Use of htr2b and inhibitors thereof in the prevention or treatment of chronic kidney disease

Abstract

The application discloses application of Htr2b and an inhibitor thereof in prevention or treatment of chronic kidney disease, proposes application of Htr2b as a drug target in screening of drugs for prevention or treatment of chronic kidney disease, and further proposes application of the inhibitor of Htr2b in preparation of drugs for prevention or treatment of chronic kidney disease. After inhibition of Htr2b function of a DIO mouse, kidney injury caused by high fat is obviously reduced, the Htr2b inhibitor can inhibit important inflammatory factors MCP-1, IL-1, IL-6, TNF-alpha and ectopic lipid deposition of chronic kidney disease, can significantly reduce kidney function markers of creatinine, urea nitrogen and UACR of the DIO mouse, has a significant effect of prevention or treatment of chronic kidney disease, and can be used for preparation of drugs for prevention or treatment of chronic kidney disease.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of biological medicine, and relates to application of Htr2b and an inhibitor thereof in prevention or treatment of chronic kidney disease, in particular to application of HTR2B and an inhibitor SB 204741 thereof in prevention or treatment of chronic kidney disease. BACKGROUND

[0002] With the development of social economy and the change of dietary structure, the number of overweight and obese people has increased dramatically. Kidney injury caused by obesity is called obesity-related glomerulopathy (ORG), which is one of the main causes of end-stage renal disease. The increasing trend of the incidence rate is parallel to the obesity epidemic trend, and has become a major global public health problem.

[0003] ORG is a chronic structural and functional change of renal units caused by obesity. The onset is relatively occult, and the main clinical manifestations are proteinuria, with or without hematuria and renal dysfunction. Glomerular hypertrophy and glomerular density decrease occur in the early stage of ORG, followed by damage to podocytes, renal tubular hypertrophy, renal tubulointerstitial inflammation and fibrosis, and gradual progression to FSGS. The pathogenesis of obesity-related glomerulopathy is complex and has not been fully elucidated, mainly including hemodynamic changes, overactivation of the RAAS system, abnormal secretion of adipokines, ectopic lipid deposition and inflammation. At present, there is no specific treatment for ORG, and comprehensive treatment measures such as weight loss, use of RAAS inhibitors, SGLT2 inhibitors and GLP-1 receptor agonists, and treatment of obesity comorbidities are mainly used, but the clinical treatment effect is not ideal. Therefore, it is of great significance to seek new drugs for the treatment of ORG. SUMMARY

[0004] The technical problem to be solved by the present application is to overcome the shortcomings of the prior art, and to provide an application of Htr2b and an inhibitor thereof in prevention or treatment of chronic kidney disease. The Htr2b inhibitor SB 204741 provided by the present application can effectively improve kidney injury, and provides a new scheme for prevention and treatment of chronic kidney disease.

[0005] To solve the above technical problems, the following technical solutions are adopted.

[0006] Application of Htr2b as a drug target in screening of drugs for prevention or treatment of chronic kidney disease.

[0007] Application of an inhibitor of Htr2b in preparation of a drug for prevention or treatment of chronic kidney disease.

[0008] The above-mentioned application is preferably that the inhibitor of Htr2b is SB 204741, and the structural formula of the SB 204741 is shown in the following formula (1):

[0009] Formula (1).

[0010] The application, preferably, the SB 204741 is applied by improving the ectopic deposition of lipids in the kidney.

[0011] The application, preferably, the SB 204741 is applied by improving the inflammation of the kidney.

[0012] The application, preferably, the SB 204741 is applied by reducing the renal function markers, including one or more of creatinine, urea nitrogen and urine albumin / creatinine ratio.

[0013] The application, preferably, the preparation of the drug for preventing or treating chronic kidney disease comprises an inhibitor of Htr2b, a pharmaceutically acceptable salt, an ester or a solvate compound and a pharmaceutically acceptable excipient.

[0014] In the present application, Htr2b is 5-hydroxytryptamine receptor 2B, i.e. 5-HT2B receptor.

