Pharmaceutical use of compound a or salt thereof

Abstract

The present invention relates to the technical field of chemical drugs, provides a pharmaceutical use of compound A or a salt thereof, and specifically relates to a use of compound A or the salt thereof in preparation of a drug for preventing or treating heart failure (HF)-related diseases.

Description

Pharmaceutical uses of a compound A or a salt thereof Technical Field

[0001] This invention belongs to the field of chemical pharmaceutical technology and provides a pharmaceutical use of compound A or its salt, specifically relating to the use of compound A or its salt in the preparation of a drug for the prevention or treatment of heart failure (HF)-related diseases. Background Technology

[0002] Aldosterone is a steroid hormone with mineralocorticoid activity. It is primarily produced in response to angiotensin II, adrenocorticotropic hormone (ACTH), and the zona glomerulosa of the adrenal gland, which increases serum potassium levels. Its main physiological role in the kidneys is to maintain sodium and potassium balance by regulating cation exchange (Na+ reabsorption and K+ secretion) in the distal nephron. However, aldosterone has also been shown to be a pro-inflammatory and pro-fibrotic hormone in the blood vessels, heart, and kidneys. The effects of aldosterone on gene expression are regulated through binding to the mineralocorticoid receptor (MR) and typical nuclear hormone receptor pathways.

[0003] CYP11B2 is primarily expressed in the zona glomerulosa of the adrenal cortex and is known as an enzyme catalyzing a series of reactions from 11-deoxycorticosterone (i.e., aldosterone precursor) to aldosterone. CYP11B2 inhibitors have been reported to inhibit aldosterone production in studies using enzymes and cultured cells, and have shown inhibitory and therapeutic effects in various experimental animal models. Furthermore, CYP11B2 inhibitors have been confirmed to reduce plasma and urinary aldosterone levels and have antihypertensive effects in patients with hypertension and primary aldosteronism. Finding a means to inhibit the biosynthetic pathway of aldosterone is a highly feasible approach to establishing effective treatments for various aldosterone-related diseases.

[0004] Chronic heart failure (CHF) is the final stage in the development of many cardiovascular diseases such as myocardial infarction, cardiomyopathy, and hypertensive heart disease. Activation of the neuroendocrine system leading to myocardial remodeling is a key factor in the occurrence and development of CHF, and aldosterone is one of the important factors inducing cardiac remodeling within the renin-angiotensin-aldosterone system (RAAS). CHF is the final stage of most cardiovascular diseases and is one of the major health problems threatening health worldwide, with broad prospects for drug research and application.

[0005] Aldosterone synthase (CYP11B2) plays a crucial role in the pathogenesis of chronic heart failure (CHF), primarily by regulating aldosterone synthesis and thus exerting organic effects on the heart. The adverse effects of aldosterone on myocardial remodeling, particularly its promotion of fibrosis in the extracellular matrix of cardiomyocytes, are independent of and additive to the effects of Ang II.

[0006] PCT / CN2024 / 142727 discloses an aldosterone synthase inhibitor, its preparation method and uses, and discloses the use of the aldosterone synthase inhibitor in drugs for the prevention or treatment of CYP11B2-related diseases. Finding a small molecule compound of aldosterone synthase inhibitor with excellent efficacy in the treatment of chronic heart failure has become an urgent problem to be solved in this field. Summary of the Invention

[0007] In view of the problems existing in the prior art, the present invention provides a pharmaceutical use of compound A or its salt to solve the problems existing in the prior art.

[0008] This invention is achieved through the following technical solution:

[0009] This invention provides the use of compound A or a salt thereof in the preparation of a medicament for the prevention or treatment of heart failure (HF)-related diseases, characterized in that the structural formula of compound A is:

[0010] Furthermore, as a preferred embodiment of the present invention, the heart failure-related disease is selected from: chronic heart failure.

[0011] Furthermore, as a preferred embodiment of the present invention, the chronic heart failure is selected from congestive heart failure (CHF).

[0012] Furthermore, as a preferred embodiment of the present invention, the chronic heart failure is selected from heart failure with reduced ejection fraction (HFrEF).

[0013] Furthermore, as a preferred embodiment of the present invention, the chronic heart failure is selected from: heart failure with improved ejection fraction (HFimpEF).

[0014] Furthermore, as a preferred embodiment of the present invention, the chronic heart failure is selected from: heart failure with mildly reduced ejection fraction (HFmrEF).

