Use of aromatic biological antagonist in preparing medicament for treating or preventing kidney disease

Abstract

The present invention relates to use of an aromatic biological antagonist in preparing a medicament for treating or preventing a kidney disease. Specifically, the present invention provides use of 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'- biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof in preparing a medicament for treating or preventing a kidney disease, especially in preparing a medicament for treating or preventing focal segmental glomerulosclerosis and immunoglobulin A nephropathy. The compound or the pharmaceutically acceptable salt thereof shows excellent therapeutic effects on focal segmental glomerulosclerosis and immunoglobulin A nephropathy, and has great clinical application prospects.

Description

Use of aromatic ring-containing biological antagonists in the preparation of drugs for treating or preventing kidney diseases TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and relates to use of aromatic ring-containing biological antagonists in the preparation of drugs for treating or preventing kidney diseases. BACKGROUND

[0002] Immunoglobulin A nephropathy (IgAN) is the most common glomerular disease worldwide, and its renal biopsy pathological features are IgA or IgA-based immune complex deposition in the mesangial area of the glomerulus, and the basic histological change is mainly mesangial proliferation. The clinical manifestations are diverse, mainly manifested as hematuria, and can be accompanied by different degrees of proteinuria, hypertension and impaired renal function. IgAN is affected by genetic and environmental factors, and the regional distribution difference is obvious. The incidence of Asians is higher than that of whites, and it is rare in blacks. Current data in China shows that IgAN accounts for about 50% of primary glomerular diseases. The long-term prognosis of IgAN is poor, and 30%-40% of patients will progress to end-stage renal disease (ESRD) within 10-25 years. The risk of progression to ESRD is higher in Asian populations, and is the most common cause of ESRD. The 10, 15 and 20-year kidney survival rates of IgAN in Chinese population are 83%, 74% and 64%, respectively.

[0003] The pathogenesis of IgAN has not been fully elucidated. It is currently believed that IgA1 in the body of an IgAN patient is more prone to self-aggregation or recognition by IgG / IgA to form immune complexes due to glycosylation defects in the hinge region O-glycan, and the immune complexes are deposited in the mesangial region of the glomerulus to further trigger inflammatory and fibrotic responses of the body, damage the glomerulus, and eventually lead to progressive decline of renal function. The renin-angiotensin system (RAS) is in an activated state in IgAN patients, and even in early IgAN patients with normal blood pressure, activation can be detected. The key active factor of RAS, angiotensin II (AngII), can increase blood pressure, glomerular capillary pressure and filtration membrane permeability through hemodynamic effects. In recent years, it has also been found that AngII has multiple non-hemodynamic effects, such as up-regulating the pro-fibrotic factor-β (TGF-β) to aggravate renal fibrosis, promoting mesangial cell proliferation, inducing chemokines to activate macrophages, and increasing extracellular matrix (ECM) deposition. The sustained abnormal activation of RAS eventually leads to symptoms such as hypertension, proteinuria, and loss of renal function in IgAN patients. Endothelin-1 (ET-1) is one of the strongest endogenous vasoconstrictors, and in addition to directly constricting blood vessels to increase blood pressure, it is also a bypass activator of Ang II. In recent years, it has also been found that ET-1 participates in the pathophysiological progression of IgAN by activating endothelin A receptor (ETA), including podocyte dysfunction, renal tubular damage, inflammation and fibrosis. It has also been found in clinical practice that the level of ET-1 in the peripheral blood of IgAN patients is significantly increased and is related to the severity of proteinuria. Therefore, RAS and ET-1 are both key factors in the development of IgAN, and they promote each other.

[0004] In view of the unmet treatment needs of IgAN patients, 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide can specifically bind to AT1 and ETA receptors, and is a highly selective oral small molecule dual-target inhibitor. This compound has strong endothelin 1 and angiotensin II related signal pathway inhibition activity, and has good effects on improving proteinuria and renal pathological damage in IgAN rat models induced by triplets (BSA, SEB, CCl4), and excellent efficacy and safety in IgAN patients, and is expected to better meet the clinical needs of IgAN and reduce the economic burden of social diseases. SUMMARY

[0005] The present application provides a use of a 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a kidney disease.

