Medical use of compound a or pharmaceutically acceptable salt thereof

Abstract

The present invention relates to the technical field of chemicals, provides a medical use of a compound A or a pharmaceutically acceptable salt thereof, and specifically relates to a use of the compound A or the pharmaceutically acceptable salt thereof in preparation of a drug for preventing or treating nephropathy-related diseases.

Description

Medical use of a compound A or a pharmaceutically acceptable salt thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of chemical drugs, and provides a medical use of a compound A or a pharmaceutically acceptable salt thereof, and particularly relates to a use of a compound A or a pharmaceutically acceptable salt thereof in preparation of a drug for preventing or treating diseases related to kidney diseases. BACKGROUND

[0002] Aldosterone is a steroid hormone with mineralocorticoid activity. It is mainly produced by the zona glomerulosa of the adrenal cortex in response to angiotensin II, adrenocorticotropic hormone and increased serum potassium content. The main physiological role of aldosterone in the kidney is to maintain sodium and potassium balance by regulating cation exchange (Na+ reabsorption and K+ secretion) in the distal renal unit. However, aldosterone has also been shown to be a pro-inflammatory and pro-fibrotic hormone in blood vessels, heart and kidney. The effects of aldosterone on gene expression are mediated through binding to the mineralocorticoid receptor (MR) and the classic nuclear hormone receptor pathway.

[0003] CYP11B2 (aldosterone synthase) is mainly expressed in the adrenal cortical glomerular zone, and is known as an enzyme that catalyzes a series of reactions from 11-deoxycorticosterone (i.e., aldosterone precursor) to aldosterone. The finding that aldosterone is associated with organ dysfunction has attracted attention. It has been reported that CYP11B2 inhibitors can inhibit the production of aldosterone in studies using enzymes and cultured cells, and have an inhibitory effect on the production of aldosterone and a therapeutic effect in studies using various experimental animal models. In addition, it has been confirmed that CYP11B2 inhibitors exhibit an effect of lowering the level of aldosterone in plasma and urine and a blood pressure lowering effect in patients with hypertension and primary aldosteronism. Finding a means to block the biosynthetic pathway of aldosterone is a highly achievable approach for establishing an effective treatment method for various diseases associated with aldosterone.

[0004] Chronic kidney disease (CKD) is the leading cause of kidney damage and end-stage renal disease (ESRD). The 5-year survival rate of dialysis patients is 35%, which decreases to only 25% in diabetic dialysis patients. Therefore, CKD places a heavy burden on global healthcare systems, for example, in the United States, the cost per patient per year exceeds 75,000 dollars. In addition to the direct impact on the kidney, reduced kidney function is also a major cause of cardiovascular events. Aldosterone, as an important component of the renin-angiotensin-aldosterone system (RAAS), promotes inflammation and fibrosis in the kidney and blood vessels. It has been reported that elevated aldosterone is negatively correlated with eGFR, positively correlated with 24h urinary protein at baseline, and independently associated with the progression of CKD and new-onset ESKD.

[0005] Aldosterone synthase (CYP11B2) is a key enzyme in the biosynthesis of aldosterone, by selectively inhibiting the biosynthesis of aldosterone. Reducing aldosterone can play a kidney protective effect in chronic kidney disease (CKD) and treat related diseases.

[0006] PCT / CN2024 / 142727 discloses an aldosterone synthase inhibitor and its preparation method and use, and discloses the use of an aldosterone synthase inhibitor in a drug for preventing or treating diseases related to CYP11B2, and seeks an aldosterone synthase inhibitor small molecule compound with excellent effect for treating chronic kidney disease, which has become a problem to be solved in the art. SUMMARY

[0007] In view of the problems existing in the prior art, the present application provides a medical use of compound A or a pharmaceutically acceptable salt thereof to solve the problems existing in the prior art.

[0008] The present application is realized by the following technical solutions:

[0009] The present application provides the use of compound A or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing or treating diseases related to kidney disease, characterized in that the structural formula of the compound A is:

[0010] Further, as a preferred technical solution of the present application, the kidney disease related disease is selected from chronic kidney disease (CKD).

[0011] Further, as a preferred technical solution of the present application, the kidney disease related disease or chronic kidney disease is selected from diabetic nephropathy (DKD).

[0012] Further, as a preferred technical solution of the present application, the kidney disease related disease or chronic kidney disease is selected from hypertensive nephropathy, lupus nephritis (LN), allergic purpura nephritis, hepatitis B virus related nephritis, obesity related nephropathy or ANCA related vasculitis.