[0015] Furthermore, as a preferred embodiment of the present invention, the chronic heart failure is selected from heart failure with preserved ejection fraction (HFpEF).

[0016] Further, as a preferred embodiment of the present invention, the salt is selected from hydrochloride, hydrobromide, phosphate, nitrate, sulfate, acetate, propionate, malonate, succinate, valerate, glutarate, adipate, oxalate, L-proline, lactobionate, glycine, alanine, arginine, lactate, cinnamate, fumarate, mandelate, maleate, hippurate, tartrate, citrate, malate, succinate, 2-naphthalenesulfonate, 1,5-naphthalenedisulfonate, camphorsulfonate, benzoate, salicylate, benzenesulfonate, methanesulfonate, or p-toluenesulfonate; preferably, each salt is independently selected from: hydrochloride, hydrobromide, phosphate, sulfate, benzenesulfonate, p-toluenesulfonate, maleate, fumarate, oxalate, succinate, or adipate.

[0017] Further, as a preferred embodiment of the present invention, the molar ratio of compound A to acid molecules in the salt is 1:0.3 to 1:3.5; preferably, the molar ratio of compound A to acid molecules is 1:1, 1:2, 1:3, 2:1, or 3:1; more preferably, the molar ratio of compound A to acid molecules is 1:1.

[0018] Further, as a preferred embodiment of the present invention, the amount of compound A or its salt (calculated as compound A) used in the drug is 0.25 mg to 50 mg; preferably, the amount of compound A or its salt (calculated as compound A) used in the drug is 0.25 mg to 25 mg; more preferably, the amount of compound A or its salt (calculated as compound A) used in the drug is 0.25 mg to 10 mg; most preferably, the amount of compound A or its salt (calculated as compound A) used in the drug is 0.25 mg, 0.5 mg, 0.75 mg, 1 mg, 2 mg, 3 mg, 4 mg, 6 mg, 7 mg, 8 mg, 9 mg, or 10 mg.

[0019] Furthermore, as a preferred embodiment of the present invention, the compound A or its salt is crystalline, amorphous, or a mixture thereof.

[0020] Furthermore, as a preferred embodiment of the present invention, one or more hydrogen atoms of the compound A or its salt are substituted with the isotope deuterium.

[0021] Furthermore, as a preferred embodiment of the present invention, the frequency of administration of the drug is selected from: once a day, twice a day, three times a day, once every two days, once every three days, and once a week. Preferably, the frequency of administration of the drug is selected from: once a day.

[0022] Furthermore, as a preferred embodiment of the present invention, the present invention also provides a method for preventing and / or treating heart failure-related diseases, comprising the following steps: administering to a patient in need a therapeutically effective amount of the compound A described herein or a pharmaceutically acceptable salt thereof. Preferably, the heart failure-related diseases are selected from: chronic heart failure, congestive heart failure (CHF), heart failure with reduced ejection fraction (HFrEF), heart failure with improved ejection fraction (HFimpEF), heart failure with mildly reduced ejection fraction (HFmrEF), and heart failure with preserved ejection fraction (HFpEF). For clarity, the general terms used in the description of the compounds are defined herein.

[0023] Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient. The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0024] The term "medicinal salt" refers to a salt of the compound of the present invention, prepared by reacting a compound having specific substituents discovered in the present invention with a medicinal acid or base.

[0025] In addition to the salt form, the compounds provided by this invention also exist in prodrug form. The prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to be converted into the compounds of this invention. Furthermore, the prodrugs can be converted into the compounds of this invention in the in vivo environment via chemical or biochemical methods.

[0026] Some compounds of this invention may exist in non-solventized or solvated forms, including hydrated forms. Generally, solvated and non-solventized forms are equivalent and both are included within the scope of this invention.

[0027] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0028] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the desired enantiomer in pure form. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0029] The atoms in the compounds of this invention are isotopes. Isotope derivatization can typically prolong half-life, reduce clearance rate, stabilize metabolism, and enhance in vivo activity. Furthermore, one embodiment is included, wherein at least one atom is replaced by an atom having the same number of atoms (protons) but different mass numbers (protons and neutrons). Examples of isotopes included in the compounds of this invention include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, and chlorine atoms, each comprising... 2 H, 3 H, 13 C 14 C 15 N、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 36 Cl. In particular, radioactive isotopes that emit radiation as they decay, such as 3 H or 14 C can be used for local anatomical examination of pharmaceutical preparations or compounds in vivo. Stable isotopes neither decay nor change with quantity and are not radioactive, therefore they can be used safely. When the atoms constituting the compounds of this invention are isotopes, the isotopes can be converted according to common methods by replacing the reagents used in the synthesis with reagents containing the corresponding isotopes.