[0006] In certain embodiments of the present application, the kidney disease is selected from the group consisting of focal segmental glomerulosclerosis and immunoglobulin A nephropathy.

[0007] In certain embodiments of the present application, the medicament is a pharmaceutical composition of a 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof, wherein the dosage or strength of the medicament is 100-800 mg.

[0008] In preferred embodiments of the present application, the dosage or strength of the medicament of a 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof pharmaceutical composition is 100 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 600 mg or 800 mg.

[0009] In preferred embodiments of the present application, the dosage or strength of the medicament of a 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof pharmaceutical composition is 200 mg or 400 mg.

[0010] The present application also provides a method for treating or preventing a kidney disease using a 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof.

[0011] In certain embodiments of the present application, the kidney disease is selected from the group consisting of focal segmental glomerulosclerosis and immunoglobulin A nephropathy.

[0012] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-l,3-diazaspiro[4.4]non-l- en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[l,l'-biphenyl]-2- sulfonamide compound has the structure:

[0013] In preferred embodiments of the application, the pharmaceutically acceptable salt of the 4'-((2- butyl-4-oxo-l,3-diazaspiro[4.4]non-l-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)- 2'-(ethoxymethyl)-[l,l'-biphenyl]-2-sulfonamide compound is selected from the group consisting of a hydrochloride salt, a sulfate salt, a nitrate salt, a hydrobromide salt, a hydrofluoride salt, a hydroiodide salt, a phosphate salt, a 2,5-dihydroxybenzoic acid salt, a l-hydroxy-2-naphthoic acid salt, an acetate salt, a dichloroacetate salt, a trichloroacetate salt, an acetyloxymethoxy acid salt, an adipate salt, a benzenesulfonate salt, a 4-chlorobenzenesulfonate salt, a benzoate salt, a 4-acetamidobenzoate salt, a 4-aminobenzoate salt, a decanoate salt, a hexanoate salt, an octanoate salt, a cinnamate salt, a citrate salt, a cyclohexanesulfamic acid salt, a camphorsulfonic acid salt, an aspartate salt, a camphoric acid salt, a gluconate salt, a glucaronate salt, a glutamate salt, an erythorbate salt, a lactate salt, a malate salt, a mandelate salt, a pyroglutamate salt, a tartrate salt, a dodecylsulfate salt, a dibenzoyl tartrate salt, an ethane- 1,2-disulfonate salt, an ethanesulfonate salt, a formate salt, a fumarate salt, a galacturonate salt, a gentisate salt, a glutarate salt, a 2-ketoglutarate salt, a glycolate salt, a hippurate salt, a isethionate salt, a lactobionate salt, an ascorbate salt, an aspartate salt, a laurate salt, a camphorate salt, a maleate salt, a malonate salt, a mesylate salt, a 1,5-naphthalene disulfonate salt, a naphthalene-2-sulfonate salt, a nicotinate salt, an oleate salt, an orotate salt, an oxalate salt, a palmitate salt, a pamoate salt, a propionate salt, a salicylate salt, a 4-aminosalicylate salt, a sebacate salt, a stearate salt, a succinate salt, a thiocyanate salt, a pantothenate salt, a formate salt, an undecylenate salt, a trifluoroacetate salt, a benzene sulfonate salt, a p-toluene sulfonate salt, or an L-malate salt.

[0014] In further preferred embodiments of the application, the pharmaceutically acceptable salt of the 4'-((2-butyl-4-oxo-l,3-diazaspiro[4.4]non-l-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)- 2'-(ethoxymethyl)-[l,l'-biphenyl]-2-sulfonamide compound is selected from the group consisting of a mesylate salt, a benzene sulfonate salt, an isethionate salt, an ethanesulfonate salt, a hydrochloride salt, a sulfate salt, a p-toluene sulfonate salt, a phosphate salt, or a hydrobromide salt.

