Application of JAK2 inhibitor in preparation of medicine for treating chronic kidney disease

Abstract

The invention relates to an application of a JAK inhibitor in preparation of a medicine for treating chronic kidney diseases, in particular to a compound 2-{N-[4-(4-methyl) piperazine] phenyl} amino-5, 6-dihydro-7-[3-(N-tert-butyl) sulfonamido] phenyl-pyrrolo [2, 3-d] pyrimidine or pharmaceutically acceptable salt thereof, and a preparation method of the compound 2-{N-[4-(4-methyl) piperazine] phenyl} amino-5, 6-dihydro-7-[3-(N-tertiary butyl) sulfonamido] phenyl-pyrrolo [2, 3-d] pyrimidine. The invention also relates to application of a pharmaceutical composition containing the compound in preparation of medicines for treating chronic kidney diseases. The compound has good treatment activity on chronic kidney diseases caused by various reasons, can remarkably reduce the inflammatory factor level of kidney tissue, further reduces kidney injury and fibrosis lesion degree, is high in safety, is expected to become a new medicine for treating the chronic kidney diseases, and has wide clinical application prospects.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and relates to use of a JAK2 inhibitor in preparing a drug for treating chronic kidney disease, and specifically relates to a compound 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine or a pharmaceutically acceptable salt thereof, and use of a pharmaceutical composition containing the same in preparing a drug for treating chronic kidney disease. Background Art

[0002] Chronic kidney disease (CKD) is a disease characterized by abnormal kidney structure or function for more than 3 months due to various reasons, including signs of kidney damage (albuminuria, abnormal urine sediment, tubular-related lesions, abnormal histological examinations, and abnormal imaging examinations) or a history of kidney transplantation, with or without a decrease in glomerular filtration rate (GFR); or an unexplained decrease in GFR (<60 ml / min) for more than 3 months. It is estimated that there are 850 million kidney disease patients worldwide, and the prevalence of CKD in my country is 9.4% to 12.1%, with the number of patients reaching 120 million. CKD accounts for a considerable proportion of the global disease burden and is still growing; approximately 10% of adults worldwide are affected by some form of chronic kidney disease, resulting in 1.2 million deaths each year. With the increasing proportion of diabetes, hypertension, and metabolic syndrome in the elderly population, the incidence of kidney disease will further deteriorate, and CKD is expected to become the fifth leading cause of death in the world by 2040.

[0003] Diabetic nephropathy (DKD) can be considered as a type of CKD. It is a chronic kidney disease associated with diabetes. It is a clinical syndrome characterized by persistent albuminuria and a decrease in estimated glomerular filtration rate (eGFR). It is an important cause of end-stage renal disease in my country and most developed countries, and is associated with a significant increase in the incidence and mortality of cardiovascular diseases. A survey conducted by the Chinese Diabetes Society in 2020 showed that 20% to 40% of diabetic patients in my country have diabetic nephropathy. If not treated in time, kidney disease will be irreversible, and drug treatment is ineffective, resulting in a poor prognosis.

[0004] JAK / STATs is the abbreviation of tyrosine protein kinase (JAK) / signal transducer and activator of transcription (STAT) pathway. It is widely involved in various physiological and pathological processes of cells, mediating cell reproduction differentiation, immune regulation, migration and apoptosis, etc. It is an important signaling pathway for maintaining body homeostasis and is closely related to multiple organs. As far as the kidney is concerned, JAK / STATs plays an important role in primary kidney disease, secondary kidney damage, and complications of chronic kidney disease.

[0005] At present, the research targeting JAK / STATs mainly focuses on the pathological processes such as glomerular fibrosis, renal interstitial fibrosis, tubular epithelial cell transdifferentiation, lipid metabolism and absorption, and inflammation mediated by JAK / STATs, and the application of clinical targeted therapy is relatively rare. Although there are first-line therapies such as angiotensin-converting enzyme inhibitors (ACEI), type I angiotensin II receptor blockers (ARB), and sodium-glucose cotransporter 2 inhibitors (SGLT2) for the treatment of chronic kidney disease and diabetic nephropathy, the research and development of innovative drugs with new mechanisms and new targets still has great social significance and market prospects.