[0015] In the present application, the source of the Htr2b inhibitor SB 204741 is not particularly limited, and the commercially available SB 204741 in the art can be used, such as SB 204741 from MCE company.

[0016] Compared with the prior art, the present application has the following advantages:

[0017] The present application first discovers the significant effect of the Htr2b inhibitor in preventing or treating chronic kidney disease, and Htr2b can be used as a new target for intervention of chronic kidney disease, and for screening drugs for preventing or treating chronic kidney disease. After the function of Htr2b in DIO mice is inhibited, the kidney injury caused by high fat is significantly reduced, which means that Htr2b is involved in the process of kidney injury caused by high fat and plays an important role. The Htr2b inhibitor can inhibit the important inflammatory factors MCP-1, IL-6, TNF-α and ectopic lipid deposition in chronic kidney disease, and has a significant effect on preventing or treating chronic kidney disease, and can be used for preparing drugs for preventing or treating chronic kidney disease.

[0018] Based on the high-fat diet mouse animal model, the present application finds that the Htr2b inhibitor SB 204741 has a significant inhibitory effect on the ectopic deposition of lipids in the kidney and chronic inflammation, and can significantly reduce the renal function markers of creatinine, urea nitrogen (BUN) and urine albumin / creatinine ratio (UACR) in DIO mice, and plays a role in alleviating the progression of CKD in the kidney. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1Figure 1 is a graph of the body weight of DIO mice and control mice in Example 1 of the present application over time, wherein Figure A is the appearance of the mice, Figure B is a graph of the body weight of the mice over time, and Figure C is the percentage change in body weight after the mice were dosed.

[0020] Figure 2 Figure 2 is a graph of the daily food intake of DIO mice and control mice in Example 1 of the present application.

[0021] Figure 3 Figure 3 is a comparison chart of the urinary albumin / creatinine ratio UACR (A), creatinine (B), and urea nitrogen BUN (C) of DIO mice and control mice in Example 1 of the present application.

[0022] Figure 4 Figure 4 is a comparison chart of the serum total cholesterol TC (A) and low-density lipoprotein cholesterol LDL (B) of DIO mice and control mice in Example 1 of the present application.

[0023] Figure 5 Figure 5 is a comparison chart of the kidney injury HE staining sections of DIO mice and control mice in Example 1 of the present application.

[0024] Figure 6 Figure 6 is a comparison chart of the kidney injury PAS staining sections of DIO mice and control mice in Example 1 of the present application.

[0025] Figure 7 Figure 7 is a comparison chart of the kidney cortex oil red O sections of DIO mice and control mice in Example 1 of the present application.

[0026] Figure 8 Figure 8 is a comparison chart of the kidney F4 / 80 staining sections of DIO mice and control mice in Example 1 of the present application.

[0027] Figure 9 Figure 9 is a graph of the RT-PCR quantification results of the kidney inflammation markers IL-1 (A), IL-6 (B), TNF-a (C), and MCP-1 (D) of DIO mice and control mice in Example 1 of the present application. DETAILED DESCRIPTION

[0028] The present application is further described below in conjunction with the accompanying drawings and specific preferred embodiments, but the scope of the present application is not limited by the same. The materials and instruments used in the following examples are commercially available.

[0029] Example 1

[0030] An application of Htr2b of the present application as a drug target in screening drugs for preventing or treating chronic kidney disease.

[0031] Use of an inhibitor of Htr2b of the present application in the manufacture of a medicament for preventing or treating chronic kidney disease.

[0032] In this embodiment, the inhibitor of Htr2b is SB 204741, and the structure of SB 204741 is shown in the following formula (1):

[0033] Formula (1).

[0034] In this embodiment, SB 204741 can be used by improving ectopic deposition of renal lipids.

[0035] In this embodiment, SB 204741 can be used by improving inflammation of the kidney.

[0036] In this embodiment, the medicament for preventing or treating chronic kidney disease comprises an inhibitor of Htr2b, a pharmaceutically acceptable salt, ester or solvate compound, and a pharmaceutically acceptable excipient.