[0015] In further preferred embodiments of the application, the pharmaceutically acceptable salt of the 4'-((2-butyl-4-oxo-l,3-diazaspiro[4.4]non-l-en-3-yl)methyl-d2)-N-(4-chloro-5- methylisoxazol-3-yl)-2'-(ethoxymethyl)-[l,l'-biphenyl]-2-sulfonamide compound is the hydrochloride salt.

[0016] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-l,3-diazaspiro[4.4]non-l-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[l,l'- biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof can be administered once a day, twice a day, or three times a day.

[0017] In preferred embodiments of the application, the 4'-((2-butyl-4-oxo-l,3-diazaspiro[4.4]non-l-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[l,l'- biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is the free base.

[0018] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-l,3-diazaspiro[4.4]non-l-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[l,l'- biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is administered in a dose or single dose amount by weight of the free base.

[0019] In certain embodiments of the application, the single dose amount of the 4'-((2-butyl-4-oxo-l,3-diazaspiro[4.4]non-l-en-3-yl)methyl-d2)-N-(4-chloro-5- methylisoxazol-3-yl)-2'-(ethoxymethyl)-[l,l'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is selected from the range of 1 to 1500 mg, preferably 10 to 1200 mg, 20 to 1100 mg, 30 to 1000 mg, 40 to 900 mg, 50 to 800 mg, 60 to 750 mg, 70 to 700 mg, 80 to 650 mg, 90 to 600 mg, 100 to 550 mg, 150 to 500 mg, 200 to 450 mg, 250 to 400 mg, 200 to 400 mg, or 300 to 350 mg; more preferably 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 95 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg.

[0020] In certain embodiments of the application, the single dose of 4'-((2-butyl-4-oxo- 1,3-diazaspiro[4.4]non-1 -en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)- [1,1 '-biphenyl]-2-sulfonamide or a pharmaceutically acceptable salt thereof is 100 mg, 200 mg or 400 mg.

[0021] In certain embodiments of the application, the single dose of 4'-((2-butyl-4-oxo- 1,3-diazaspiro[4.4]non-1 -en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)- [1,1 '-biphenyl]-2-sulfonamide or a pharmaceutically acceptable salt thereof is in the range of 1-1500 mg, and the frequency of administration can be once a day, twice a day or three times a day. Exemplary doses are selected from the group consisting of 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 82.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg, 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg or 1000 mg.

[0022] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1 - en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1 '-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof is administered in a single dose amount in the range of 100-800 mg, with a dosing frequency of once a day, twice a day or three times a day.

[0023] In further preferred embodiments of the application, the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1 - en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1 '-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof is administered in a single dose amount of 200 mg or 400 mg, once a day or twice a day.

[0024] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1 - en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1 '-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof is administered in a single dose amount of 200 mg, once or twice a day, for 14-30 days, followed by an increase in dose to 400 mg, once or twice a day.

[0025] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1 - en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1 '-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof is administered in a single dose amount of 200 mg, once or twice a day, for 14 days, followed by an increase in dose to 400 mg, once or twice a day.

[0026] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1 - en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1 '-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof is administered in a single dose amount of 200 mg, once a day, for 14 days, followed by an increase in dose to 400 mg, once a day.

[0027] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3- diazaspiro[4.4]non-1 -ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-( ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is administered at a dose of 200 mg twice daily, and the dose is increased to 400 mg twice daily after 14 days of continuous administration.

[0028] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3- diazaspiro[4.4]non-1 -ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-( ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is administered at a dose of 200 mg twice daily, and the dose is increased to 400 mg twice daily after 14 days of continuous administration.