[0006] Future drug development should slow disease progression at an early stage and prevent the development of end-stage renal failure (ESRD) and cardiovascular complications. Persistent low-grade inflammation is considered an important component of chronic kidney disease and plays a unique role in its pathophysiology. Evidence of inflammatory pathway activation can be found both in the early and late stages of the disease. Therefore, JAK-STATs inhibitors that inhibit important inflammatory signaling pathways, such as JAK1 inhibitors and JAK2 inhibitors, are expected to play an important role in delaying the progression of chronic kidney disease and diabetic nephropathy.

[0007] A phase II clinical trial studied the effect of barcitinib, a dual-target inhibitor of JAK1 and JAK2, on diabetic nephropathy. The results showed that barcitinib can significantly reduce proteinuria in patients with diabetic nephropathy and has a good clinical therapeutic effect on diabetic nephropathy.

[0008] WO2014085154A1 discloses a method for treating kidney disease. Specifically disclosed is a method for treating or reducing the likelihood of developing kidney disease, comprising administering to a subject in need thereof an agent that reduces the expression level of a pathogenic APOL1 polypeptide encoded by an APOL1 risk allele relative to the APOL1 polypeptide expression level in the absence of the agent. The agent is an antagonist of the Janus kinase (JAK) / signal transducer and activator of transcription (STAT) pathway. The (JAK / STAT pathway) antagonist targets at least one JAK selected from JAK1, JAK2, and JAK3. The agent is selected from INCB-018424, TG101348, WHI-P131, LY3009104, INCB-28050, SB1518, lecithinib, ruxolitinib, CYT387, LY3009104, SD1008, cucurbitacin, GO6976, WHI-P154, AG490 and CP-690550.

[0009] In September 2021, the U.S. Food and Drug Administration (FDA) required all approved JAK inhibitor drugs to add a black box warning in the instructions for increased risk of serious heart-related events, cancer, thrombosis, and death. By improving the selectivity of JAK2, higher therapeutic activity can be achieved while avoiding the side effects of JAK1 inhibitor drugs, such as severe cytopenia, infection, and secondary second tumors. Summary of the invention

[0010] In view of the lack of highly selective JAK2 inhibitors that can inhibit important inflammatory signaling pathways in existing clinical treatment technologies for chronic kidney disease, the present invention aims to make up for the shortcomings of existing drug treatments and provide new therapeutic drugs with more new mechanisms and new targets for clinical treatment. The present invention provides a use of a JAK2 inhibitor in the preparation of a drug for the treatment of chronic kidney disease, and specifically relates to the use of the compound 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing the same in the preparation of a drug for the treatment of chronic kidney disease.

[0011] The inventors found in previous studies that the compound shown in Formula 1 showed a highly selective JAK2 inhibitory effect. Based on this research and the above background, the inventors conducted in-depth research on the treatment of chronic kidney disease with this type of compound. Experimental studies have confirmed that this type of compound has good therapeutic activity for chronic kidney disease caused by various reasons, can significantly reduce the level of inflammatory factors in kidney tissue, thereby reducing kidney damage and fibrosis, and has high safety. It is expected to become a new drug for the treatment of chronic kidney disease and has broad clinical application prospects.

[0012] The present invention is achieved through the following technical solutions:

[0013] The first aspect of the present invention provides the use of a compound of formula 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating chronic kidney disease;

[0014]

[0015] Optionally, the chronic kidney disease refers to a syndrome consisting of a decrease in glomerular filtration rate caused by various chronic kidney diseases and the metabolic disorders and clinical symptoms associated therewith, wherein the renal function damage is progressive and irreversible, eventually leading to complete loss of renal function; its specific diseases or conditions include primary chronic kidney disease, secondary chronic kidney disease, chronic tubulointerstitial lesions, or hereditary kidney disease.

[0016] Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, or chronic nephritis that develops after prolonged acute or rapidly progressive nephritis; the secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy, or lupus erythematosus nephropathy; the diabetic nephropathy includes chronic kidney disease associated with patients with type 1 and type 2 diabetes; the chronic tubulointerstitial lesions include chronic pyelonephritis or chronic uric acid nephropathy.