[0030] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium. 2 H), Iodine-125 125 I) or C-14 14C). All isotopic variations of the compounds of the present invention, regardless of radioactivity, are included within the scope of the present invention.

[0031] Furthermore, one or more hydrogen atoms in the compound of the present invention are replaced by the isotope deuterium (2H). After deuteration, the compound of the present invention has the effects of prolonging the half-life, reducing the clearance rate, stabilizing metabolism, and improving in vivo activity.

[0032] The preparation methods of the isotope derivatives typically include phase-transfer catalysis. For example, a preferred deuteration method employs a phase-transfer catalyst (e.g., tetraalkylammonium salt, NBu4HSO4). Using a phase-transfer catalyst to exchange the methylene protons of a diphenylmethane compound results in the introduction of higher levels of deuterium than reduction with deuterated silanes (e.g., triethyldeuterated silane) in the presence of an acid (e.g., methanesulfonic acid) or with Lewis acids such as aluminum trichloride using sodium deuterated borate.

[0033] The term "pharmaceutically acceptable carrier" refers to any formulation carrier or medium capable of delivering an effective amount of the active substance of this invention without interfering with the biological activity of the active substance and without toxic side effects on the host or patient. Representative carriers include water, oil, vegetables and minerals, ointment bases, lotion bases, and ointment bases. These bases include suspending agents, thickeners, and transdermal penetration enhancers. Their formulations are well known to those skilled in the art of cosmetics or topical pharmaceuticals. For further information on carriers, see Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0034] The term "excipient" generally refers to the carrier, diluent, and / or medium required to formulate an effective pharmaceutical composition.

[0035] For pharmaceuticals or pharmacologically active agents, the term "effective amount" or "therapeutic effective amount" refers to a sufficient quantity of a drug or agent that is non-toxic but achieves the desired effect. For the oral dosage forms of this invention, the "effective amount" of one active substance in the composition refers to the quantity required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. A suitable effective amount in any given case can be determined by a person skilled in the art through routine testing.

[0036] The terms “active ingredient,” “therapeutic agent,” “active substance,” or “active agent” refer to a chemical entity that can effectively treat a target disorder, disease, or symptom.

[0037] "Optional" or "optionally" means that the event or condition described below may occur but is not required to occur, and the description includes both the scenario in which said event or condition occurs and the scenario in which said event or condition does not occur. Attached Figure Description

[0038] 1) Figure 1 shows the results of left ventricular ejection fraction by echocardiography of the compound of the present invention in an animal model of chronic heart failure. Detailed Implementation

[0039] The present invention will be further described in detail below with reference to the embodiments, but the content of the invention is not limited to the embodiments.

[0040] Example 1

[0041] Synthesis of (R)-N-(4-(2-cyanoquinoline-6-yl)-5,6,7,8-tetrahydroisoquinoline-8-yl)propionamide

[0042] Detailed operation steps:

[0043] Step A: Synthesis of ethyl 5-bromo-4-methylnicotinate

[0044] 5-Bromo-4-methylnicotinic acid (50.0 g, 231.45 mmol) and iodoethane (39.7 g, 254.59 mmol) were dissolved in 500 mL of N,N-dimethylformamide, and potassium bicarbonate (46.3 g, 462.90 mmol) was added. The mixture was degassed and protected with nitrogen, and the reaction was stirred at room temperature for 12 hours.

[0045] After the reaction was complete, the mixture was filtered, water was added to the filtrate, and the mixture was extracted with ethyl acetate (300 mL × 3 times). The combined organic phases were washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane = 1 / 10) to give 54.6 g of ethyl 5-bromo-4-methylnicotinate. [M+H] + =244.05.

[0046] Step B: Synthesis of methyl 4-bromo-8-oxo-5,6,7,8-tetrahydroisoquinoline-7-carboxylate

[0047] At -78°C, LDA (123 mL, 246.06 mmol, 2M) was added dropwise to a tetrahydrofuran (500 mL) solution of ethyl 5-bromo-4-methylnicotinate (54.6 g, 223.69 mmol). The mixture was stirred for 30 minutes, followed by the addition of a tetrahydrofuran (200 mL) solution of methyl acrylate (48.1 g, 559.22 mmol). The mixture was then stirred at -78°C for 2 hours.

