30 mL) and hexane (1 L). The slurry was filtered and dried under vacuum to give compound 11 as a pale yellow solid (104 g, 0.3 mol, yield 75%).

[0233] Step 4: Synthesis of Compound A

[0234] At room temperature, under an inert atmosphere, 6N HCl (15V) was added dropwise to a stirred solution of compound 11 (78g, 0.23mol) in ethanol (10V). The reactor was then placed in an oil bath at 60°C, and the mixture was stirred for 15 minutes. After the starting material was completely consumed as monitored by TLC, the reaction mixture was cooled to 0°C and then poured into a cold saturated NaHCO3 solution (pH ~8) and extracted with EtOAc (2 × 3L). The combined organic extracts were dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain a crude product. The above steps were repeated three times using 78g of compound 11, and the crude products obtained (total 240g) were combined and purified by silica gel column chromatography (eluent: 2% MeOH in CH2Cl2, v / v) to give compound A (278g, 0.86mol, yield 94%, HPLC area purity 98.75%) as a grayish-white solid.

[0235] The total yield of steps 1-4 above is 23%.

[0236] The foregoing description of exemplary embodiments of the present invention provides a concrete example, but the invention is not limited thereto. Those skilled in the art should understand that the above embodiments are merely illustrative, and the specific embodiments and examples of the present invention should not be considered as limiting the scope of the invention. The embodiments contain important additional information, illustrations, and guidance that can be practiced in various embodiments and equivalents of this disclosure. Changes and modifications to the embodiments are possible within the scope of the spirit of the invention, and such changes and modifications should fall within the protection scope of the present invention.

Claims

1. A method for preparing compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A), characterized in that, Includes the following steps: (d-1) In a mixed solution of THF and water, ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5) was hydrolyzed under alkaline conditions to form 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6); (d-2) 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6) was reacted with formic acid and sodium formate in aqueous solution under the catalysis of Pd / C catalyst to form 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7); (iii) reacting N 1 - cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) or its hydrochloride salt with 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) to form N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11 ); (iv) N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazin-4-carboxamide (compound 11) was subjected to a cyclization condensation reaction under acidic conditions to form 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

2. The preparation method according to claim 1, characterized in that, In step (iv), N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11) is dissolved in isopropyl acetate, and / or the acidic conditions are generated by adding an isopropanol solution of HCl, and / or the cyclization condensation reaction is carried out at a temperature of about 65 ± 5 °C.

3. The preparation method according to claim 1, characterized in that, In step (iv), N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11) is dissolved in ethanol, and / or the acidic conditions are generated by adding an aqueous solution of HCl, and / or the cyclization condensation reaction is carried out at a temperature of about 60 ± 5 °C.

4. The preparation method according to claim 1, characterized in that, In step (iii), N is placed in an inert atmosphere. 1 The hydrochloride salt of cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) undergoes a coupling reaction with the reaction product of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) and CDI in isopropyl acetate to form N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11).

5. The preparation method according to claim 1, characterized in that, In step (iii), N 1 - cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10) is reacted with an ethyl acetate solution of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7) by adding triethylamine and propylphosphonic anhydride and then heating to room temperature to form N-(2-(cyclopropylamino)-4,5-difluorophenyl)-6-(difluoromethyl)pyridazine-4-carboxamide (compound 11).

6. The preparation method according to any one of claims 1 to 5, characterized in that, It also includes the following steps: (ii) catalytic hydrogenation of N-cyclopropyl-4,5-difluoro-2-nitroaniline (Compound 9) to form N 1 - cyclopropyl-4,5-difluorophenyl-1,2-diamine (Compound 10) or its hydrochloride salt.

7. The preparation method according to claim 6, characterized in that, In step (ii), the Pt-V / C catalyst is used to react N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9) in isopropyl acetate solution with hydrogen gas to form N 1 - cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10); The catalyst was removed by filtration, and then an ethyl acetate solution of HCl was added to precipitate a solid, thus obtaining N. 1 The hydrochloride salt of cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10).