[0013] Further, as a preferred technical solution of the present application, the kidney disease related disease or chronic kidney disease is selected from IgA nephropathy, IgG nephropathy, IgM nephropathy, membranous nephropathy (MN), minimal change nephropathy (MCD) or focal segmental glomerulosclerosis (FSGS).

[0014] Further, as a preferred technical solution of the present application, the kidney disease related disease or chronic kidney disease is selected from glomerulonephritis and glomerulosclerosis.

[0015] Further, as a preferred technical solution of the present application, the renal disease related disease or chronic kidney disease is selected from: Alport syndrome, thin basement membrane nephropathy or polycystic kidney disease (ADPKD).

[0016] Further, as a preferred technical solution of the present application, the renal disease related disease or chronic kidney disease is selected from: renal tubulointerstitial nephritis.

[0017] Further, as a preferred technical solution of the present application, the pharmaceutically acceptable salt is selected from hydrochloride, hydrobromide, phosphate, nitrate, sulfate, acetate, propionate, malonate, succinate, valerate, glutarate, adipate, oxalate, L-proline, lactobionate, glycine, alanine, arginine, lactic acid, cinnamic acid, fumaric acid, mandelic acid, maleic acid, hippuric acid, tartaric acid, citric acid, malic acid, succinic acid, 2-naphthalenesulfonic acid, 1,5-naphthalenedisulfonic acid, camphorsulfonic acid, benzoic acid, salicylic acid, benzenesulfonic acid, methanesulfonic acid or p-toluenesulfonic acid; preferably, the pharmaceutically acceptable salt is independently selected from hydrochloride, hydrobromide, phosphate, sulfate, benzenesulfonic acid, p-toluenesulfonic acid, maleic acid, fumaric acid, oxalic acid, succinic acid or adipic acid.

[0018] Further, as a preferred technical solution of the present application, the molar ratio of compound A to acid molecule in the pharmaceutically acceptable salt is 1:0.3-1:3.5; preferably, the molar ratio of compound A to acid molecule is 1:1, 1:2, 1:3, 2:1, 3:1; more preferably, the molar ratio of compound A to acid molecule is 1:1;

[0019] Further, as a preferred technical solution of the present application, the use amount of compound A or its pharmaceutically acceptable salt (all based on compound A) in the medicine is 0.25mg-50mg; preferably, the use amount of compound A or its pharmaceutically acceptable salt (all based on compound A) in the medicine is 0.25mg-25mg; more preferably, the use amount of compound A or its pharmaceutically acceptable salt (all based on compound A) in the medicine is 0.25mg-10mg; most preferably, the use amount of compound A or its pharmaceutically acceptable salt (all based on compound A) in the medicine is 0.25mg, 0.5mg, 0.75mg, 1mg, 2mg, 3mg, 4mg, 6mg, 7mg, 8mg, 9mg or 10mg.

[0020] Further, as a preferred technical solution of the present application, the compound A or its pharmaceutically acceptable salt is a crystal form, amorphous or a mixture thereof.

[0021] Further, as a preferred technical solution of the present application, one or more hydrogen atoms on the compound A or its pharmaceutically acceptable salt are substituted by isotope deuterium.

[0022] Further, as a preferred technical solution of the present application, the administration frequency of the drug is selected from the group consisting of once a day, twice a day, three times a day, once every two days, once every three days, once a week. Preferably, the administration frequency of the drug is once a day.

[0023] Further, as a preferred technical solution of the present application, the present application also provides a method for preventing and / or treating a kidney disease related disease, comprising the following steps: administering a therapeutically effective amount of the compound A or its pharmaceutically acceptable salt of the present application to a patient in need thereof. Preferably, the kidney disease related disease is selected from the group consisting of chronic kidney disease (CKD), diabetic kidney disease (DKD), hypertensive nephropathy, lupus nephritis (LN), Henoch-Schonlein purpura nephritis, hepatitis B virus associated nephritis, obesity associated nephropathy, ANCA associated vasculitis, IgA nephropathy, IgG nephropathy, IgM nephropathy, membranous nephropathy (MN), minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), glomerulonephritis, glomerulosclerosis, Alport syndrome, thin basement membrane nephropathy or polycystic kidney disease (ADPKD), tubulointerstitial nephritis.

[0024] Further, the compound A of the present application can significantly reduce proteinuria in patients with hypertensive nephropathy or chronic kidney disease, preferably the compound A of the present application can significantly reduce urinary albumin creatinine ratio (UACR) in patients with hypertensive nephropathy or chronic kidney disease.