[0029] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3- diazaspiro[4.4]non-1 -ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-( ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is administered at a dose of 200 mg-800 mg per day.

[0030] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3- diazaspiro[4.4]non-1 -ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-( ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is administered at a dose of 200 mg-800 mg per day.

[0031] In certain embodiments of the application, the 4'-((2-butyl-4-oxo-1,3- diazaspiro[4.4]non-1 -ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-( ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is administered at a dose of 200 mg-800 mg per day.

[0032] In certain embodiments of the application, the starting dose of the 4'-((2-butyl-4-oxo- 1,3-diazaspiro[4.4]non-1 -en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)- [1,1 '-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is 400 mg / day, which is increased to 800 mg / day after 14 days of continuous administration.

[0033] In preferred embodiments of the application, the compound of formula (I) or a pharmaceutically acceptable salt thereof is for oral or injectable administration, preferably in a continuous administration mode.

[0034] In preferred embodiments of the application, the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1 - en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1 '-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof is administered alone or in combination with other drugs.

[0035] In preferred embodiments of the application, the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1 - en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1 '-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof is administered alone, i.e. without the use of other drugs for the treatment or prevention of renal diseases, although the use of ancillary drugs that do not have a therapeutic or preventive effect on renal diseases or conditions is not excluded.

[0036] In preferred embodiments of the application, the renal disease does not include diabetic nephropathy or C3 glomerulopathy.

[0037] In preferred embodiments of the application, the patient with renal disease has a 24h urinary protein quantification > 0.75 g / day.

[0038] In preferred embodiments of the application, the patient with renal disease does not include proteinuria > 3.5 g / day and serum albumin < 3.0 g / dL.

[0039] In preferred embodiments of the application, the patient with renal disease has an eGFR > 30 mL / min / 1.73 m 2 .

[0040] 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5- methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof has good safety and effectiveness in the treatment of focal segmental glomerulosclerosis or immunoglobulin A nephropathy, and has few side effects.

[0041] DETAILED DESCRIPTION

[0042] Unless otherwise defined, the terms used in the present application have the following meanings:

[0043] The term "treatment" refers to administration of a compound or pharmaceutical composition described herein for the following purposes: (i) delaying the onset of a disease, i.e., causing the clinical symptoms of the disease not to appear or delaying their appearance; (ii) inhibiting the disease, i.e., preventing the development of clinical symptoms; and / or (iii) relieving the disease, i.e., causing the regression of clinical symptoms or their severity.

[0044] In the present specification, when describing a numerical range, the expressions "to", "to", "within the range" or "between the range" are used to include the end point values.

[0045] "AE" refers to adverse events.

[0046] "SAE" refers to serious adverse events.

[0047] "PK" refers to pharmacokinetics.

[0048] "PD" refers to pharmacodynamics.

[0049] "QD" refers to once a day.

[0050] "BID" refers to twice a day.

[0051] "eGFR" refers to estimated glomerular filtration rate.

[0052] "CTCAE" refers to Common Terminology Criteria for Adverse Events. DETAILED DESCRIPTION

[0053] The embodiments of the present application will be described in detail below with specific examples. The following examples are only used to illustrate the present application, and should not be regarded as limiting the scope of the present application.

[0054] Example 1 SD rat IgA nephropathy model experiment

[0055] 1.1 Construction of SD rat IgA nephropathy model

[0056] 5-6 weeks old male SD rats were divided into normal control group and modeling group according to body weight, in which 8 normal rats and 92 modeling rats. The modeling rats were given BSA intragastrically at 400 mg / kg every other day until the 13th week; subcutaneous injection of CCl4 0.1 mL + castor oil 0.4 mL once a week until the 13th week; SEB was injected into the tail vein at 0.3 mg / kg at the 11th, 12th and 13th weeks. On the 2nd day of the experiment, complete Freund's adjuvant 0.2 ml (containing BSA 2 mg) was injected subcutaneously, and on the 15th and 29th days, incomplete Freund's reagent 0.2 ml (containing BSA 2 mg) was injected intraperitoneally. The normal control group was given corresponding body weight dose of distilled water intragastrically every other day until the 13th week; subcutaneous injection of normal saline 0.5 mL once a week until the 13th week; equal amount of normal saline was injected into the tail vein at the 11th, 12th and 13th weeks. Note: start dosing during modeling (11th week).