[0017] Optionally, the pharmaceutically acceptable salt is an inorganic salt or an organic salt; the inorganic salt or organic salt may be conventionally used in the art. Optionally, the inorganic salt preferably includes hydrochloride, sulfate, phosphate, or nitrate; the organic salt preferably includes methanesulfonate, citrate, oxalate, or succinate.

[0018] The second aspect of the present invention provides a use of a pharmaceutical composition in the preparation of a drug for treating chronic kidney disease, characterized in that the pharmaceutical composition comprises a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier;

[0019]

[0020] Optionally, the compound of formula 1 or a pharmaceutically acceptable salt thereof in the pharmaceutical composition accounts for 0.01 to 90% of the mass of the pharmaceutical composition; preferably, the compound of formula 1 or a pharmaceutically acceptable salt thereof in the pharmaceutical composition accounts for 0.05 to 70% of the mass of the pharmaceutical composition.

[0021] Optionally, the pharmaceutically acceptable salt is an inorganic salt or an organic salt; the inorganic salt or organic salt may be conventionally used in the art. Optionally, the inorganic salt preferably includes hydrochloride, sulfate, phosphate, or nitrate; the organic salt preferably includes methanesulfonate, citrate, oxalate, or succinate.

[0022] Optionally, the pharmaceutically acceptable carrier is selected from one or more of fillers, glidants, lubricants, binders, disintegrants, and solvents; optionally, the filler is selected from one or more of mannitol, microcrystalline cellulose, starch, sucrose, dextrin, lactose, powdered sugar, and glucose; the glidant is selected from one or more of talc and colloidal silicon dioxide; the lubricant is selected from one or more of magnesium stearate, sodium stearyl fumarate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, and poloxamer; the binder is selected from one or more of water, ethanol, starch slurry, syrup, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, and polyvinyl pyrrolidone; the disintegrant is selected from one or more of cross-linked polyvinylpyrrolidone, starch, sodium bicarbonate, citric acid, tartaric acid, and low-substituted hydroxypropyl cellulose; the solvent includes water and / or a balanced salt solution.

[0023] Optionally, the pharmaceutically acceptable carrier further comprises a flavoring agent or a sweetener.

[0024] Optionally, the pharmaceutical composition comprising the compound of formula 1 and a pharmaceutically acceptable salt thereof is an oral preparation.

[0025] Optionally, the oral preparation includes tablets, capsules, granules or oral solutions.

[0026] On the other hand, the present invention also provides a method for preventing or treating chronic kidney disease, comprising administering to a subject (e.g., a human or an animal) in need thereof an effective amount of the compound of formula 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition. Optionally, the chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic tubulointerstitial lesions, or hereditary nephropathy.

[0027] Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, or chronic nephritis that develops from prolonged acute or rapidly progressive nephritis; the secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy, or lupus erythematosus nephropathy; the diabetic nephropathy includes chronic kidney disease associated with patients with type 1 and type 2 diabetes; the chronic tubulointerstitial lesions include chronic pyelonephritis or chronic uric acid nephropathy.

[0028] The expression "effective amount" or "therapeutically effective amount" in the present invention includes an amount sufficient to alleviate or prevent the symptoms or conditions of a medical disease. "Effective amount" also refers to an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject can vary depending on factors such as the disease to be treated, the patient's general health, the method, route and dosage of administration, and the severity of side effects. The effective amount can be a maximum dose or a dose that avoids significant side effects or toxic effects.

[0029] Optionally, the therapeutically effective amount of the compound of formula 1 or a pharmaceutically acceptable salt thereof administered is 10 mg to 1000 mg; optionally, the therapeutically effective amount of the pharmaceutical composition administered is 10 mg to 1000 mg.

[0030] On the other hand, the present invention also provides a highly selective JAK2 inhibitor for preventing or treating chronic kidney disease, wherein the chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic tubulointerstitial disease, or hereditary kidney disease.

[0031] Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, or chronic nephritis that develops from prolonged acute or rapidly progressive nephritis; the secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy, or lupus erythematosus nephropathy; the diabetic nephropathy includes chronic kidney disease associated with patients with type 1 and type 2 diabetes; the chronic tubulointerstitial lesions include chronic pyelonephritis or chronic uric acid nephropathy.