[0037] The following experiments demonstrate the effect of the inhibitor of Htr2b SB 204741 on preventing or treating chronic kidney disease.

[0038] Functional study of SB 204741 in reducing kidney damage in a DIO (Diet-Induced Obesity) mouse model.

[0039] 1. Construction of an obese mouse model

[0040] SPF grade C57BL / 6J male mice, weighing 16.71±0.55g, 5 weeks old, were randomly adapted to feed for one week, and then randomly divided into two groups, fed with high-fat diet (HFD, Rodent Diet with 60% kcal% fat) and calorie-matched control diet (NCD), respectively. The body weight of the mice was measured every week, and the change of the body weight of the mice was observed. When the obesity degree was more than 20%, the obese mouse model was successfully established.

[0041]

[0042] The HFD and NCD feed formulations are shown in Tables 1 and 2 below:

[0043] Table 1 High-fat feed formulation table (HFD, No. D12492)

[0044]

[0045] Table 2 Control group feed formulation table (NCD)

[0046]

[0047] 2. Experimental grouping and drug administration

[0048] After 22 weeks of continuous modeling, mice on a normal diet were randomly divided into two groups (WT, WT+SB 204741) and mice successfully modeled as obese were randomly divided into two groups (HFD, HFD+SB 204741). Significant differences in body weight were ensured between the two groups on the normal diet and between the two groups on the high-fat diet. After grouping, the experimental period lasted 3 weeks. The HFD and HFD+SB 204741 groups continued to be fed a high-fat diet, while the WT and WT+SB 204741 groups continued to be fed a basal diet. Food intake was recorded daily. The WT and HFD groups received intraperitoneal injections of 5% DMSO at a dose of 5 mg / kg / day every morning, while the WT+SB 204741 and HFD+SB 204741 groups received intraperitoneal injections of SB 204741 at a dose of 5 mg / kg / day every morning.

[0049] 3. Detection indicators

[0050] After the experimental period ended, mice were fasted for at least 12 hours the day before sampling, with no restriction on water intake during the fasting period. Before euthanasia, the mice were weighed and their weight recorded. Mice were then intraperitoneally injected with the appropriate volume of 3% pentobarbital anesthetic based on their body weight. After anesthesia, blood was collected from the eyeballs in 1.5 mL anticoagulant EP tubes, incubated at room temperature for 3 hours, centrifuged at 3000 rpm for 10 minutes at room temperature, and the supernatant was carefully aspirated, aliquoted, and frozen at -80°C for later analysis. Tissue specimens: Mouse kidney tissue was isolated, placed in pre-cooled physiological saline, and blood and impurities were removed. The tissue was blotted dry with filter paper, weighed, and its weight recorded. The tissue was stored at -80°C for later use.

[0051] One-way ANOVA and Fisher's least significant difference (LSD) test were used to analyze the statistical significance between groups.

[0052] 4. Test Results

[0053] (1) Effects of intraperitoneal injection of SB 204741 on mouse body weight and food intake.

[0054] like Figure 1 As shown in the AC diagram, after 22 weeks of HFD feeding, the body weight of mice in the HFD group was significantly higher than that in the WT group (p < 0.001). After 3 weeks of intervention with SB 204741, the weight gain trend of the treated group slowed down, and the body weight of the mice was lower than that of the HFD group after three weeks of treatment, indicating that SB 204741 has a significant weight-loss effect. Figure 2 As shown, the mice's food and energy intake were unaffected.

[0055] (2) Effects of intraperitoneal injection of SB 204741 on renal function indicators in mice.

[0056] like Figure 3 As shown in the AC diagram, compared with the WT group, the serum creatinine and blood urea nitrogen (BUN) levels of mice in the HFD group were significantly increased (p < 0.001). After 3 weeks of intervention with SB 204741, the serum creatinine and BUN levels of mice in the HFD+SB 204741 group were lower than those in the HFD group, but there was no statistically significant difference compared with the WT group. Similarly, compared with the WT group, the urinary UACR level of mice in the HFD group was significantly increased. After intervention with SB 204741, the urinary UACR level of mice in the HFD+SB204741 group was lower than that in the HFD group, but there was no statistically significant difference compared with the WT group.