[0048] After the reaction was complete, 400 mL of 10% acetic acid aqueous solution was added to the mixture to quench the reaction. The organic solvent was removed by evaporation, and the mixture was extracted with ethyl acetate (300 mL × 3 times). The combined organic phases were washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane = 1 / 10) to give 31.5 g of methyl 4-bromo-8-oxo-5,6,7,8-tetrahydroisoquinoline-7-carboxylate. [M+H] + =284.06.

[0049] Step C: Synthesis of 4-bromo-6,7-dihydroisoquinoline-8(5H)-one

[0050] 31.5 g (110.87 mmol) of methyl 4-bromo-8-oxo-5,6,7,8-tetrahydroisoquinoline-7-carboxylate was dissolved in 300 mL of hydrochloric acid (6 M), and the mixture was refluxed at 105 °C for 16 hours.

[0051] After the reaction was complete, the solvent was evaporated, 300 mL of water was added, and the pH was adjusted to approximately 9 with 1 N sodium hydroxide aqueous solution. The mixture was extracted with ethyl acetate (200 mL × 3 times), the organic phases were combined, washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane = 1 / 8) to give 19.6 g of 4-bromo-6,7-dihydroisoquinoline-8(5H)-one. [M+H] + =226.05.

[0052] Step D: Synthesis of (S)-N-(4-bromo-6,7-dihydroisoquinoline-8(5H)-ylidene)-2-methylpropane-2-sulfonamide

[0053] 4-Bromo-6,7-dihydroisoquinoline-8(5H)-one (10.0 g, 44.23 mmol) was dissolved in 200 mL of tetrahydrofuran, and (S)-tert-butylsulfinamide (5.9 g, 48.66 mmol) and tetraisopropyl titanate (37.7 g, 132.70 mmol) were added. The mixture was heated to 65 °C under nitrogen protection and stirred for 24 hours.

[0054] After the reaction was complete, 100 mL of water was added to quench the reaction, and the solid was filtered. The filtrate was concentrated, and 100 mL of water was added to the residue. The residue was extracted with ethyl acetate (100 mL × 3 times), and the organic phases were combined. The residue was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane = 1 / 5) to give 13.3 g of (S)-N-(4-bromo-6,7-dihydroisoquinoline-8(5H)-ylidene)-2-methylpropane-2-sulfonamide. [M+H] + =329.12.

[0055] Step E: Synthesis of (S)-N-((R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-yl)-2-methylpropane-2-sulfonamide

[0056] Sodium borohydride (2.3 g, 60.59 mmol) was added in portions to a methanol (400 mL) solution of (S)-N-(4-bromo-6,7-dihydroisoquinoline-8(5H)-ylidene)-2-methylpropane-2-sulfonamide (13.3 g, 40.39 mmol), and the mixture was stirred at -42 °C for 1 hour.

[0057] After the reaction was complete, 100 mL of water was added to quench the reaction, the solvent was evaporated, and 100 mL of water was added to the residue. The mixture was extracted with ethyl acetate (100 mL × 3 times), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 1 / 20) to give 11.1 g of (S)-N-((R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-yl)-2-methylpropane-2-sulfonamide. [M+H] + =331.06. 1 H NMR(400MHz, CDCl3)δ8.58(s,1H),8.57(s,1H),4.59–4.51(m,1H),3.41(d,J=10.0Hz,1H) ,2.83–2.68(m,2H),2.38–2.28(m,1H),2.05–1.95(m,2H),1.94–1.84(m,1H),1.29(s,9H).

[0058] Step F: Synthesis of (R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-amine

[0059] In a solution of (S)-N-((R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-yl)-2-methylpropane-2-sulfonamide (11.1 g, 9.86 mmol) in dichloromethane (100 mL), 40 mL of hydrogen chloride-dioxane solution (4 M) was added, and the mixture was stirred at room temperature for 5 hours.

[0060] After the reaction was complete, the solvent was evaporated from the mixture, and 100 mL of water was added to the residue. The pH was adjusted to 9 with sodium hydroxide solution (1 M), and the mixture was extracted with ethyl acetate (100 mL × 3 times). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 1 / 10) to give 7.2 g of (R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-amine. [M+H] + =227.11.