[0032] The present invention has the following outstanding effects:

[0033] (1) The compound of formula 1 or its pharmaceutically acceptable salt is used as the sole active ingredient of the drug, which can significantly reduce the level of inflammatory factors in kidney tissue, thereby reducing the degree of kidney fibrosis, and is used to treat chronic kidney disease.

[0034] (2) Compared with the current first-line clinical drugs for the treatment of chronic kidney disease (irbesartan and canagliflozin), the compound of formula 1 has a more significant clinical advantage in terms of efficacy, mainly manifested in a lower onset dose and stronger therapeutic activity at the same dose (see Examples 1 to 4 for details).

[0035] (3) The compound of formula 1 has a more significant advantage in pharmacological activity in treating chronic kidney disease than the JAK2 selective inhibitor ferritinib that has been marketed.

[0036] (4) The compound of formula 1 has a higher selectivity for the JAK2 target than baricitinib, a dual-target inhibitor of JAK1 and JAK2 that has been tested in phase II clinical studies for diabetic nephropathy. The inhibitory activity of the compound of formula 1 against JAK2 is 99 times that of the JAK1 subtype (see patent CN110305140A for details), and the literature reports (“Pharmacological mechanism and clinical application study of Janus kinase inhibitors baricitinib and upadacitinib”, Chinese Journal of Clinical Pharmacology, 2023, 39(18): 2713-2717) that the IC values ​​of baricitinib for JAK1 and JAK2 inhibition are 50 The selectivity of JAK2 is 5.9nM and 5.7nM respectively, with no subtype selectivity. By improving the selectivity of JAK2, higher therapeutic activity can be obtained, while avoiding the side effects of JAK1 inhibitor drugs, such as severe cytopenia, infection, and secondary second tumors, with the advantages of better activity and higher safety. DETAILED DESCRIPTION

[0037] The present invention is further described in detail below in conjunction with specific implementation modes.

[0038] Example 1: Study on the pharmacodynamic activity of the compound of formula 1 on the unilateral ureteral occlusion (UUO) model in rats

[0039] The unilateral ureteral ligation-induced renal interstitial fibrosis model in rats was used to investigate the therapeutic effect of the compound of formula 1 on renal interstitial fibrosis.

[0040] 70 SD rats were randomly divided into 7 groups according to body weight, namely, sham operation group, model control group, irbesartan group (25.0 mg / kg), fedratinib group (6.0 mg / kg), low-dose, medium-dose and high-dose groups (2.5, 5.0 and 10.0 mg / kg) of the compound of formula 1, with 10 animals in each group. The renal interstitial fibrosis model was established by ligation of the ureter near the left renal pelvis; except that the ligation was not performed on the rats in the sham operation group, the rest of the operations were the same as those of the rats in the model control group. The drug was administered on the day of modeling, once a day, for 14 consecutive days. On the next day after the last administration, blood was collected from the abdominal aorta of the rats in each group after anesthesia to detect renal function indicators (blood CRE and BUN); the left kidney tissue was dissected and the contents of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and transforming growth factor β1 (TGF-β1) in the tissue were detected by ELISA kits; the left kidney was dissected and HE staining and Masson staining were performed respectively. HE staining was used to observe the degree of renal tissue damage and score it, and Masson staining was used to observe the degree of renal tissue fibrosis and score it.

[0041] Experimental results:

[0042] 1. Effect on renal function: As can be seen from Table 1 below, the medium and high doses (5.0 and 10.0 mg / kg) of the compound of formula 1 can significantly reduce the CRE level and BUN level in the blood of model rats. At the same dose (10.0 mg / kg), the pharmacodynamic activity of the compound of formula 1 is significantly better than that of the positive control drug (irbesartan). The CRE level and BUN level in the blood of rats in the fedratinib group showed a downward trend, but there was no statistical difference compared with the model control group. Therefore, the activity of the compound of formula 1 in treating chronic kidney disease is significantly better than that of irbesartan and fedratinib.

[0043] Table 1: Effects of the compound of formula 1 on renal function indices in SD rats with UUO model ( n=10)

[0044]

[0045] Note: Compared with sham operation group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.

