Use of JAK2 inhibitor in preparation of drug for treating chronic kidney disease
By developing highly selective JAK2 inhibitor compounds, the problem of the lack of effective inhibitors in the treatment of chronic kidney disease has been solved, achieving significant reduction in renal fibrosis and side effects, and providing higher therapeutic activity and safety.
Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Filing Date
- 2025-09-11
- Publication Date
- 2026-04-02
AI Technical Summary
The current clinical treatment of chronic kidney disease lacks effective JAK2 highly selective inhibitors, resulting in poor treatment outcomes, and existing drugs have side effects such as severe blood cell reduction and infection risk.
To develop a highly selective JAK2 inhibitor compound, 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine and its pharmaceutically acceptable salt, for the preparation of a medicament for the treatment of chronic kidney disease, which alleviates fibrotic lesions by reducing the levels of inflammatory factors in renal tissue.
It significantly reduces the level of inflammatory factors in kidney tissue, alleviates the degree of kidney fibrosis, and has higher therapeutic activity and safety. It is superior to existing first-line clinical drugs and JAK1/JAK2 dual-target inhibitors, and avoids the side effects of JAK1 inhibitors.
Smart Images
Figure PCTCN2025120743-FTAPPB-I100001 
Figure PCTCN2025120743-FTAPPB-I100002 
Figure PCTCN2025120743-FTAPPB-I100003
Abstract
Description
Use of JAK2 inhibitor in the preparation of a medicament for treating chronic kidney disease
[0001] Priority information
[0002] The present application claims priority to and the benefit of Chinese Patent Application No. CN202411340377.1, filed September 25, 2024, and is hereby incorporated by reference in its entirety. TECHNICAL FIELD
[0003] The present application relates to the field of medicine, and relates to use of a JAK2 inhibitor in the preparation of a medicament for treating chronic kidney disease, in particular to the use of 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 comprising the same in the preparation of a medicament for treating chronic kidney disease. BACKGROUND
[0004] Chronic kidney disease (CKD) is a kidney structural or functional abnormality caused by various reasons for more than 3 months, including the appearance of kidney injury markers (albuminuria, urinary sediment abnormalities, tubular related lesions, histological examination abnormalities and imaging examination abnormalities) or a history of kidney transplantation, with or without glomerular filtration rate (GFR) decline; or unexplained GFR decline (<60 ml / min) for more than 3 months. It is estimated that there are 850 million people worldwide with kidney disease, and the prevalence of CKD in China is 9.4% to 12.1%, with a patient population of 120 million. CKD accounts for a significant proportion of global disease burden and is still growing; about 10% of adults worldwide are affected by some form of chronic kidney disease, resulting in 1.2 million deaths each year. With the rising 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 worldwide by 2040.
[0005] Diabetic kidney disease (DKD) can be considered as one of the CKD, which is a chronic kidney disease associated with diabetic patients, and is a clinical syndrome characterized by persistent albuminuria and estimated glomerular filtration rate (eGFR) decline, which is an important cause of end-stage renal disease in China and most developed countries, and is associated with a significant increase in the incidence and mortality of cardiovascular diseases. According to the 2020 survey of the Chinese Diabetes Society, 20% to 40% of diabetic patients in China have diabetic kidney disease. If not treated in time, the kidney lesion will be irreversible, and the effect of drug treatment is poor, and the prognosis is poor.
[0006] JAK / STATs is the abbreviation of tyrosine protein kinase (JAK) and signal transduction and activation of transcription (STAT) pathway. It is widely involved in various physiological and pathological processes of cells, mediates cell reproductive differentiation, immune regulation, migration and apoptosis, and is an important signal pathway to maintain body homeostasis, which is closely related to multiple organs. As for the kidney, JAK / STATs plays an important role in primary kidney disease, secondary kidney damage, and complications of chronic kidney disease.
[0007] Currently, research targeting JAK / STATs mainly focuses on its mediation of growth factors and inflammatory factors involved in glomerular fibrosis, renal interstitial fibrosis, renal tubular epithelial cell transdifferentiation, lipid metabolism and absorption, inflammation, and other pathological processes. The application of clinical targeted therapy is less. Although there are angiotensin-converting enzyme inhibitors (ACEI), type I angiotensin II receptor blockers (ARB), sodium-glucose co-transporter 2 inhibitors (SGLT2) and other first-line therapies for the treatment of chronic kidney disease and diabetic nephropathy, the development of innovative drugs with new mechanisms and new targets still has great social significance and market prospects.