[0025] For the sake of clarity, the general terms used in the description of the compounds are defined herein.

[0026] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as being indefinite or unclear if it is not specifically defined, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding product or active ingredient thereof. The term "pharmaceutically acceptable" used herein refers to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0027] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present application, prepared from a compound of the present application having specific substituents discovered by the present application with a pharmaceutically acceptable acid or base.

[0028] In addition to salt forms, the compounds provided herein can also take prodrug forms. Prodrugs of the compounds described herein are readily converted by chemical or physiological processes, once inside the body, to provide the compounds of the present application. The term "prodrug" is intended to represent a compound that is a drug in itself, or a drug candidate, and is designed to undergo some

[0029] Certain compounds of the present application can exist in unsolvated as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present application.

[0030] The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as the racemic mixtures and other mixtures thereof, such as pure or enriched mixtures of enantiomers or stereoisomers, all of which are intended to be within the scope of the present application. Additional asymmetric carbon atoms can be present in a substituent group. All such isomers, as well as mixtures thereof, are included within the scope of the present application.

[0031] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or by chiral reagents or other conventional techniques. If desired, one enantiomer of a compound of the present application can be obtained by asymmetric synthesis or derivatization with a chiral auxiliary, separation of the resulting diastereomeric mixture, and cleavage of the auxiliary to provide the pure desired enantiomer. Alternatively, when a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group) is present in the molecule, diastereomeric salts with an appropriate optically active acid or base can be formed, and the pure enantiomer recovered by conventional means well known in the art, followed by recovery. In addition, separation of the enantiomers and diastereomers is typically accomplished by chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamates from amines).

[0032] The atoms of the molecules of the compounds of the present application are isotopically enriched. Isotopic derivatization generally results in an increase in half-life, a decrease in clearance, metabolic stability, and an increase in in vivo activity. Also included is an embodiment wherein at least one atom is replaced by an atom having the same atomic number (number of protons) and a different mass number (number of protons and neutrons). Examples of isotopes that are included in the compounds of the present application include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, which respectively include 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 which emit radiation as they decay, such as 3 H or 14 C, can be used in the manufacture of pharmaceuticals or in the localised anatomical examination of compounds in vivo. Stable isotopes neither decay nor change in amount as they are used, nor are they radioactive, and so they can be used safely. When the atoms which make up the molecules of the compounds of the application are isotopes, isotopic compounds can be converted according to the general methods by substituting the reagents used in the synthesis with reagents containing the corresponding isotopes.

[0033] The compounds of the present application can contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be specially labeled for use as molecular probes, such as with radioactive isotopes (e.g., 2 H), iodine-125 125 I), or carbon-14 14 C). All isotopic variations of the compounds of the present application, whether radioactive or not, are encompassed within the scope of the present application.

[0034] Further, the compounds of the present application wherein one or more hydrogen atoms are replaced by deuterium (2H) are provided. The deuterium substituted compounds of the present application have the effect of increasing the half-life, decreasing the clearance rate, increasing the metabolic stability, and increasing the in vivo activity of the compounds.

[0035] The methods for preparing the isotopic derivatives generally include phase transfer catalysis. For example, a preferred method of deuteriation employs a phase transfer catalyst (e.g., a tetraalkylammonium salt, NBu4HSO4). The use of a phase transfer catalyst to exchange the methylene protons of diphenylmethane compounds results in the introduction of higher deuterium than is achieved by reduction with deuterated silane (e.g., triethyldeuteratedsilane) or with sodium borodeuteride in the presence of an acid (e.g., methanesulfonic acid) or with Lewis acids such as aluminum trichloride.

[0036] The term "pharmaceutically acceptable carrier" means any formulation carrier or medium that does not interfere with the biological activity of the active substance of the application and that is nontoxic to the host or patient in whom it is administered. Representative carriers include water, oil, vegetable and mineral, cream bases, lotion bases, ointment bases, and the like. These bases include suspending agents, viscosity increasing agents, penetration enhancers, and the like. Their formulation is well known to those skilled in the art of cosmetics or topical pharmaceuticals. Additional information on carriers can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0037] The term "excipient" generally refers to a carrier, diluent, and / or vehicle needed to formulate an effective pharmaceutical composition.