[0046] 2. Effect on renal tissue cytokines: As can be seen from Table 2 below, the medium and high doses (5.0, 10.0 mg / kg) of the compound of formula 1 can significantly reduce the levels of IL-6 and TNF-α in the renal tissue of model rats, and the low, medium and high doses (2.5, 5.0, 10.0 mg / kg) of the compound of formula 1 can significantly reduce the level of TGF-β1 in the renal tissue of model rats. At the same dose (10.0 mg / kg), the pharmacodynamic activity of the compound of formula 1 is better than that of the positive control drug (irbesartan). The levels of IL-6 and TNF-α in the fedratinib group showed a downward trend, but there was no statistical difference compared with the model control group. Therefore, the activity of the compound of formula 1 in treating chronic kidney disease is significantly better than that of irbesartan and fedratinib.

[0047] Table 2: Effects of the compound of formula 1 on IL-6, TNF-α, and TGF-β1 in UUO model of SD rats ( n=10)

[0048]

[0049]

[0050] Note: Compared with sham operation group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.

[0051] 3. Effect on morphological changes of renal tissue: As can be seen from Tables 3 and 4 below, low, medium and high doses (2.5, 5.0, 10.0 mg / kg) of the compound of formula 1 can improve the tubular dilatation, renal interstitial inflammation, tubular atrophy and renal cortical tubular and interstitial fibrosis of model rats, among which, medium and high doses (5.0, 10.0 mg / kg) can significantly reduce the renal tissue lesion scores (HE staining and Masson staining) of model rats. At the same dose (10.0 mg / kg), the pharmacodynamic activity of the compound of formula 1 is better than that of the positive control drug (irbesartan). The fedratinib group has a certain improvement, but there is no statistical difference compared with the model control group. Therefore, the activity of the compound of formula 1 in treating chronic kidney disease is significantly better than that of irbesartan and fedratinib.

[0052] Table 3: Effects of the compound of formula 1 on the HE staining pathological scores of UUO model in SD rats ( n=10)

[0053]

[0054] Note: Compared with sham operation group ++ P≤0.01; compared with the model control group ** P≤0.01,* P≤0.05.

[0055] Table 4: Effects of the compound of formula 1 on Masson staining pathology in UUO model of SD rats ( n=10)

[0056]

[0057]

[0058] Note: Compared with sham operation group ++ P≤0.01; compared with the model control group ** P≤0.01.

[0059] The above research results show that the compound of formula 1 has a significant therapeutic effect on the UUO model of rats, can reduce the level of inflammatory factors in renal tissue, and thus reduce the degree of renal fibrosis. At the same dose, the activity of the compound of formula 1 in treating chronic kidney disease is significantly better than that of irbesartan and fedratinib.

[0060] Example 2: Study on the pharmacodynamic activity of the compound of formula 1 on the rat chronic kidney disease model induced by adriamycin

[0061] The rat chronic kidney disease model induced by doxorubicin was used to investigate the therapeutic effect of the compound of formula 1 on chronic kidney disease. Since fedratinib failed to show therapeutic activity on chronic kidney disease in the rat unilateral ureteral ligation (UUO) model (multiple indicators showed no statistical difference compared with the model control group), fedratinib was no longer used as a positive control drug in this model, and only the clinical first-line treatment drug irbesartan was used as a positive control drug.

[0062] SD rats were injected with 3 mg / kg doxorubicin via the tail vein once a week for 4 consecutive weeks, and observed for 4 weeks to induce a rat chronic kidney disease model. The rats with successful models were selected and randomly divided into model control group, irbesartan group (25.0 mg / kg), low-dose, medium-dose and high-dose groups of the compound of formula 1 (3.0, 9.0, 27.0 mg / kg), and the drugs were administered once a day for 8 consecutive weeks.

[0063] The serum urea nitrogen (BUN), blood creatinine (CRE) levels, urine creatinine and urine protein of each group of animals were measured before administration and 2, 4, 6 and 8 weeks after administration, and the creatinine clearance rate was calculated. The right kidney tissue was dissected on the day after the last administration, and the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) in the kidney tissue were detected by rat ELISA kit. The left kidney was dissected, sliced, embedded, and stained with HE and PAS, respectively, to observe the morphological changes of kidney tissue and investigate the pathological changes of kidney tissue (degree of kidney damage and degree of fibrosis).