[0057] 1.2 Test grouping

[0058] 8 normal control rats, 10 solvent control rats, 10 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound 30 mg / kg group (low dose group), 10 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound 60 mg / kg group (medium dose group), 10 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound 90 mg / kg group (high dose group).

[0059] 1.3 Drug preparation

[0060] The corresponding amount of 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-ene-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound was weighed, and the solvent (20% HP-β-CD dissolved in 0.5% (w / v) HPMC K4M) was added, mixed thoroughly, and ultrasonicated to obtain a uniform suspension.

[0061] 1.4 Administration method

[0062] Oral administration, once a day, for 8 weeks.

[0063] Normal control group and solvent control group: solvent was given;

[0064] Other administration groups: different doses of test drugs were given respectively.

[0065] 1.5 Kidney histopathology detection

[0066] (1) Kidney HE staining examination: after the last administration, kidney tissue was dissected and fixed with 10% formalin for routine HE staining histopathology examination.

[0067] (2) Kidney PAS staining examination: kidney tissue was fixed with 10% neutral formalin for PAS staining histopathology examination.

[0068] (3) Kidney glomerular IgA deposition intensity examination: kidney tissue was OCT embedded, and frozen section was used for tissue immunofluorescence staining examination to determine the expression position and strength of the target gene IgA.

[0069] 1.6 Urine total protein (UTP) and urine microalbumin (MALB) detection

[0070] Urine was collected, and the contents of urine total protein (UTP) and urine microalbumin (MALB) were detected by kit.

[0071] 1.7 Statistical analysis

[0072] The data obtained in the experiment were statistically analyzed by EXCEL and IBM SPSS Statistics 22.0.

[0073] 1.8 Experimental results

[0074] (1) Kidney HE staining: compared with the solvent control group (the kidney tissue damage score of rats was 1.90±0.18), the kidney tissue damage scores of SD rats in the low, medium and high dose groups of the drug were significantly improved, which were 1.10±0.18, 0.60±0.16 (p<0.01) and 0.60±0.22 (p<0.01) respectively.

[0075] (2) Kidney PAS staining: compared with the solvent control group (the percentage of positive expression area of kidney PAS staining was 8.81±0.70%), the percentage of positive expression area of kidney PAS staining of SD rats in the low, medium and high dose groups of the drug was significantly reduced, which were 5.72±0.34%, 5.86±0.26% and 5.81±0.31% respectively, and all had statistical significance (p<0.05).

[0076] (3) Kidney glomerular IgA deposition intensity test: compared with the solvent control group of rats, the IgA protein content in the kidney tissue of the SD rats in the low, medium and high dose groups of the drug was reduced, and the IgA fluorescence intensity was 19.17±0.47, 19.51±0.32 and 19.52±0.50, respectively.

[0077] (4) IgA nephropathy rat urine total protein (UTP) detection:

[0078] After 4 weeks of administration, the urine total protein content of the solvent control group of rats was 14.90±1.61 mg / 24h (p<0.05); the urine total protein content of the rats in the low, medium and high dose groups of the drug was reduced, and was 13.88±1.44, 13.61±1.45 and 12.74±1.16 mg / 24h, respectively.

[0079] (5) IgA nephropathy rat urine microalbumin (MALB) detection:

[0080] After 4 weeks of administration, the urine microalbumin content of the solvent control group of rats was 322.08±31.49 μg / 24h; the urine microalbumin content of the rats in the low, medium and high dose groups of the drug was 310.06±30.07, 304.87±36.13 and 269.99±26.90 μg / 24h, respectively, which were lower than that of the solvent control group.