8. The preparation method according to claim 6, characterized in that, In step (ii), using a Pt / C catalyst, N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9) is reacted with hydrogen in an ethyl acetate solution to form N. 1 -Cyclopropyl-4,5-difluorophenyl-1,2-diamine (compound 10).

9. The preparation method according to any one of claims 1 to 8, characterized in that, It also includes the following steps: (i) Reacting 1,2,4-trifluoro-5-nitrobenzene (compound 8) with cyclopropylamine to form N-cyclopropyl-4,5-difluoro-2-nitrobenzene (compound 9).

10. The preparation method according to claim 9, characterized in that, In step (i), 1,2,4-trifluoro-5-nitrobenzene (compound 8), potassium carbonate and cyclopropylamine are reacted in toluene at a temperature of about 105 ± 5 °C to form N-cyclopropyl-4,5-difluoro-2-nitrobenzene (compound 9).

11. The preparation method according to claim 10, characterized in that, After N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9) was formed in toluene, the organic phase was separated by water extraction, glacial acetic acid was added to the organic phase, and then water was added at room temperature for crystallization to obtain N-cyclopropyl-4,5-difluoro-2-nitroaniline (compound 9).

12. The preparation method according to claim 9, characterized in that, In step (i), 1,2,4-trifluoro-5-nitrobenzene (compound 8), potassium carbonate and cyclopropylamine are reacted in tetrahydrofuran at a temperature of about 0°C to room temperature to form N-cyclopropyl-4,5-difluoro-2-nitrobenzylamine (compound 9).

13. A method for preparing compound 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7), characterized in that, Includes the following steps: (d-1) In a mixed solution of THF and water, ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5) is hydrolyzed under alkaline conditions to form 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6); preferably, the alkaline conditions are generated by adding an aqueous solution of potassium carbonate; (d-2) 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 6) is reacted with formic acid and sodium formate in aqueous solution under the catalysis of Pd / C catalyst to form 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

14. The preparation method according to claim 13, characterized in that, It also includes the following steps: (c) 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4) is subjected to a substitution reaction to form 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5).

15. The preparation method according to claim 14, characterized in that, In step (c), dimethylformamide and oxaloyl chloride are added to a dichloromethane solution of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4), and the reaction is carried out at a temperature of about 30±5°C to form ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5).

16. The preparation method according to claim 14, characterized in that, In step (c), phosphorus oxychloride is added to a toluene solution of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4), and the reaction is carried out at a temperature of about 65±5°C to form ethyl 3-chloro-6-(difluoromethyl)pyridazine-4-carboxylate (compound 5).

17. A method for preparing ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4), characterized in that, Includes the following steps: (b) In an ethanol solvent, under acidic conditions, diethyl 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3) was reacted with hydrazine by heating under reflux to form ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 4).

18. The preparation method according to claim 17, characterized in that, In step (b), the hydrazine and the acidic conditions are provided by using hydrazine bisulfate.

19. The preparation method according to claim 17, characterized in that, In step (b), the hydrazine and the acidic conditions are provided by using hydrazine monohydrochloride.

20. The preparation method according to any one of claims 17 to 19, characterized in that, It also includes the following steps: (a) Ketopropyl diethyl malonate (compound 1) and 1,1-difluoroacetone (compound 2) were dissolved in tetrahydrofuran at a temperature below room temperature. Ketopropyl diethyl malonate (compound 1) was reacted with 1,1-difluoroacetone (compound 2) under alkaline conditions at room temperature to form 2-hydroxy-2-(2-oxo-3,3-difluoropropyl)malonate (compound 3).

21. The preparation method according to claim 20, characterized in that, In step (a), the alkaline conditions are provided by using triethylamine.

22. The preparation method according to claim 20, characterized in that, In step (a), the basic conditions are provided by using diazabicyclo[2.2.2]octane.