[0064] Experimental results:

[0065] 1. Effect on renal function: As shown in Table 5 below, the compound of formula 1 can significantly reduce the serum CRE level and BUN level of the model rats, the effective dose is a low dose (3.0 mg / kg), and the onset time is 6 weeks of administration; the compound of formula 1 can significantly increase the creatinine clearance rate of the model rats, the effective dose is a low dose (3.0 mg / kg), and the onset time is 6 weeks of administration. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drug (irbesartan).

[0066] Table 5: Effects of the compound of formula 1 on renal function in SD rats at the 6th week of administration ( n=8~10)

[0067]

[0068] Note: Compared with the normal control group ++ P≤0.01, + P≤0.05; compared with the model control group ** P≤0.01, * P≤0.05.

[0069] 2. Effect on renal tissue cytokines: As shown in Table 6 below, low, medium and high doses (3.0, 9.0 and 27.0 mg / kg) of the compound of formula 1 can significantly reduce the levels of IL-6 and TNF-α in model rats, and the high dose (27.0 mg / kg) of the compound of formula 1 can significantly reduce the level of TGF-β1 in rats. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drug (irbesartan).

[0070] Table 6: Effects of the compound of formula 1 on cytokines in kidney tissue of SD rats ( n=8~10)

[0071]

[0072] Note: Compared with the normal control group ++ P≤0.01; compared with the model control group** P≤0.01, * P≤0.05.

[0073] 3. Effect on renal tissue: As shown in Tables 7 and 8 below, low, medium and high doses (3.0, 9.0, 27.0 mg / kg) of the compound of Formula 1 can reduce the degree of renal interstitial inflammation, glomerular atrophy and tubular vacuolar degeneration in model rats, and medium and high doses (9.0, 27.0 mg / kg) can significantly reduce the degree of renal tissue fibrosis in model rats and significantly reduce the PAS staining score of the kidneys in model rats. At the same dose, the activity of the compound of Formula 1 is better than that of the positive control drug (Irbesartan).

[0074] Table 7: Effects of the compound of formula 1 on HE staining of kidney tissue in SD rats ( n=8~10)

[0075]

[0076] Note: Compared with the normal control group ++ P≤0.01, + P≤0.05; compared with the model control group * P≤0.05, ** P≤0.01.

[0077] Table 8: Effects of the compound of formula 1 on PAS staining scores of kidney tissue in SD rats ( n=8~10)

[0078]

[0079]

[0080] Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.

[0081] The above research results show that the compound of formula 1 has a significant therapeutic effect on the chronic kidney disease model induced by doxorubicin in rats. Its effective dose is a low dose (3.0 mg / kg), which can reduce the degree of kidney lesions and reduce the level of local tissue inflammatory factors. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drug (irbesartan).

[0082] Example 3: Study on the pharmacodynamic activity of the compound of formula 1 on the streptozotocin (STZ)-induced diabetic nephropathy model in rats

[0083] The STZ-induced rat type 1 diabetic nephropathy model was used to investigate the therapeutic effect of the compound of formula 1 on diabetic nephropathy.

[0084] The diabetic model of rats was induced by intraperitoneal injection of 70 mg / kg STZ; rats with successful model were selected and randomly divided into model control group, irbesartan group (25.0 mg / kg), fedratinib group (9.0 mg / kg), canagliflozin group (9.0 mg / kg), low-dose, medium-dose and high-dose groups of Formula 1 (3.0, 9.0 and 27.0 mg / kg), 10 rats / group according to 24h urinary microalbumin excretion. The drug was administered once a day for 8 consecutive weeks.

[0085] Urinary CRE levels, urine microalbumin, serum BUN and CRE levels were detected at the 4th, 6th and 8th weeks of administration, and the 24-h urine microalbumin excretion, the ratio of urine microalbumin / urine CRE, the ratio of urine CRE / blood CRE and urine creatinine clearance were calculated. The animals in each group were anesthetized and euthanized the day after the last administration, and the kidney tissues were dissected and the degree of kidney tissue damage was observed and scored by HE staining, and the degree of kidney tissue basement membrane sclerosis was observed and scored by PAS staining.