[0081] From the above results, it can be seen that the drug used in the application can effectively improve the IgA nephropathy of SD rats.

[0082] Example 2 Multi-center clinical trial research

[0083] 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof in patients with primary immunoglobulin A nephropathy Randomized, open, positive control, multi-center clinical trial of effectiveness and safety.

[0084] 1. Research purposes

[0085] 1.1. Main research purposes

[0086] Evaluate the effectiveness and safety of 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof in patients with primary immunoglobulin A nephropathy.

[0087] 1.2. Secondary objectives

[0088] (1) To evaluate the safety and tolerability of 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1- en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof after oral administration in patients with primary IgAN.

[0089] (2) To evaluate the pharmacokinetic (PK) characteristics of 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1- en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2- sulfonamide compound or a pharmaceutically acceptable salt thereof after oral administration in patients with primary IgAN.

[0090] 2. Study population

[0091] 2.1. Patients with primary IgAN aged 18 to 65 years (inclusive).

[0092] 2.2. Subjects who can communicate well with the investigator, understand and comply with the requirements of the study, have a full understanding of the trial content, process and possible adverse reactions, and voluntarily sign the informed consent form.

[0093] 2.3. Primary IgAN confirmed by kidney biopsy.

[0094] 2.4. Stable use of RAS inhibitors at the maximum tolerated dose (at least half of the maximum permitted dose) or the maximum permitted dose for at least 12 weeks before screening (acceptable to change medication or interrupt treatment for no more than 7 consecutive days due to illness), and stable treatment until the day before the first administration of the test drug.

[0095] 2.5. Proteinuria was detected in two 24-hour urine samples during the screening period, with an interval of at least 1 week between the two tests, and the average of the two test results met: 24h-UPE ≥ 0.75 g / day.

[0096] 2.6. eGFR (based on CKD-EPI 2009 formula) ≥ 30 mL / min / 1.73m 2 .

[0097] 2.7. Screening and randomization office sitting systolic blood pressure in the range of 100 mmHg ~ 150 mmHg (including the critical value), and diastolic blood pressure in the range of 60 mmHg ~ 100 mmHg (including the critical value).

[0098] 2.8. The subjects are not in the following time period of pregnancy or lactation, no birth plan, no donation of sperm or egg plan, and voluntarily take effective contraceptive measures required by the program:

[0099] 1) Women of childbearing potential (WOCBP) from the screening period to 40 days after the last dose of the test drug;

[0100] 2) Male subjects with WOCBP partners from the screening period to 100 days after the last dose of the test drug.

[0101] 3. Test scheme

[0102] A total of 3 dose groups are pre-set, with about 30 cases in each group, and a total of about 90 subjects. The dose of 4'-((2-butyl-4-oxo-1, 3-diazaspiro [4.4] non-1-ene-3-yl) methyl-d2)-N-(4-chloro-5-methylisoxazole-3-yl)-2'-(ethoxymethyl)-[1, 1'-biphenyl]-2-sulfonamide hydrochloride compound (hereinafter referred to as formula I compound) is pre-set as 400 mg (QD), 400 mg (BID), and 300 mg (QD) of positive control drug irbesartan tablets.

[0103] The study process includes a screening period (2 weeks), a treatment observation period (12 weeks), and a follow-up period (4 weeks) / extension period (16 weeks). All subjects enrolled in the study received a maximum tolerated dose (at least half of the maximum allowable dose) or a maximum allowable dose of a renin-angiotensin system (RAS) inhibitor for at least 12 weeks before screening, but the 24h-UPE was still ≥0.75 g / day (the number of subjects with 24h-UPE ≤1 g / day was not more than 20% of the total number) at the time of screening. All subjects were stable in the use of RAS inhibitors until the day before the first dose of the test drug, and the RAS inhibitors were stopped from the first dose day (D1).