23. A compound having the following structure:

24. Use of the compound of claim 23 in the preparation of 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole (compound A).

25. A compound having the following structure:

26. Use of the compound of claim 25 in the preparation of 6-(difluoromethyl)pyridazine-4-carboxylic acid (compound 7).

27. Use of the compound of claim 25 in the preparation of ethyl 3-hydroxy-6-(difluoromethyl)pyridazine-4-carboxylate (compound 3).

28. A compound 6-(difluoromethyl)pyridazine-4-carboxylic acid, characterized in that, The compound is obtained by the preparation method according to any one of claims 13 to 16.

29. A compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole, characterized in that, The compound is obtained by the preparation method according to any one of claims 1 to 12.

30. A pharmaceutical composition comprising: (i) the compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole as claimed in claim 29, and (ii) Pharmaceutically acceptable carriers, diluents or excipients.

31. The pharmaceutical composition of claim 30, further comprising an additional therapeutic agent.

32. The pharmaceutical composition of claim 30, wherein the additional therapeutic agent is selected from anticancer agents, antifungal agents, cardiovascular therapeutic agents, anti-inflammatory agents, chemotherapeutic agents, anti-angiogenic agents, cytotoxic agents, antiproliferative agents, metabolic disease therapeutic agents, ophthalmic disease therapeutic agents, central nervous system (CNS) disease therapeutic agents, urinary disease therapeutic agents, and gastrointestinal disease therapeutic agents.

33. Use of the compound 1-cyclopropyl-2-(6-(difluoromethyl)pyridazin-4-yl)-5,6-difluoro-1H-benzo[d]imidazole of claim 29 in pharmaceutical preparation.

34. The use according to claim 33, wherein the drug is used to regulate metalloenzyme activity.

35. The use according to claim 33, wherein the drug is used to regulate the activity of aldosterone synthase CYP11B2.

36. The use according to claim 33, wherein the medicament is used to treat metalloenzyme-related conditions or diseases, wherein the condition or disease is cancer, cardiovascular disease, endocrine disorder, fibrosis, kidney disease, inflammatory disease, infectious disease, gynecological disease, metabolic disease, ophthalmic disease, central nervous system (CNS) disease, urinary disease, or gastrointestinal disease.

37. The use according to claim 36, wherein the condition or disease is adrenal carcinoma, adrenal adenoma, leukemia, breast cancer, hypertension, refractory hypertension, pulmonary hypertension, heart failure, diastolic dysfunction, left ventricular diastolic dysfunction, diastolic heart failure, systolic dysfunction, systolic heart failure, post-myocardial infarction syndrome, coronary artery disease, myocardial necrosis, atrial fibrillation, atherosclerosis, restenosis, endothelial dysfunction, vascular injury, myocardial infarction, left ventricular hypertrophy, vascular wall hypertrophy, endothelial thickening, arterial fibrinoid necrosis, vascular disease, and primary or secondary hyperaldosteronism and adrenal hyperplasia. Related conditions, diabetes, metabolic syndrome, insulin resistance, neuropathy, insulinopathy, diabetic nephropathy, diseases characterized by increased collagen formation, fibrosis and matrix remodeling after hypertension, diseases characterized by fibrosis and matrix remodeling after endothelial cell dysfunction, myocardial fibrosis, vascular fibrosis, renal failure, chronic renal failure, nephropathy, renal dysfunction, kidney disease, glomerulosclerosis, glomerulonephritis, nephrotic syndrome, polycystic kidney disease, hypokalemia, retinopathy, sleep apnea, obstructive sleep apnea, muscular dystrophy, stroke, liver disease, non-alcoholic steatohepatitis, cirrhosis or non-alcoholic fatty liver disease.

38. The use according to claim 36, wherein the condition or disease is hypertension, refractory hypertension, pulmonary hypertension, atherosclerosis, or hypokalemia.

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