[0008] Future drug development should slow down disease progression in the 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 activation of inflammatory pathways can be found both in the early and late stages of the disease, so JAK-STATs inhibitors that inhibit important inflammatory signaling pathways, such as JAK1 inhibitors and JAK2 inhibitors, are expected to play an important role in slowing the progression of chronic kidney disease and diabetic nephropathy.
[0009] A phase II clinical trial studied the effect of barcitinib, a dual JAK1 and JAK2 inhibitor, on diabetic nephropathy, and the results showed that barcitinib can significantly reduce proteinuria in patients with diabetic nephropathy and has good clinical therapeutic effect on diabetic nephropathy.
[0010] WO2014085154A1 discloses a method of treating kidney disease. And specifically discloses a method of 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 expression level of the APOL1 polypeptide 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, lestaurtinib, ruxolitinib, CYT387, LY3009104, SD1008, cucurbitacin, GO6976, WHI-P154, AG490 and CP-690550.
[0011] In September 2021, the U.S. Food and Drug Administration (FDA) required all approved JAK inhibitors to add a black box warning to the instructions that the drugs increase the risk of serious heart-related events, cancer, blood clots, and death. By increasing the selectivity of JAK2, higher therapeutic activity can be achieved, while avoiding the side effects of JAK1 inhibitors, such as severe cytopenia, infection, secondary tumors, etc. SUMMARY
[0012] In view of the lack of JAK2 high-selective inhibitors that can inhibit important inflammatory signaling pathways in existing clinical treatment technologies for chronic kidney disease, the present application provides a use of a JAK2 inhibitor in the preparation of a drug for treating chronic kidney disease, specifically relates to the use of a compound 2-{N-[4-(4-methyl) piperazine] phenyl} amido-5, 6-dihydro-7-[3-(N-tert-butyl) sulfonamide] phenyl-pyrrolo [2, 3-d] pyrimidine or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same in the preparation of a drug for treating chronic kidney disease.
[0013] The present inventors found that the compound shown in formula 1 has high selective JAK2 inhibitory effect in previous studies. On the basis of this research and the above background, the present inventors have carried out in-depth research on the use of such compounds for the treatment of chronic kidney disease. Experimental studies have confirmed that such compounds have good therapeutic activity for chronic kidney disease caused by various reasons, can significantly reduce the level of inflammatory factors in kidney tissue, and thus reduce the degree of kidney damage and fibrosis, and have high safety, and are expected to become a new drug for treating chronic kidney disease, and have broad clinical application prospects.
[0014] The present application is realized by the following technical solutions:
[0015] The first aspect of the present application 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, the compound of formula 1 is shown as follows:
[0016] Optionally, the chronic kidney disease refers to a syndrome of decreased glomerular filtration rate caused by various chronic kidney diseases and metabolic disorders and clinical symptoms associated therewith, the renal function damage is progressive development and irreversible damage, eventually leading to complete loss of renal function; the specific diseases or conditions include primary chronic kidney disease, secondary chronic kidney disease, chronic renal tubulointerstitial lesions or genetic kidney disease.
[0017] Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, chronic nephritis evolved after acute or rapidly progressive glomerulonephritis; the secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy or lupus nephritis; the diabetic nephropathy includes chronic kidney disease associated with type 1 and type 2 diabetic patients; the chronic renal tubulointerstitial lesions include chronic pyelonephritis or chronic uric acid nephropathy.
[0018] Optionally, the pharmaceutically acceptable salt is an inorganic salt or an organic salt; the inorganic salt or the organic salt can be commonly 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.