[0061] Step G: Synthesis of (R)-N-(4-bromo-5,6,7,8-tetrahydroisoquinoline-8-yl)propionamide

[0062] (R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-amine (7.2 g, 31.70 mmol) and triethylamine (8.8 mL, 63.41 mmol) were dissolved in dichloromethane (100 mL), and propionyl chloride (3.1 mL, 34.87 mmol) was added dropwise at 0 °C. The mixture was stirred at room temperature for 5 minutes.

[0063] After the reaction was complete, water was added to the mixture, and the mixture was extracted with dichloromethane (100 mL × 3 times). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 1 / 20) to give 8.5 g of (R)-N-(4-bromo-5,6,7,8-tetrahydroisoquinoline-8-yl)propionamide. [M+H] + =283.12.

[0064] Step H:

[0065] (R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-amine (100 mg, 0.35 mmol) and pinacol ester of 2-cyanoquinoline-6-borate (119 mg, 0.42 mmol) were dissolved in a mixed solvent of 5.0 mL dioxane and 1.0 mL water. Sodium carbonate (76 g, 0.71 mmol) and tetrakis(triphenylphosphine)palladium (8 mg, 0.0071 mmol) were added. The mixture was reacted under nitrogen protection at 85 °C for 6 hours.

[0066] After the reaction was completed, the resulting suspension was filtered, the filter cake was washed with dichloromethane, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 1 / 20) to give 109 mg (R)-N-(4-(2-cyanoquinoline-6-yl)-5,6,7,8-tetrahydroisoquinoline-8-yl)propionamide.

[0067] [M+H] + =357.00. NMR data: 1 HNMR (400MHz, DMSO-d6) δ8.74(d,J=8.5Hz,1H),8.45(s,1H),8.39(s,1H),8.34(d,J=8.4Hz,1H),8.25(d,J=8.7Hz,1H),8.18(d,J=2.0Hz,1H),8.14( d,J=8.4Hz,1H),7.99(dd,J=8.7,2.0Hz,1H),5.15(q,J=6.5Hz,1H),2.71– 2.63(m,2H),2.26–2.12(m,2H),1.99–1.66(m,4H),1.09(t,J=7.6Hz,3H).

[0068] Example 2: Preparation of compound A hydrochloride:

[0069] Weigh 27 mg of hydrochloric acid (concentration 36-38%), add 2 ml of acetone, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection. Dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid with a purity of 99.21%.

[0070] Example 3: Preparation of compound A hydrobromide:

[0071] Weigh 42 mg of hydrobromic acid (concentration 48%), add 2 ml of acetone, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection. Dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid with a purity of 99.01%.

[0072] Example 4: Preparation of compound A sulfate:

[0073] Weigh 25 mg of sulfuric acid, add 2 ml of acetone, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection. Dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid with a purity of 97.61%.

[0074] Example 5: Preparation of compound A phosphate:

[0075] Weigh 25 mg of phosphoric acid, add 2 ml of acetone and 100 μl of water, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection. Dry the filter cake under vacuum at 50 °C for 1 day to obtain a white solid with a purity of 98.95%.

[0076] Example 6: Preparation of compound A benzenesulfonate:

[0077] Weigh 39.5 mg of benzenesulfonic acid, add 2 ml of ethyl acetate, stir, and then add 89 mg of the compound of formula I prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection, and dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid.

[0078] Example 7: Preparation of compound A, p-toluenesulfonate:

[0079] Weigh 47.6 mg of p-toluenesulfonic acid, add 2 ml of acetone, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then add 4 ml of isopropyl ether and continue to react at room temperature for 1 day. Then filter under nitrogen protection, and dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid.

[0080] Example 8: Preparation of compound A maleate:

[0081] Weigh 29.8 mg of maleic acid, add 2 ml of acetone, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then add 4 ml of isopropyl ether and continue to react at room temperature for 1 day. Then filter under nitrogen protection, and dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid.

[0082] Example 9: Preparation of compound A fumarate:

[0083] Weigh 29.8 mg of fumaric acid, add 2 ml of acetone, stir, and then add 89 mg of the compound of formula I prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection, and dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid.

[0084] Example 10: Preparation of compound A oxalate:

[0085] Weigh 22.5 mg of oxalic acid, add 2 ml of acetone, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection. Dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid.

[0086] Example 11: Preparation of compound A succinate:

[0087] Weigh 29.5 mg of succinic acid, add 2 ml of acetone, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection. Dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid.