[0038] The term "effective amount" or "therapeutically effective amount" with respect to a pharmaceutical or pharmacological agent means a sufficient amount of the agent to achieve the intended effect without being toxic to the subject. With respect to the oral dosage forms of the application, an "effective amount" of one active substance in a composition means the amount needed to achieve the intended effect in conjunction with another active substance in the composition. The determination of an effective amount will vary from subject to subject, depending on the age and general condition of the subject, as well as the particular active substance, and an appropriate effective amount for a given case can be determined by those skilled in the art based on routine testing.

[0039] The term "active ingredient," "therapeutic agent," "active substance," or "active agent" means a chemical entity that is effective in treating a target disorder, disease, or condition.

[0040] "Optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. BRIEF DESCRIPTION OF DRAWINGS

[0041] 1) Figure 1 shows glomerulosclerosis score (50 glomerulosclerosis total score) in a chronic kidney disease animal model for compounds of the application (glomerulosclerosis score comparison ZSFl (Obese)_Ang II_Vehicle and ZSFl (Obese)_Ang II_Compound A (*, p<0.05));

[0042] 2) Figure 2 shows kidney H&E staining pictures in a chronic kidney disease animal model for compounds of the application;

[0043] 3) Figure 3 shows the change in urine microalbumin to urine creatinine ratio on day 3 of the second phase of dose escalation (1 mg / kg) in the spontaneously hypertensive monkey model compared to pre-dose. DETAILED DESCRIPTION

[0044] The application will be further described in conjunction with the following examples, but the scope of the application is not limited to the examples.

[0045] Example 1

[0046] Synthesis of (R)-N-(4-(2-cyanoquinolin-6-yl)-5,6,7,8-tetrahydroisoquinolin-8- yl)propanamide

[0047] Specific operation steps:

[0048] Step A: Synthesis of ethyl 5-bromo-4-methylnicotinate

[0049] Ethyl 5-bromo-4-methylnicotinate (54.6 g, 223.69 mmol) and methyl acrylate (48.1 g, 559.22 mmol) were dissolved in 500 mL of tetrahydrofuran, and LDA (123 mL, 246.06 mmol, 2M) was added dropwise at -78°C. The mixture was stirred for 30 minutes, and then the reaction was continued at -78°C for 2 hours.

[0050] After the reaction was completed, the mixture was filtered, and water was added to the filtrate. The mixture was extracted with ethyl acetate (300 mL x 3 times), and the organic phase was combined, washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The obtained residue was purified by column chromatography on silica gel (eluent: ethyl acetate / n-hexane = 1 / 10) to obtain 54.6 g of ethyl 5-bromo-4-methylnicotinate. [M+H] + = 244.05.

[0051] Step B: Synthesis of methyl 4-bromo-8-oxo-5,6,7,8-tetrahydroisoquinoline-7- carboxylate

[0052] Ethyl 5-bromo-4-methylnicotinate (54.6 g, 223.69 mmol) and methyl acrylate (48.1 g, 559.22 mmol) were dissolved in 800 mL of tetrahydrofuran, and LDA (123 mL, 246.06mmol, 2M) was added dropwise at -78°C. The mixture was stirred at -78°C for 30 minutes, and then the reaction was continued at -78°C for 2 hours. After the reaction was completed, the mixture was filtered, and water was added to the filtrate. Then, the mixture was extracted with ethyl acetate (300 mL x 3 times), and the organic layer was combined, washed with saturated brine (500 mL), dried over anhydrous sodium

[0053] After the reaction was completed, 400 mL of 10% acetic acid aqueous solution was added to quench the reaction, the organic solvent was removed by distillation, and the mixture was extracted with ethyl acetate (300 mL x 3 times), the organic phase was combined, washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane = 1 / 10) to obtain 31.5 g of methyl 4-bromo-8-oxo-5,6,7,8-tetrahydroisoquinoline-7-carboxylate. [M+H] + = 284.06.

[0054] Step C: Synthesis of 4-bromo-6,7-dihydroisoquinolin-8(5H)-one

[0055] Methyl 4-bromo-8-oxo-5,6,7,8-tetrahydroisoquinoline-7-carboxylate (31.5 g, 110.87 mmol) was dissolved in 300 mL of hydrochloric acid (6M), and the mixture was stirred at 105°C under reflux for 16 hours.

[0056] After the reaction was completed, the solvent was removed by distillation, 300 mL of water was added, the pH was adjusted to ~9 with 1N sodium hydroxide aqueous solution, the mixture was extracted with ethyl acetate (200 mL x 3 times), the organic phase was combined, washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane = 1 / 8) to obtain 19.6 g of 4-bromo-6,7-dihydroisoquinolin-8(5H)-one. [M+H] + = 226.05.