[0019] The second aspect of the present application provides the use of a pharmaceutical composition in the preparation of a medicament for treating chronic kidney disease, characterized in that the pharmaceutical composition comprises a compound of formula 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, the compound of formula 1 is shown as follows:
[0020] Optionally, the compound of formula 1 or the pharmaceutically acceptable salt thereof in the pharmaceutical composition accounts for 0.01-90% of the mass of the pharmaceutical composition; preferably, the compound of formula 1 or the pharmaceutically acceptable salt thereof in the pharmaceutical composition accounts for 0.05-70% 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 the organic salt can be commonly 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 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 paste, sugar syrup, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, sodium alginate, and polyvinylpyrrolidone; the disintegrant is selected from one or more of crospovidone, starch, sodium bicarbonate, citric acid, tartaric acid, and low-substituted hydroxypropyl cellulose; and the solvent includes water and / or a balanced salt solution.
[0023] Optionally, the pharmaceutically acceptable carrier further includes a flavoring agent or a sweetening agent.
[0024] Optionally, the pharmaceutical composition comprising the compound of Formula 1 and pharmaceutically acceptable salts thereof is an oral preparation. Optionally, the oral preparation includes a tablet, a capsule, a granule, or an oral solution.
[0025] In another aspect, the present application 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 described above, for preventing or treating the chronic kidney disease. Optionally, the chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic tubulointerstitial disease, or genetic kidney disease.
[0026] Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, chronic nephritis evolved from acute or rapidly progressive nephritis; the secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy, or lupus nephritis; the diabetic nephropathy includes chronic kidney disease associated with type 1 or type 2 diabetes; and the chronic tubulointerstitial disease includes chronic pyelonephritis or chronic uric acid nephropathy.
[0027] The expression “effective amount” or “therapeutically effective amount” in the present application includes an amount sufficient to reduce or prevent the symptoms or conditions of a medical disease. The “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 overall health of the patient, the method, route, and dosage of administration, and the severity of side effects. The effective amount can be the maximum dose or a dose that avoids significant side effects or toxic effects.
[0028] Optionally, the therapeutically effective amount of the compound of formula 1 or its pharmaceutically acceptable salt administered is 10 mg to 1000 mg; and optionally, the therapeutically effective amount of the pharmaceutical composition administered is 10 mg to 1000 mg.
[0029] In another aspect, the present application also provides a JAK2 highly selective inhibitor for preventing or treating chronic kidney disease, wherein the chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic tubulointerstitial lesion or genetic kidney disease.
[0030] Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, chronic nephritis evolved from acute or rapidly progressive glomerulonephritis; the secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy or lupus nephritis; the diabetic nephropathy includes chronic kidney disease accompanied by type 1 or type 2 diabetes patients; and the chronic tubulointerstitial lesion includes chronic pyelonephritis or chronic uric acid nephropathy.
[0031] The present application has the following outstanding effects:
[0032] (1) The compound of formula 1 or its pharmaceutically acceptable salt as the only active ingredient of the drug can significantly reduce the level of inflammatory factors in kidney tissue, thereby reducing the degree of kidney fibrosis, and is used for treating chronic kidney disease.
[0033] (2) Compared with the first-line drugs (irbesartan and canagliflozin) currently used for treating chronic kidney disease, the compound of formula 1 has a more significant clinical advantage in terms of drug efficacy, mainly manifested as a lower onset dose and stronger treatment activity at the same dose (see Examples 1-4).
[0034] (3) Compared with the marketed JAK2 selective inhibitor fedratinib, the compound of formula 1 has a more significant advantage in terms of pharmacological activity in treating chronic kidney disease.
[0035] (4) Compared with the JAK1 and JAK2 dual-target inhibitor baricitinib which has carried out a phase II clinical study of diabetic nephropathy, the compound of formula 1 has higher selectivity for the JAK2 target. The inhibitory activity of the compound of formula 1 on JAK2 is 99 times that of the JAK1 subtype (see patent CN110305140A), while the literature reports (“Pharmacological mechanisms and clinical application research of Janus kinase inhibitors baricitinib and upadacitinib”, Chinese Journal of Clinical Pharmacology, 2023, 39(18): 2713-2717) that the IC50 of baricitinib for inhibiting JAK1 and JAK2 is 5.9 nM and 5.7 nM, respectively, without subtype selectivity. By improving the selectivity of JAK2, higher treatment activity can be obtained, and the side effects of JAK1 inhibitors, such as severe cytopenia, infection, secondary tumors, etc., can be avoided, thereby having the advantages of better activity and higher safety.