[0088] Example 12 Preparation of compound A adipate:

[0089] Weigh 36.5 mg of adipic acid, add 2 ml of ethyl acetate, stir, and then add 89 mg of compound A prepared in Example 1. React at room temperature for 1 day, then filter under nitrogen protection. Dry the filter cake under vacuum at 50°C for 1 day to obtain a white solid.

[0090] Example 13 Bioactivity Assessment

[0091] Detection methods

[0092] In this paper, the inventors used the H295R Steroidogenesis Assay System to test the enzyme activities of human CYP11B1, human CYP11B2, etc. The in vitro H295R Steroidogenesis Assay System utilizes the human adrenal cancer cell line (NCI-H295R cells) to construct a level 2 "in vitro assay, providing mechanistic data" for screening and prioritization purposes. The development and standardization of this method were carried out in a multi-step process for screening the chemical effects of steroidogenesis. The H295R assay method has been optimized and validated according to the OECD Test Guideline No. 456 H295R Steroidogenesis Assay.

[0093] Inhibition of aldosterone synthase

[0094] NCI-H295R cells can be purchased from ATCC. After culturing H295R cells from the original ATCC batch, the cells should be cultured for five generations (i.e., the cells divide four times), and then the cells that have been passaged five times should be frozen and stored in liquid nitrogen.

[0095] H295R cells were cultured in a 37°C, 5% CO2 incubator, with the culture medium changed 2-3 times per week. Cells were passaged when they reached approximately 85-90% confluence. The culture medium was then aspirated and replaced with DPBS (calcium-free). 2+ Mg 2+Wash three times, digest with trypsin for 1-3 min, add 3 mL of culture medium to stop digestion and aspirate cells, then wash away any remaining cells with 1 mL of culture medium and combine in a 15 mL centrifuge tube. Centrifuge at 800 rpm for 5 min at room temperature, discard the supernatant, resuspend the pellet in 3 mL of culture medium, and count the cell suspension. Discard the edge wells of a 96-well plate, and seed 50,000 cells per well in the remaining wells. Add 100 μL of 10% FBSDMEM:F12 (1:1) basal medium per well and incubate overnight. Replace with 150 μL of basal medium containing 10 μM Forskolin and incubate for 48 h. After 48 h, replace with basal medium containing 10 μM deoxycorticosterone. Dissolve the compound in DMSO to prepare a 100 mM stock solution. Starting at 100 mM, perform a 3-fold serial dilution in DMSO to obtain 10 concentration points. Ten concentration points were further diluted 10-fold with DMEM:F12 (1:1) blank medium, with an initial concentration of 10 mM. 1.5 μL of each concentration of the compound was added to the cells, with a final DMSO concentration of 0.1% and an initial compound concentration of 100 μM. After incubation for 48 h, 40 μL of cell supernatant was collected, and aldosterone and cortisol levels were analyzed using LCMS.

[0096] Cell viability assay

[0097] After collecting the supernatant, add 100 μL of 10% CCK8 assay reagent to each well, incubate at 37°C for 10 min, mix thoroughly by tapping, and then measure the OD value at 405 nm using a microplate reader. A 70% methanol group was set as a negative control, and DMSO solvent controls were set as a positive control. The %viable cells were calculated using the following formula: %viable cells = (OD cmpd – OD Avg MeOH [=100% dead]) ÷ (OD Avg SCs [=100% viability] – OD Avg MeOH [=100% dead])

[0098] Wells with cell viability below 80% should not be included in the final data analysis. In cases of cytotoxicity approaching 20%, inhibition of steroid production should be carefully evaluated to ensure that cytotoxicity is not the cause of inhibition. Furthermore, data with cell viability exceeding 120% should be labeled to identify potential false positives.

[0099] The inhibition rate was calculated using the following formula: Inhibition rate % = (Peak Area Avg SCs - Peak Area cmpd) / (Peak Area Avg SCs - Peak Area blank) × 100

[0100] Plotting the logarithm of compound concentration on the x-axis and inhibition rate on the y-axis, a nonlinear regression curve was fitted using Graphpad 9.0 to calculate the IC50 value (Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope))), Ki = IC50 / (1 + [S] / Km). The test results are shown in Table 1. Unless otherwise specified, it is assumed that all proteases are competitively inhibited. Selectivity = CYP11B1Ki(nM) / CYP11B2 Ki(nM); where A represents a selectivity value between 0 and 50, B represents a selectivity value between 51 and 100, C represents a selectivity value between 101 and 150, and D represents a selectivity value above 151.