[0104] In the study, 4'-((2-butyl-4-oxo-1, 3-diazaspiro [4.4] non-1-ene-3-yl) methyl-d2)-N-(4-chloro-5-methylisoxazole-3-yl)-2'-(ethoxymethyl)-[1, 1'-biphenyl]-2-sulfonamide hydrochloride compound and irbesartan were given by titration, specifically:

[0105] All subjects will receive initial dose of trial drug (Compound of Formula I and Irbesartan) and will be dosed for 2 weeks. If tolerated (evaluated by the investigator, refer to the following criteria: 1. sitting systolic blood pressure≥90 mmHg, sitting diastolic blood pressure≥60 mmHg; 2. no investigator-considered water sodium retention, orthostatic hypotension-related symptoms or signs that are considered intolerable; 3. no investigator-considered abnormal laboratory indicators such as serum potassium >5.5 mmol / L, eGFR reduction >30% from baseline, etc.), subjects will be titrated to target dose. If not tolerated, the investigator will evaluate whether to maintain the initial dose or suspend the administration.

[0106] The initial dose of Compound of Formula I 400 mg QD, Compound of Formula I 400 mg BID and Irbesartan 300 mg QD is 200 mg QD, 200 mg BID and 150 mg QD, respectively. All subjects will be dosed continuously for 12 weeks. If a subject experiences intolerance during the study, the investigator will determine whether to reduce the dose of trial drug to the initial dose or suspend the administration, and then evaluate whether to restart the administration.

[0107] The subjects in Compound of Formula I group and the subjects in Irbesartan group who enter the extension phase will then complete a 4-week follow-up. The subjects in Irbesartan group who complete the 12-week treatment observation period can enter the extension phase to receive Compound of Formula I treatment, starting from an initial dose of 200 mg BID, and then titrating to 400 mg BID if tolerated after 2 weeks. The extension phase treatment cycle is up to 12 weeks, and then a 4-week follow-up is completed. During the study, subjects will receive safety, efficacy evaluation and PK biological sample collection at specific times according to the protocol.

[0108] 3.2. Initial dose

[0109] Compound of Formula I 200 mg QD, Compound of Formula I 200 mg BID and Irbesartan 150 mg QD.

[0110] 3.3. Duration

[0111] Compound of Formula I is administered once or twice a day for 12 consecutive weeks; Irbesartan is administered once a day for 12 consecutive weeks

[0112] 4. Dosage and administration of drugs

[0113] Compound of Formula I 200 mg QD, Compound of Formula I 200 mg BID and Irbesartan 150 mg QD, orally, after 14 days of continuous administration, the dose is adjusted to Compound of Formula I 400 mg QD, 200 mg QD, 400 mg BID or 200 mg BID.

[0114] 5. Test results

[0115] Effectiveness: one subject in the 400 mg BID group showed a 70.6% decrease in urine protein / creatinine ratio (UPCR) from baseline after 84 days of administration (i.e., day 85), and one subject in the 400 mg QD group showed a 52.1% decrease in UPCR from baseline on day 85.

[0116] Safety: the safety and tolerability of the compound of Formula I in each dose group (400 mg (QD), 400 mg (BID)) were good, most of the AEs were CTCAE grades 1-2, common AEs included decreased blood pressure, positive fecal occult blood, and decreased hemoglobin, etc., and no significant difference was found in the maximum decrease in average blood pressure after continuous administration, and no SAEs were observed. The above results suggest that the overall safety and tolerability of the drug in patients with kidney disease are good.

[0117] PK, PD characteristics: the compound of Formula I has strong inhibitory activity on endothelin 1 and angiotensin II related signal pathways, and good effect on improving proteinuria and delaying eGFR progression.

[0118] The results of the study show that the compound of Formula I can effectively improve the urine protein of IgAN patients, and has excellent efficacy and safety.