[0036] DETAILED DESCRIPTION
[0037] The application will be further described in conjunction with the specific embodiments.
[0038] Unless otherwise indicated, technical or conditions not specified in the examples were carried out according to the techniques or conditions described in the literature or according to the product instructions. The reagents or instruments used, if not specified by the manufacturer, were all conventional products available on the market.
[0039] Example 1: Study on the pharmacodynamic activity of the compound of formula 1 on the unilateral ureteral ligation (UUO) model of rats
[0040] The unilateral ureteral ligation was used to induce the renal interstitial fibrosis model in rats, and the therapeutic effect of the compound of formula 1 on renal interstitial fibrosis was investigated.
[0041] Eighty SD rats were randomly divided into 8 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, medium and high dose groups of the compound of formula 1 (2.5, 5.0, 10.0 mg / kg), 10 animals in each group. The renal interstitial fibrosis model was established by ligation of the proximal ureter of the left kidney; the sham operation group was not ligated, and the rest of the operation was the same as the model control group. The rats were given drugs once a day for 14 consecutive days. On the day after the last administration, the rats in each group were anesthetized, and blood was collected from the abdominal aorta to detect the renal function indexes (blood CRE and BUN); the left kidney tissue was dissected to detect the content of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and transforming growth factor β1 (TGF-β1) in the tissue by ELISA kit; the left kidney was dissected for HE staining and Masson staining, the degree of kidney tissue damage was observed by HE staining and scored, and the degree of kidney tissue fibrosis was observed by Masson staining and scored.
[0042] Experimental results:
[0043] 1. Effect on renal function: As shown in Table 1 below, the medium and high doses (5.0, 10.0 mg / kg) of the compound of formula 1 can significantly reduce the levels of CRE and BUN 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 levels of CRE and BUN in the blood of rats in the fedratinib group have a downward trend, but there is no statistically significant 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.
[0044] Table 1: Effect of compound of formula 1 on renal function indicators of SD rat UUO model (n=10) Note: Compared with the sham operation group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.
[0045] 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 IL-6 and TNF-α levels 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 TGF-β1 level in the renal tissue of model rats. Under 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 IL-6 and TNF-α levels of the fedratinib group have a downward trend, 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.
[0046] Table 2: Effect of compound of formula 1 on IL-6, TNF-α, TGF-β1 of SD rat UUO model (n=10) Note: Compared with the sham operation group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.
[0047] 3, Effect on morphological changes of renal tissue: As can be seen from Tables 3 and 4 below, the low, medium and high doses (2.5, 5.0, 10.0 mg / kg) of the compound of formula 1 can improve the renal tubular atrophy, renal tubular and interstitial fibrosis, renal tubular atrophy, and interstitial inflammation of the model rats, and the medium and high doses (5.0, 10.0 mg / kg) can significantly reduce the lesion score (HE staining and Masson staining) of the renal tissue of the model rats. Under 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 effect, 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.
[0048] Table 3: Effect of compound of formula 1 on HE staining pathological score of SD rat UUO model (n=10) Note: Compared with the sham operation group ++ P≤0.01; compared with the model control group ** P≤0.01, *P < 0.05.
[0049] Table 4: Effect of the compound of formula 1 on Masson staining pathology of SD rat UUO model (n = 10) Note: compared with the sham operation group ++ P < 0.01; compared with the model control group ** P < 0.01.
[0050] The above research results show that the compound of formula 1 has obvious therapeutic effect on the rat UUO model, can reduce the level of inflammatory factors in kidney tissue, and further reduce the degree of kidney fibrosis, and under 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.
[0051] Example 2: Study on the pharmacodynamic activity of the compound of formula 1 on the adriamycin-induced rat chronic kidney disease model
[0052] The adriamycin-induced rat chronic kidney disease model was used to investigate the therapeutic effect of the compound of formula 1 on chronic kidney disease. Since fedratinib did not show therapeutic activity in the rat unilateral ureteral ligation (UUO) model (no statistical difference in multiple indicators compared with the model control group), fedratinib was not used as a positive control drug in this model, and only the first-line clinical drug irbesartan was used as a positive control drug.