[0101] Table 1. Inhibitory effects of compounds on CYP11B2

[0102] As shown in Table 1, the experimental results of the present invention have a good inhibitory effect on CYP11B2, which is better than that of the control compound Baxdrostat. In addition, the compound of the present invention has excellent selectivity for CYP11B2, which can selectively inhibit CYP11B2 while weakly inhibiting CYP11B1.

[0103] Example 14: Rat Pharmacokinetic Study

[0104] Experimental materials

[0105] SD rats: male, 180-250g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0106] Reagents: DMSO (dimethyl sulfoxide), PEG-400 (polyethylene glycol 400), physiological saline, heparin, acetonitrile, formic acid, and propranolol (internal standard) are all commercially available.

[0107] Instrument: AB SCIEX QTRAP 5500+.

[0108] Experimental methods

[0109] The compound from Example 1 of this invention was dissolved in a DMSO-PEG-400-physiological saline (5:60:35, v / v / v) system. After intravenous or gavage administration to rats, 200 μL of venous blood was collected at 15 min, 30 min, 1 h, 2 h, 5 h, 7 h, and 24 h (an additional 5 min was collected for the IV group) into EDTA-K2 anticoagulant tubes. The tubes were centrifuged at 12000 rpm for 2 min, and the plasma was stored at -80℃ for later analysis. A precise amount of the test sample was dissolved in DMSO to a concentration of 2 mg / mL as a stock solution. An appropriate amount of the compound stock solution was accurately pipetted and diluted with acetonitrile to prepare a series of standard solutions. 10 μL of each of the above standard solutions was accurately pipetted and added to 90 μL of blank plasma. The mixture was vortexed to prepare plasma samples with concentrations equivalent to 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL. Two samples were analyzed for each concentration to establish a standard curve. Take 30 μL of plasma (diluted 5-fold at 5 min, 15 min, and 30 min after intravenous administration), add 150 μL of propranolol (50 ng / mL) in acetonitrile solution, vortex to mix, add 100 μL of purified water, vortex again, centrifuge at 4000 rpm for 5 min, and collect the supernatant for LC-MS analysis. LC-MS detection conditions are as follows:

[0110] Column: YMC Triart C18, 50*3.0mm, 2.1μm.

[0111] Mobile phase: water (0.1% formic acid) - acetonitrile. Gradient elution is performed according to the table below.

[0112] Data processing

[0113] After LC-MS determination of blood drug concentration, pharmacokinetic parameters were calculated using WinNonlin 6.1 software and a non-compartmental model method. The test results are shown in the table.

[0114] Table 2. Pharmacokinetics of the compounds of this invention in rats

[0115] As shown in Table 2, the compounds of this invention exhibit good pharmacokinetic characteristics in SD rats, with C1 values ​​for both intravenous and gavage administration. max and AUC last All were superior to the positive control, with good absorption, high absolute bioavailability, and half-life comparable to or better than the control compound.

[0116] Example 15: Pharmacokinetic Study of Compounds in Crab-Eating Monkeys

[0117] Experimental materials

[0118] Crab-eating macaque: Male, 180-250g, purchased from Guangxi Xiongsen Primate Experimental Animal Breeding and Development Co., Ltd.

[0119] Reagents: DMSO (dimethyl sulfoxide), PEG400, physiological saline, heparin, acetonitrile, formic acid, and propranolol (internal standard) were all commercially available.

[0120] Instrument: AB SCIEX 7500.

[0121] Experimental methods

[0122] The compound was dissolved in a DMSO-PEG-400-physiological saline (5:60:35, v / v / v) system. After administration by gavage to cynomolgus monkeys, 200 μL of venous blood was collected at 30 min, 60 min, 90 min, 2 h, 3 h, 5 h, 8 h, and 24 h in EDTA-K2 anticoagulant tubes. The tubes were centrifuged at 12000 rpm for 2 min, and the plasma was stored at -80℃ for later analysis. A precise amount of the test sample was dissolved in DMSO to a concentration of 2 mg / mL to prepare a stock solution. An appropriate amount of the stock solution was accurately pipetted and diluted with acetonitrile to prepare a series of standard solutions. Accurately pipette 10 μL of each of the above standard series solutions and add 90 μL of blank plasma. Vortex to mix, preparing plasma samples with concentrations equivalent to 0.3, 1, 3, 10, 30, 100, 300, 1000, and 3000 ng / mL, and quality control samples with concentrations of 2.4, 120, and 2400 ng / mL. Perform dual-sample analysis for each concentration and establish a standard curve. Take 30 μL of plasma and add 150 μL of acetonitrile solution containing propranolol (50 ng / mL) as internal standard. Vortex to mix, then add 100 μL of purified water, vortex again, centrifuge at 4000 rpm for 5 min, and analyze the supernatant by LC-MS. The LC-MS detection conditions are as follows:

[0123] Column: YMC Triart C18, 50*3.0mm, 2.1μm.