[0057] Step D: Synthesis of (S)-N-(4-bromo-6,7-dihydroisoquinolin-8(5H)-ylidene)-2-methylpropane-2-sulfonamide

[0058] 4-bromo-6,7-dihydroisoquinolin-8(5H)-one (10.0 g, 44.23 mmol) was dissolved in 200 mL of tetrahydrofuran, (S)-tert-butylsulfinamide (5.9 g, 48.66 mmol) and tetraisopropyl titanate (37.7 g, 132.70 mmol) were added, and the mixture was stirred at 65°C under nitrogen protection for 24 hours.

[0059] 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.

[0060] Step E: Synthesis of (S)-N-((R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-yl)-2-methylpropane-2-sulfonamide

[0061] 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.

[0062] 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).

[0063] Step F: Synthesis of (R)-4-bromo-5,6,7,8-tetrahydroisoquinoline-8-amine

[0064] 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.

[0065] 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.

[0066] Step G: Synthesis of (R)-N-(4-bromo-5,6,7,8-tetrahydroisoquinoline-8-yl)propionamide

[0067] (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.

[0068] 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.

[0069] Step H:

[0070] (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.

[0071] 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.

[0072] [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).

[0073] Example 2: Preparation of compound A hydrochloride:

[0074] 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%.

[0075] Example 3: Preparation of compound A hydrobromide:

[0076] 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%.

[0077] Example 4: Preparation of compound A sulfate:

[0078] 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%.

[0079] Example 5: Preparation of compound A phosphate:

[0080] 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%.

[0081] Example 6: Preparation of compound A benzenesulfonate:

[0082] 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.

[0083] Example 7: Preparation of compound A, p-toluenesulfonate:

[0084] 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.

[0085] Example 8: Preparation of compound A maleate:

[0086] 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.

[0087] Example 9: Preparation of compound A fumarate:

[0088] 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.

[0089] Example 10: Preparation of compound A oxalate:

[0090] 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.

[0091] Example 11: Preparation of compound A succinate:

[0092] 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.

[0093] Example 12 Preparation of compound A adipate:

[0094] 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.

[0095] Example 13 Bioactivity Assessment

[0096] Detection methods

[0097] 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.

[0098] Inhibition of aldosterone synthase

[0099] 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.

[0100] 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.

[0101] Cell viability assay

[0102] 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])

[0103] 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.

[0104] 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

[0105] 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.

[0106] Table 1. Inhibitory effects of compounds on CYP11B2

[0107] 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.

[0108] Example 14: Rat Pharmacokinetic Study

[0109] Experimental materials

[0110] SD rats: male, 180-250g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0111] Reagents: DMSO (dimethyl sulfoxide), PEG-400 (polyethylene glycol 400), physiological saline, heparin, acetonitrile, formic acid, and propranolol (internal standard) are all commercially available.

[0112] Instrument: AB SCIEX QTRAP 5500+.

[0113] Experimental methods

[0114] 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:

[0115] Column: YMC Triart C18, 50*3.0mm, 2.1μm.

[0116] Mobile phase: water (0.1% formic acid) - acetonitrile. Gradient elution is performed according to the table below.

[0117] Data processing

[0118] After LC-MS was used to detect the blood drug concentration, the pharmacokinetic parameters were calculated using WinNonlin 6.1 software and the non-compartmental model method. The test results are shown in Table 2.

[0119] Table 2. Pharmacokinetics of the compounds of this invention in rats

[0120] 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.

[0121] Example 15: Pharmacokinetic Study of Compounds in Crab-Eating Monkeys

[0122] Experimental materials

[0123] Crab-eating macaque: Male, 180-250g, purchased from Guangxi Xiongsen Primate Experimental Animal Breeding and Development Co., Ltd.

[0124] Reagents: DMSO (dimethyl sulfoxide), PEG400, physiological saline, heparin, acetonitrile, formic acid, and propranolol (internal standard) were all commercially available.

[0125] Instrument: AB SCIEX 7500.

[0126] Experimental methods

[0127] 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:

[0128] Column: YMC Triart C18, 50*3.0mm, 2.1μm.

[0129] Mobile phase: water (0.1% formic acid) - acetonitrile. Gradient elution is performed according to the table below.

[0130] Data processing

[0131] 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.

[0132] Table 3. Pharmacokinetic results of the compounds of this invention in cynomolgus monkeys.

[0133] 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.

[0134] Example 16: Pharmacodynamic evaluation of the compounds of the present invention in an animal model of chronic kidney disease.