[0086] Experimental results:

[0087] 1. Effect on renal function: As shown in Table 9 below, the compound of formula 1 can significantly reduce the ratio of urine microalbumin to urine CRE, increase the ratio of urine CRE to blood CRE and creatinine clearance in the SD rat diabetic nephropathy model. The effective dose is a low dose (3.0 mg / kg) and the effective time is 6 weeks of administration. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drugs (irbesartan, canagliflozin, fedratinib).

[0088] Table 9 Effects of the compound of formula 1 on renal function of SD rats in the 6th week of administration ( n=10)

[0089]

[0090] Note: Compared with the normal control group ++ P≤0.015; compared with the model control group ** P≤0.01, * P≤0.05.

[0091] 2. Effect on renal tissue: As shown in Tables 10 and 11 below, the compound of formula 1 can significantly improve the pathological changes of renal tissue, reduce the degree of tubular degeneration and necrosis, renal interstitial inflammatory cell infiltration, and renal cortical and interstitial fibrosis, and can significantly reduce the HE staining score of renal tissue and the PAS staining score of glomeruli. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drug (irbesartan, canagliflozin). The fedratinib group has a certain improvement, but there is no statistical difference compared with the model control group.

[0092] Table 10: Effects of the compound of formula 1 on the pathological scores of HE staining in SD rat diabetic nephropathy model ( n=10)

[0093]

[0094]

[0095] Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01.

[0096] Table 11: Effects of the compound of formula 1 on PAS staining pathological scores in SD rat diabetic nephropathy model ( n=10)

[0097]

[0098] Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.

[0099] The above research results show that the compound of formula 1 has a significant therapeutic effect on the STZ-induced rat type 1 diabetic nephropathy model, and its effective dose is a low dose (3.0 mg / kg), which can significantly reduce the degree of renal tissue lesions. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drugs (irbesartan, canagliflozin, fedratinib).

[0100] Example 4: Study on the pharmacodynamic activity of the compound of formula 1 on the db / db mouse diabetic nephropathy model

[0101] The spontaneous type 2 diabetic nephropathy model of db / db mice was used to investigate the therapeutic effect of the compound of formula 1 on diabetic nephropathy. Since fedratinib failed to show therapeutic activity on diabetic nephropathy in the streptozotocin (STZ)-induced rat diabetic nephropathy model (no statistical difference in pathological score compared with the model control group), fedratinib was no longer used as a positive control drug in this model, and only the clinical first-line treatment drugs irbesartan and canagliflozin were selected as positive control drugs.

[0102] DB / DB mice were used and divided into model control group, irbesartan group (39.0 mg / kg), canagliflozin group (13.0 mg / kg), low-dose, medium-dose and high-dose groups (4.3, 13.0 and 39.0 mg / kg) of the compound of formula 1 by stratified random method according to 24h urinary microalbumin excretion taking into account blood glucose and body weight, 10 mice per group, once a day for 9 consecutive weeks.

[0103] 24h urine was collected from mice in each group at the 4th, 6th, and 9th weeks of administration to detect urine CRE and urine microalbumin concentrations. Orbital blood was collected to detect serum CRE and BUN. The 24h urine microalbumin excretion, urine microalbumin / urine CRE ratio, and urine creatinine clearance were calculated based on serum CRE, urine CRE, urine microalbumin concentration, and 24h urine volume. On the day after the last administration, the right renal cortex was obtained, and the degree of renal tissue damage was observed and scored by HE staining, and the degree of renal tissue basement membrane sclerosis was observed and scored by PAS staining.

[0104] Experimental results:

[0105] 1. Effect on renal function: As shown in Table 12 below, the compound of formula 1 can significantly reduce the 24h urine microalbumin excretion, increase the urine creatinine to blood creatinine ratio, and increase the creatinine clearance rate in the db / db mouse diabetic nephropathy model. The effective dose is the medium dose (13.0 mg / kg) and the effective time is 9 weeks of administration. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drugs (irbesartan and canagliflozin).

[0106] Table 12: Effects of the compound of formula 1 on renal function in db / db mice at 9 weeks after administration ( n=7~10)

[0107]

[0108] Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.