[0053] SD rats were injected with 3 mg / kg adriamycin via the tail vein, once a week, for 4 weeks of continuous modeling, and observed for 4 weeks to induce a rat chronic kidney disease model. Select the model successful rats, stratified random grouping according to creatinine clearance rate and body weight, respectively, as the model control group, the irbesartan group (25.0 mg / kg), the low, medium and high dose groups of the compound of formula 1 (3.0, 9.0, 27.0 mg / kg), once a day, for 8 weeks of continuous administration.
[0054] The serum urea nitrogen (BUN), creatinine (CRE) levels and urine creatinine, urine protein of each group of animals were detected before and after administration for 2, 4, 6 and 8 weeks, respectively, and the creatinine clearance rate was calculated; the right kidney tissue was dissected the day after the last administration, and the content of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and transforming growth factor-β1 (TGF-β1) in the kidney tissue was detected by rat ELISA kit, the left kidney was dissected, sectioned, embedded, and subjected to HE staining and PAS staining, respectively, to observe the morphological changes of the kidney tissue, so as to investigate the pathological changes of the kidney tissue (kidney damage degree, fibrosis degree).
[0055] Experimental results:
[0056] 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 low dose (3.0 mg / kg), and the effective time is 6 weeks after administration; the compound of Formula 1 can significantly increase the creatinine clearance rate of the model rats, the effective dose is low dose (3.0 mg / kg), and the effective time is 6 weeks after administration. Under the same dose, the activity of the compound of Formula 1 is better than that of the positive control drug (irbesartan).
[0057] Table 5: Effect of the compound of Formula 1 on renal function of SD rats at 6 weeks after administration (n = 8-10) 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.
[0058] 2. Effect on renal tissue cytokines: As shown in Table 6 below, the low, medium and high doses (3.0, 9.0, 27.0 mg / kg) of the compound of Formula 1 can significantly reduce the IL-6 and TNF-α levels of the model rats, and the high dose (27.0 mg / kg) of the compound of Formula 1 can significantly reduce the TGF-β1 level of the rats. Under the same dose, the activity of the compound of Formula 1 is better than that of the positive control drug (irbesartan).
[0059] Table 6: Effect of the compound of Formula 1 on renal tissue cytokines of SD rats (n = 8-10) Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.
[0060] 3. Effect on renal tissue: As shown in Tables 7 and 8, the 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 of the model rats, and the medium and high doses (9.0, 27.0 mg / kg) can significantly reduce the degree of renal fibrosis of the model rats and significantly reduce the PAS staining score of the kidneys of the model rats. Under the same dose, the activity of the compound of Formula 1 is better than that of the positive control drug (irbesartan).
[0061] Table 7: Effect of the compound of Formula 1 on HE staining of renal tissue of SD rats (n = 8-10) 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.
[0062] Table 8: Effect of compound of formula 1 on PAS staining score of kidney tissue of SD rats (n = 8-10) Note: compared with the normal control group ++ P < 0.01, compared with the model control group ** P < 0.01, * P < 0.05.
[0063] The above research results show that the compound of formula 1 has obvious therapeutic effect on the adriamycin-induced chronic kidney disease model in rats, and the effective dose is low dose (3.0 mg / kg), which can reduce the degree of kidney lesions and reduce the level of local tissue inflammatory factors, and under the same dose, the activity of the compound of formula 1 is better than that of the positive control drug (irbesartan).
[0064] Example 3: Study on the pharmacodynamic activity of the compound of formula 1 on the streptozotocin (STZ)-induced diabetic nephropathy model in rats
[0065] A STZ-induced type 1 diabetic nephropathy model in rats was used to investigate the therapeutic effect of the compound of formula 1 on diabetic nephropathy.
[0066] A diabetic model in rats was induced by intraperitoneal injection of 70 mg / kg STZ; the model rats were selected and randomly divided into groups according to the 24-hour urinary albumin excretion, including the model control group, the irbesartan group (25.0 mg / kg), the fedratinib group (9.0 mg / kg), the canagliflozin group (9.0 mg / kg), the low, medium and high dose groups of the compound of formula 1 (3.0, 9.0 and 27.0 mg / kg), 10 rats per group. 1 time / day, continuous administration for 8 weeks.