[0135] Experimental plan:

[0136] ZSF1 Rat(Obese) and ZSF1 Rat(Lean) animals were purchased from Nanjing Vital River Pharmaceutical Co., Ltd. (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 grouped according to body weight and urinary microalbumin levels. A pre-loaded Alzet osmotic pump containing Saline (0.9% sodium chloride injection) or 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 animal was orally administered the solvent (0.5% CMC-Na) or compound A once daily for 30 days. After the experiment, the animals were anesthetized with salbutamol and xylazine, and the kidneys were fixed in 10% formalin and then stained with hematoxylin and eosin (HE) or acetaminophen (PAS). Histologically stained images were statistically analyzed after being graded according to pathological criteria. The experimental results are shown in Figures 1 and 2.

[0137] Note: PO: Oral administration via gavage; QD: Once daily. Vehicle: 0.5% sodium carboxymethyl cellulose (CMC-Na).

[0138] As can be seen from the results in Figures 1 and 2, the kidney pathological sections of compound A of the present invention showed obvious kidney pathological changes and could significantly improve kidney lesions. Among them, the scores of glomerular sclerosis, glomerular vacuolar degeneration, basement membrane thickening, and renal tubular dilation were significantly lower than those of the solvent control group.

[0139] Example 17: The effect of the compound of the present invention on the improvement of kidney disease in a spontaneously hypertensive monkey model.

[0140] Experimental plan:

[0141] This experiment used cynomolgus monkeys with spontaneous hypertension and increased urinary protein, divided into two groups. The efficacy of the compound was explored through a dose-escalation experimental design. Compound A was first administered at 0.2 mg / kg for 7 days, followed by 1 mg / kg for 3 days. On the third day after the 1 mg / kg administration, urinary microalbumin and creatinine levels were measured to investigate whether it improved renal function. The results are shown in Figure 3. In Figure 3, UACR represents the ratio of urinary microalbumin to creatinine, and "(*, p<0.05)" indicates a comparison between the Vehicle group and compound A.

[0142] Note: PO: Oral administration via gavage; QD: Once daily. Vehicle: 0.5% sodium carboxymethyl cellulose (CMC-Na).

[0143] As shown in Figure 3, in the spontaneously hypertensive monkey model with increased urinary protein, the ratio of urinary microalbumin to creatinine in the model group increased compared to before drug administration, while the ratio of urinary microalbumin to creatinine decreased after day 10 of drug administration for compound A of this invention. The rate of change in the ratio of urinary microalbumin to creatinine in the drug administration group was significantly lower than that in the model group. Compound A of this invention has an ameliorative effect on nephropathy.

[0144] 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 Compound A or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the prevention or treatment of a disease associated with kidney disease, characterized in that, The structural formula of the compound A is:

2. Use according to claim 1, characterized in that, The kidney disease-related condition is selected from the group consisting of chronic kidney disease (CKD).

3. Use according to claim 1 or 2, characterized in that, The kidney disease-related condition or chronic kidney disease is selected from the group consisting of diabetic kidney disease (DKD).

4. Use according to claim 1 or 2, characterized in that, The kidney disease-related condition or chronic kidney disease is selected from the group consisting of hypertensive nephropathy, lupus nephritis (LN), Henoch-Schonlein purpura nephritis, hepatitis B virus-associated nephritis, obesity-associated nephropathy, or ANCA-associated vasculitis.

5. Use according to claim 1 or 2, characterized in that, The kidney disease-related condition or chronic kidney disease is selected from the group consisting of IgA nephropathy, IgG nephropathy, IgM nephropathy, membranous nephropathy (MN), minimal change disease (MCD), or focal segmental glomerulosclerosis (FSGS).

6. Use according to claim 1 or 2, characterized in that, The kidney disease-related condition or chronic kidney disease is selected from the group consisting of glomerulonephritis, glomerulosclerosis.

7. Use according to claim 1 or 2, characterized in that, The kidney disease-related condition or chronic kidney disease is selected from the group consisting of Alport syndrome, thin basement membrane nephropathy, or autosomal dominant polycystic kidney disease (ADPKD).

8. Use according to claim 1 or 2, characterized in that, The kidney disease-related condition or chronic kidney disease is selected from the group consisting of tubulointerstitial nephritis.

9. Use according to any one of claims 1 to 8, characterized in that, The drug is administered at a frequency selected from the group consisting of once a day, twice a day, three times a day, once every two days, once every three days, once a week.

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