[0057] (3) Effect of intraperitoneal injection of SB 204741 on the levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL) in mouse serum.

[0058] Figure 4 The results of the AB study showed that, compared with the WT group, the serum TC and LDL levels of mice in the HFD group were increased (p<0.001), while the serum TC and LDL levels of mice in the SB 204741 group were significantly reduced (p<0.05), indicating that SB 204741 can alleviate lipid accumulation in DIO mice.

[0059] (4) Effects of intraperitoneal injection of SB 204741 on HE and PAS staining of mouse kidney pathology.

[0060] according to Figure 5 and Figure 6 It was observed that HFD mice exhibited glomerular hypertrophy, mesangial proliferation, and tubular dilatation with exposed nuclei and casts in the kidneys under HE and PAS staining. Intraperitoneal injection of SB 204741 could partially alleviate these symptoms.

[0061] (5) Effect of intraperitoneal injection of SB 204741 on Oil Red O staining of mouse kidneys.

[0062] according to Figure 7 It was found that intraperitoneal injection of SB 204741 significantly reduced lipid deposition in the glomeruli and renal tubules, as observed under Oil Red O staining of the kidneys.

[0063] (6) Effect of intraperitoneal injection of SB 204741 on kidney inflammation in mice.

[0064] Figure 8F4 / 80 expression was less in WT and WT+SB 204741 groups, and increased in HFD group, and SB 204741 treatment (HFD+SB 204741 group) reduced F4 / 80 expression in kidney tissue of mice. Figure 9 Figures 5A-D show that IL-1, IL-6, MCP-1, and TNF-α mRNA levels in kidney tissue of HFD group mice increased, and after SB 204741 treatment (HFD+SB 204741 group), IL-1, IL-6, MCP-1, and TNF-α levels in kidney cortex were significantly lower than in HFD group, and the difference was statistically significant.

[0065] From the above test results, it can be seen that after inhibiting the function of Htr2b in DIO mice, the kidney injury caused by high fat is significantly reduced, and Htr2b participates in the process of kidney injury caused by high fat and plays an important role. Htr2b inhibitor SB204741 can inhibit important inflammatory factors MCP-1, IL-6, TNF-α and ectopic lipid deposition in chronic kidney disease, and has a significant effect on preventing or treating chronic kidney disease, and can be used for preparing a drug for preventing or treating chronic kidney disease.

[0066] The above is only a preferred embodiment of the present application, and does not limit the present application in any form. Although the present application has been disclosed as above with a preferred embodiment, it is not intended to limit the present application. Any person skilled in the art can make many possible changes and modifications to the technical solutions of the present application, or modify equivalent embodiments, without departing from the spirit and technical solutions of the present application. Therefore, any simple modification, equivalent replacement, equivalent change and modification of the above embodiments according to the technical essence of the present application, which does not deviate from the technical solutions of the present application, still belongs to the protection scope of the technical solutions of the present application.

Claims

1. Use of an inhibitor of Htr2b for the manufacture of a medicament for the prevention or treatment of chronic kidney disease, characterized in that, The chronic kidney disease is chronic obesity-related kidney disease, the inhibitor of Htr2b is SB 204741, and the structural formula of SB 204741 is shown in the following formula (1): Formula (1).

2. Use according to claim 1, characterized in that, The SB 204741 is applied by improving ectopic deposition of renal lipids.

3. Use according to claim 1, characterized in that, The SB 204741 is applied by improving inflammation of the kidney.

4. Use according to claim 1, characterized in that, The SB 204741 is applied by reducing a renal function marker, and the renal function marker includes one or more of creatinine, urea nitrogen and urine albumin / creatinine ratio.

5. The use according to any one of claims 1 to 4, characterized in that, The preparation of the drug for preventing or treating chronic kidney disease includes an inhibitor of Htr2b, a pharmaceutically acceptable salt, an ester or a solvate compound and a pharmaceutically acceptable adjuvant.

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