[0067] At the 4th, 6th and 8th weeks of administration, the urine CRE level, urinary albumin, serum BUN and CRE level were detected, and the 24-hour urinary albumin excretion, the ratio of urinary albumin / urinary CRE, the ratio of urinary CRE / serum CRE and the urinary creatinine clearance rate were calculated; the next day after the last administration, the animals in each group were euthanized under anesthesia, and the kidney tissues were dissected, the degree of kidney tissue damage was observed by HE staining and scored, and the degree of kidney tissue basement membrane sclerosis was observed by PAS staining and scored.
[0068] Experimental results:
[0069] 1. Effect on renal function: As can be seen from Table 9 below, the compound of Formula 1 can significantly reduce the ratio of urinary microalbumin to urinary CRE, increase the ratio of urinary CRE to blood CRE and creatinine clearance rate of SD rat diabetic nephropathy model, the effective dose is low dose (3.0 mg / kg), and the effective time is 6 weeks after administration. Under the same dose, the activity of the compound of Formula 1 is better than that of the positive control drugs (irbesartan, canagliflozin, fedratinib).
[0070] Table 9 Effect of the compound of Formula 1 on renal function of SD rats at 6 weeks after administration (n = 10) Note: Compared with the normal control group ++ P≤0.015; compared with the model control group ** P≤0.01, * P≤0.05.
[0071] 2. Effect on renal tissue: As can be seen from 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 significantly reduce the PAS staining score of glomerulus. Under the same dose, the activity of the compound of Formula 1 is better than that of the positive control drugs (irbesartan, canagliflozin). The fedratinib group has a certain improvement effect, but there is no statistical difference compared with the model control group.
[0072] Table 10 Effect of the compound of Formula 1 on HE staining pathological score of SD rat diabetic nephropathy model (n = 10) Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01.
[0073] Table 11 Effect of the compound of Formula 1 on PAS staining pathological score of SD rat diabetic nephropathy model (n = 10) Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.
[0074] The above research results show that the compound of Formula 1 has obvious therapeutic effect on the STZ-induced rat type 1 diabetic nephropathy model, the effective dose is low dose (3.0 mg / kg), can significantly reduce the degree of renal tissue lesions, and under the same dose, the activity of the compound of Formula 1 is better than that of the positive control drugs (irbesartan, canagliflozin, fedratinib).
[0075] Example 4: Pharmacodynamic activity study of the compound of Formula 1 on db / db mouse diabetic nephropathy model
[0076] The therapeutic effect of the compound of formula 1 on diabetic nephropathy was investigated using a spontaneous type 2 diabetic nephropathy model of db / db mice. Since fedratinib failed to show therapeutic activity on diabetic nephropathy in a streptozotocin (STZ)-induced diabetic nephropathy model of rats (no statistically significant difference in pathological score compared with the model control group), fedratinib was not used as a positive control drug in this model, and only irbesartan and canagliflozin were selected as positive control drugs.
[0077] The db / db mice were divided into groups by stratified randomization according to 24-hour urinary albumin excretion, blood glucose, and body weight, i.e., a model control group, an irbesartan group (39.0 mg / kg), a canagliflozin group (13.0 mg / kg), and a compound of formula 1 low-, medium-, and high-dose group (4.3, 13.0, and 39.0 mg / kg), 10 mice per group, once a day, for 9 weeks of continuous administration.
[0078] The 24-hour urine of each group of mice was collected at weeks 4, 6, and 9 of administration, and the urine CRE and urinary albumin concentrations were detected, and the orbital blood was collected, and the serum CRE and BUN were detected. The 24-hour urinary albumin excretion, urinary albumin / urinary CRE ratio, and urinary creatinine clearance were calculated according to the serum CRE, urine CRE, urinary albumin concentration, and 24-hour urine volume. On the day after the last administration, the right kidney cortex was taken, and the degree of kidney tissue damage was observed by HE staining and scored, and the degree of kidney tissue basement membrane sclerosis was observed by PAS staining and scored.
[0079] Experimental results:
[0080] 1. Effect on kidney function: As shown in Table 12 below, the compound of formula 1 can significantly reduce the 24-hour urinary albumin excretion of the diabetic nephropathy model of db / db mice, increase the ratio of urinary creatinine to serum creatinine, and increase the creatinine clearance rate, with a effective dose of medium dose (13.0 mg / kg) and an effective time of 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).