[0109] 2. Effect on renal tissue: As shown in Tables 13 and 14 below, low, medium and high doses (4.33, 13.0, 39.0 mg / kg) of the compound of Formula 1 can significantly reduce the HE staining score of renal tissue in the db / db mouse diabetic nephropathy model and significantly reduce the PAS staining score of glomeruli. The effective dose is the low dose (4.33 mg / kg). At the same dose, the activity of the compound of Formula 1 is better than that of the positive control drugs (irbesartan and canagliflozin).

[0110] Table 13: Effects of the compound of formula 1 on the pathological scores of HE staining of kidneys in db / db mice ( n=9~10)

[0111]

[0112] Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01,* P≤0.05.

[0113] Table 14: Effects of the compound of formula 1 on the PAS staining pathological scores of db / db mouse glomeruli ( n=9~10)

[0114]

[0115] Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.

[0116] The above research results show that the compound of formula 1 has a significant therapeutic effect on the spontaneous type 2 diabetic nephropathy model of db / db mice. Its effective dose is a low dose (4.3 mg / kg), which can significantly reduce the degree of renal tissue lesions. At the same dose, the activity of the compound of formula 1 is better than that of the positive control drugs (irbesartan and canagliflozin).

[0117] The above-mentioned embodiments only express some specific implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Use of a compound of formula 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating chronic kidney disease, 2. The use according to claim 1, characterized in that The chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic tubulointerstitial disease, or hereditary kidney disease; Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, or chronic nephritis that develops after prolonged acute or rapidly progressive nephritis; The secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy, or lupus erythematosus nephropathy; the diabetic nephropathy includes chronic kidney disease associated with patients with type 1 or type 2 diabetes; The chronic tubulointerstitial disease includes chronic pyelonephritis or chronic uric acid nephropathy.

3. The use according to claim 1 or 2, characterized in that The pharmaceutically acceptable salt is an inorganic salt or an organic salt; Optionally, the inorganic salt includes hydrochloride, sulfate, phosphate, or nitrate; and the organic salt includes methanesulfonate, citrate, oxalate, or succinate.

4. Use of a pharmaceutical composition in the preparation of a drug for treating chronic kidney disease, characterized in that: The pharmaceutical composition comprises a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

5. The use according to claim 4, characterized in that: The chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic tubulointerstitial disease, or hereditary kidney disease; Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, or chronic nephritis that develops after prolonged acute or rapidly progressive nephritis; The secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy, or lupus erythematosus nephropathy; the diabetic nephropathy includes chronic kidney disease associated with patients with type 1 or type 2 diabetes; The chronic tubulointerstitial disease includes chronic pyelonephritis or chronic uric acid nephropathy.

6. The use according to claim 4 or 5, characterized in that The pharmaceutically acceptable carrier includes one or more of a filler, a glidant, a lubricant, a binder, a disintegrant, and a solvent; Optionally, the filler is selected from one or more of mannitol, microcrystalline cellulose, starch, sucrose, dextrin, lactose, powdered sugar, and glucose; the glidant is selected from one or more of talc and colloidal silicon dioxide; the lubricant is selected from one or more of magnesium stearate, sodium stearyl fumarate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, and poloxamer; the binder is selected from one or more of water, ethanol, starch slurry, syrup, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, and polyvinyl pyrrolidone; the disintegrant is selected from one or more of cross-linked polyvinylpolypyrrolidone, starch, sodium bicarbonate, citric acid, tartaric acid, and low-substituted hydroxypropyl cellulose; the solvent includes water and / or a balanced salt solution; the pharmaceutically acceptable carrier also includes a flavoring agent or a sweetener.

7. The use according to any one of claims 4 to 6, characterized in that: The pharmaceutically acceptable salt is an inorganic salt or an organic salt; Optionally, the inorganic salt includes hydrochloride, sulfate, phosphate, or nitrate; and the organic salt includes methanesulfonate, citrate, oxalate, or succinate.

8. The use according to any one of claims 4 to 7, characterized in that: The dosage form of the pharmaceutical composition is an oral preparation.

9. The use according to claim 8, characterized in that The oral preparations include tablets, capsules, granules, or oral solutions.

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