[0124] Mobile phase: water (0.1% formic acid) - acetonitrile. Gradient elution is performed according to the table below.

[0125] Data processing

[0126] After LC-MS determination of blood drug concentration, pharmacokinetic parameters were calculated using WinNonlin 6.1 software and a non-compartmental model method. The test results are shown in the table.

[0127] Table 3. Pharmacokinetic results of the compounds of this invention in cynomolgus monkeys.

[0128] As shown in Table 3, the series of compounds of this invention all exhibit good pharmacokinetic characteristics in cynomolgus monkeys. The post-oral absorption exposure levels were higher or comparable to the positive control Baxdrostat. max and AUC last All are superior to the positive control, with a better half-life, better absorption, and higher absolute bioavailability.

[0129] Example 16: Pharmacodynamic evaluation of the compounds of the present invention in an animal model of chronic heart failure.

[0130] Experimental plan:

[0131] ZSF1 Rat(Obese) and ZSF1 Rat(Lean) animals were purchased from Nanjing Vital River Pharmaceuticals (aged 13–16 weeks) and fed a special diet (K5008) until 30–33 weeks of age. All animals were housed in a barrier environment, 2–3 animals per cage. ZSF1 Rat(Obese) animals were stratified and randomly assigned to groups based on body weight and aldosterone levels. An Alzet osmotic pump pre-loaded with Saline (0.9% sodium chloride injection) or containing ANGII solution (90 ng / kg / min) was implanted subcutaneously in the back. ANGII was released at a constant rate (0.25 μL / hr) for 30 days during the experiment, and the osmotic pump was removed after the experiment. The rats were orally administered the solvent (0.5% CMC-Na) or compound A once daily for 30 days. Cardiac function parameters of the rats were measured using a small animal ultrasound machine (Vevo 3100lite) before and after the experiment. Left ventricular ejection fraction (LVEF) was calculated using the following formula: LVEF = (EDV - ESV) / ​​EDV * 100%; where LVEF represents the left ventricular ejection fraction; EDV represents the diastolic volume of the left ventricle; and ESV represents the systolic volume of the left ventricle. The results are shown in Figure 1, where "(***, p < 0.001)" in Figure 1 indicates a comparison between ZSF1(Obese)_AngⅡ_Vehicle and ZSF1(Obese)_AngⅡ_compound A.

[0132] Note: PO: Oral administration via gavage; QD: Once daily. Vehicle: 0.5% sodium carboxymethyl cellulose (CMC-Na).

[0133] As can be seen from the results in Figure 1, compound A of the present invention can increase the left ventricular ejection fraction in an animal model of chronic heart failure, and has a significant effect on improving cardiac function.

[0134] It should be understood that the above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Those skilled in the art can make improvements or modifications based on the above description, and all such improvements and modifications should fall within the protection scope of the appended claims of the present invention.

Claims

1. Use of a compound A or a salt thereof in the preparation of a medicament for the prevention or treatment of heart failure (HF)-related diseases, characterized in that, The structural formula of compound A is:

2. The use according to claim 1, characterized in that, The heart failure-related diseases mentioned are selected from: chronic heart failure.

3. The use according to claim 2, characterized in that, The chronic heart failure mentioned is selected from: congestive heart failure (CHF).

4. The use according to claim 2, characterized in that, The chronic heart failure mentioned is selected from heart failure with reduced ejection fraction (HFrEF).

5. The use according to claim 2, characterized in that, The chronic heart failure mentioned is selected from: heart failure with improved ejection fraction (HFimpEF).

6. The use according to claim 2, characterized in that, The chronic heart failure mentioned is selected from: heart failure with mildly reduced ejection fraction (HFmrEF).

7. The use according to claim 2, characterized in that, The chronic heart failure mentioned is selected from heart failure with preserved ejection fraction (HFpEF).

8. The use according to any one of claims 1-7, characterized in that, The frequency of administration of the drug is selected from: once a day, twice a day, three times a day, once every two days, once every three days, and once a week.

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