[0081] Table 12: Effect of the compound of formula 1 on kidney function of db / db mice at 9 weeks of administration (n = 7-10) Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.
[0082] 2. Effect on kidney tissue: As can be seen from Tables 13 and 14 below, the 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 the kidney tissue of the db / db mouse diabetic nephropathy model and significantly reduce the PAS staining score of the glomerulus, with the effective dose being the low dose (4.33 mg / kg). At the same dose, the activity of the compound of Formula 1 is superior to that of the positive control drugs (irbesartan, canagliflozin).
[0083] Table 13: Effect of the compound of Formula 1 on the HE staining pathological score of the kidney of db / db mice (n = 9-10) Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.
[0084] Table 14: Effect of the compound of Formula 1 on the PAS staining pathological score of the glomerulus of db / db mice (n = 9-10) Note: Compared with the normal control group ++ P≤0.01; compared with the model control group ** P≤0.01, * P≤0.05.
[0085] 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, with the effective dose being the low dose (4.3 mg / kg), can significantly reduce the degree of kidney tissue lesions, and at the same dose, the activity of the compound of Formula 1 is superior to that of the positive control drugs (irbesartan, canagliflozin).
[0086] The above examples only express some specific embodiments of the present application, which are described in more detail and in more detail, but should not be construed as limiting the scope of the patent of the present application. It should be noted that for those skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. Therefore, the scope of protection of the patent of the present application should be subject to the appended claims.
Claims
1. Use of a compound of Formula 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of chronic kidney disease, wherein, The compound of formula 1 is shown below:
2. Use according to claim 1, wherein, The chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic renal tubulointerstitial lesion or genetic kidney disease; Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, chronic nephritis evolved from acute or rapidly progressive glomerulonephritis; The secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy or lupus nephritis; the diabetic nephropathy includes chronic kidney disease accompanied by type 1 or type 2 diabetes mellitus patients; The chronic renal tubulointerstitial lesion includes chronic pyelonephritis or chronic uric acid nephropathy.
3. Use according to claim 1 or 2, wherein, The pharmaceutically acceptable salt is inorganic salt or organic salt; Optionally, the inorganic salt includes hydrochloride, sulfate, phosphate or nitrate; the organic salt includes methanesulfonate, citrate, oxalate or succinate.
4. Use of a pharmaceutical composition for the manufacture of a medicament for the treatment of 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, wherein the compound represented by Formula 1 is as follows:
5. Use according to claim 4, wherein, The chronic kidney disease includes primary chronic kidney disease, secondary chronic kidney disease, chronic renal tubulointerstitial lesion or genetic kidney disease; Optionally, the primary chronic kidney disease includes chronic glomerulonephritis, chronic nephritis evolved from acute or rapidly progressive glomerulonephritis; The secondary chronic kidney disease includes hypertensive nephropathy, diabetic nephropathy or lupus nephritis; the diabetic nephropathy includes chronic kidney disease accompanied by type 1 or type 2 diabetes mellitus patients; The chronic renal tubulointerstitial lesion includes chronic pyelonephritis or chronic uric acid nephropathy.
6. Use according to claim 4 or 5, characterized in that, The pharmaceutically acceptable carrier includes one or more of filler, glidant, lubricant, binder, disintegrant and solvent; Optionally, the filler is selected from one or more of mannitol, microcrystalline cellulose, starch, sucrose, dextrin, lactose, sugar powder 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 paste, sugar syrup, hydroxypropyl methyl cellulose, 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 balanced salt solution; the pharmaceutically acceptable carrier further includes flavoring agent or sweetening agent.
7. Use according to any one of claims 4-6, wherein, The pharmaceutically acceptable salt is inorganic salt or organic salt; Optionally, the inorganic salt includes hydrochloride, sulfate, phosphate or nitrate; the organic salt includes methanesulfonate, citrate, oxalate or succinate.
8. Use according to any one of claims 4-7, wherein, The dosage form of the pharmaceutical composition is oral preparation.
9. Use according to claim 8, wherein, The oral preparation includes tablet, capsule, granule or oral solution.