Claims

1. Use of a 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment or prevention of a renal disease, preferably a renal disease selected from the group consisting of focal segmental glomerulosclerosis or immunoglobulin A nephropathy.

2. Use according to claim 1, characterized in that, 4. The pharmaceutically acceptable salt of the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound is selected from the group consisting of hydrochloride, sulfate, nitrate, hydrobromide, hydrofluoride, hydroiodide, phosphate, 2,5-dihydroxybenzoate, 1-hydroxy-2-naphthoate, acetate, dichloroacetate, trichloroacetate, acetyloxymethane, adipate, benzenesulfonate, 4-chlorobenzenesulfonate, benzoate, 4-acetamidobenzoate, 4-aminobenzoate, decanoate, hexanoate, octanoate, cinnamoate, citrate, cyclohexanesulfamate, camphorsulfonate, aspartate, camphorate, gluconate, glucaronate, glutamate, isoascorbate, lactate, malate, mandelate, pyroglutamate, tartrate, dodecylsulfate, dibenzoyl tartrate, ethane-1,2-disulfonate, ethanesulfonate, formate, fumarate, galacturonate, gentisate, glutarate, 2-ketoglutarate, glycolate, hippurate, isethionate, lactobionate, ascorbate, aspartate, laurate, camphorate, maleate, malonate, mesylate, 1,5-naphthalene disulfonate, naphthalene-2-sulfonate, nicotinate, oleate, orotate, oxalate, palmitate, pamoate, propionate, salicylate, 4-aminosalicylate, sebacate, stearate, succinate, thiocyanate, pamoate, formate, undecylenate, trifluoroacetate, benzenesulfonate, p-toluenesulfonate or L-malate; preferably mesylate, benzenesulfonate, isethionate, ethanesulfonate, hydrochloride, sulfate, p-toluenesulfonate, phosphate or hydrobromide, more preferably hydrochloride.

3. Use according to claim 1, characterized in that, The single dose of the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is in the range of 1-1500 mg, preferably 10-1200 mg, 20-1100 mg, 30-1000 mg, 40-900 mg, 50-800 mg, 60-750 mg, 70-700 mg, 80-650 mg, 90-600 mg, 100-550 mg, 150-500 mg, 200-450 mg, 250-400 mg, 200-400 mg, or 300-350 mg; more preferably 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 95 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg; further preferably 100 mg, 200 mg, or 400 mg.

4. Use according to claim 1, characterized in that, The 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is for oral administration or injection, preferably in a continuous administration manner.

5. Use according to claim 1, characterized in that, The 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is administered once a day, twice a day, or three times a day.

6. Use according to claim 1, characterized in that, The single dose of the 4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl-d2)-N-(4-chloro-5-methylisoxazol-3-yl)-2'-(ethoxymethyl)-[1,1'-biphenyl]-2-sulfonamide compound or a pharmaceutically acceptable salt thereof is 200 mg or 400 mg, once a day or twice a day; preferably, the single dose is 200 mg, once or twice a day, and the dose is increased to 400 mg, once or twice a day, after 14 days of continuous administration; more preferably, the single dose is 200 mg, once a day, and the dose is increased to 400 mg, once a day, after 14 days of continuous administration, or the single dose is 200 mg, twice a day, and the dose is increased to 400 mg, twice a day, after 14 days of continuous administration.

7. Use according to claim 1, characterized in that, The kidney disease does not include diabetic nephropathy or C3 glomerulopathy.

8. Use according to claim 1, characterized in that, The patient with kidney disease has a 24h urine protein quantification of ≥0.35 g / day, or ≥0.5 g / day, or even ≥0.75 g / day.

9. Use according to claim 1, characterized in that, Patients with renal disease do not include those with proteinuria > 3.5 g / day and serum albumin < 3.0 g / dL.

10. Use according to claim 1, characterized in that, Patients with renal disease eGFR > 30 mL / min / 1.73 m 2 .

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