Application of Htr2b and inhibitor thereof in prevention or treatment of chronic kidney diseases

Abstract

The invention discloses application of Htr2b and an inhibitor thereof in prevention or treatment of chronic kidney diseases, provides application of Htr2b as a drug target in screening of drugs for prevention or treatment of chronic kidney diseases, and further provides application of the inhibitor of Htr2b in preparation of drugs for prevention or treatment of chronic kidney diseases. It is found that after the Htr2b function of DIO mice is inhibited, kidney injury caused by high fat is obviously relieved, the Htr2b inhibitor can inhibit important inflammatory factors MCP-1, IL-1, IL-6 and TNF-alpha and ectopic lipid deposition of the chronic kidney diseases, renal function markers of creatinine, urea nitrogen and UACR of the DIO mice can be obviously reduced, and the Htr2b inhibitor has the obvious effect of preventing or treating the chronic kidney diseases and has a good application prospect. The compound can be used for preparing medicines for preventing or treating chronic kidney diseases.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and relates to the application of Htr2b and its inhibitors in the prevention or treatment of chronic kidney disease, specifically the application of HTR2B and its inhibitor SB 204741 in the prevention or treatment of chronic kidney disease. Background Technology

[0002] With socioeconomic development and changes in dietary structure, the number of overweight and obese people has increased dramatically. Kidney damage caused by obesity, known as obesity-related nephropathy (ORG), is one of the main causes of end-stage renal disease. Its incidence rate is increasing in parallel with the obesity epidemic trend, and it has become a major global public health problem.

[0003] Obesity-related nephropathy (ORG) is a chronic structural and functional change in nephrons caused by obesity. Its onset is relatively insidious, with the main clinical manifestations being proteinuria, with or without hematuria, and abnormal renal function. Early ORG presents with glomerular hypertrophy and decreased glomerular density, followed by podocyte damage, tubular hypertrophy, tubulointerstitial inflammation and fibrosis, gradually progressing to FSGS. The pathogenesis of obesity-related nephropathy is complex and not fully understood, mainly involving hemodynamic changes, excessive activation of the RAAS system, abnormal secretion of adipokines, ectopic lipid deposition, and inflammation. Currently, there is no specific treatment for ORG. Treatment primarily involves a combination of measures, including weight loss, RAAS inhibitors, SGLT2 inhibitors, GLP-1 receptor agonists, and treatment of comorbid obesity, but clinical efficacy is not ideal. Therefore, the search for new drugs to treat ORG is of great significance. Summary of the Invention

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

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution.

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

[0007] Application of Htr2b inhibitors in the preparation of drugs for the prevention or treatment of chronic kidney disease.

[0008] In the above application, preferably, the inhibitor of Htr2b is SB 204741, and the structural formula of SB 204741 is shown in the following formula (1): Equation (1).

[0009] In the above-described applications, preferably, SB 204741 is used to improve ectopic lipid deposition in the kidneys.

[0010] In the above-described applications, preferably, SB 204741 is used to improve kidney inflammation.

[0011] In the above-described application, preferably, SB 204741 is applied by reducing renal function markers, which include one or more of creatinine, blood urea nitrogen, and the urine albumin / creatinine ratio.

[0012] In the above-described applications, preferably, the preparation of a medicament for the prevention or treatment of chronic kidney disease includes an inhibitor of Htr2b, a pharmaceutically acceptable salt, ester or solvent compound, and a pharmaceutically acceptable excipient.

[0013] In this invention, Htr2b is 5-hydroxytryptamine receptor 2B, i.e., 5-HT2B receptor.

[0014] In this invention, there is no special limitation on the source of the Htr2b inhibitor SB 204741. Commercially available SB 204741 commonly used in the field can be used, such as SB 204741 from MCE.

[0015] Compared with the prior art, the advantages of the present invention are as follows: This invention is the first to discover the significant effects of Htr2b inhibitors in the prevention or treatment of chronic kidney disease (CKD). Htr2b can serve as a novel target for intervention in CKD and can be used to screen drugs for its prevention or treatment. This invention found that inhibiting Htr2b function in DIO mice significantly reduced kidney damage caused by high-fat diets, indicating that Htr2b participates in and plays an important role in the process of kidney damage caused by high-fat diets. Htr2b inhibitors can inhibit important inflammatory factors in CKD such as MCP-1, IL-6, TNF-α, and ectopic lipid deposition, exhibiting significant preventive or therapeutic effects on CKD and can be used to prepare drugs for the prevention or treatment of CKD.

[0016] Based on a high-fat diet mouse model, this invention demonstrates that the Htr2b inhibitor SB 204741 significantly inhibits ectopic lipid deposition and chronic inflammation in the kidneys. It can significantly reduce renal function markers such as creatinine, blood urea nitrogen (BUN), and urine albumin / creatinine ratio (UACR) in DIO mice, thus alleviating the progression of renal CKD. Attached Figure Description

[0017] Figure 1Figure A shows the change in body weight of DIO mice and control mice over time in Example 1 of this invention. Figure B shows the appearance of the mice, Figure C shows the curve of the change in body weight of the mice over time, and Figure C shows the percentage change in body weight of the mice after drug administration.

[0018] Figure 2 This is a graph showing the daily food intake statistics of DIO mice and control mice in Example 1 of this invention.

[0019] Figure 3 This is a comparison chart of the urinary albumin / creatinine ratio (UACR) (A), creatinine (B), and blood urea nitrogen (BUN) (C) of DIO mice and control mice in Example 1 of this invention.

[0020] Figure 4 This is a comparison chart of serum total cholesterol (TC) (A) and low-density lipoprotein cholesterol (LDL) (B) in DIO mice and control mice in Example 1 of this invention.

[0021] Figure 5 This is a comparison image of HE-stained sections of kidney damage from DIO mice and control mice in Example 1 of this invention.

[0022] Figure 6 This is a comparison image of PAS-stained sections of kidney injury from DIO mice and control mice in Example 1 of this invention.

[0023] Figure 7 This is a comparison image of Oil Red O sections of the kidney cortex of DIO mice and control mice in Example 1 of this invention.

[0024] Figure 8 This is a comparison image of F4 / 80 stained kidney sections from DIO mice and control mice in Example 1 of this invention.

[0025] Figure 9 This is a graph showing the RT-PCR quantitative results of the renal inflammatory markers IL-1 (A), IL-6 (B), TNF-α (C), and MCP-1 (D) in DIO mice and control mice in Example 1 of this invention. Detailed Implementation

[0026] The present invention will be further described below with reference to the accompanying drawings and specific preferred embodiments, but this does not limit the scope of protection of the present invention. All materials and instruments used in the following embodiments are commercially available.

[0027] Example 1 The present invention relates to the application of Htr2b as a drug target in screening drugs for the prevention or treatment of chronic kidney disease.

[0028] The use of an Htr2b inhibitor of the present invention in the preparation of a medicament for the prevention or treatment of chronic kidney disease.

[0029] In this embodiment, the inhibitor of Htr2b is SB 204741, and the structural formula of SB 204741 is shown in the following formula (1): Equation (1).

[0030] In this embodiment, SB 204741 can be used to improve ectopic lipid deposition in the kidneys.

[0031] In this embodiment, SB 204741 can be used to improve kidney inflammation.

[0032] In this embodiment, the preparation of a drug for the prevention or treatment of chronic kidney disease includes an inhibitor of Htr2b, a pharmaceutically acceptable salt, ester or solvent compound, and a pharmaceutically acceptable excipient.

[0033] The following experiments demonstrate the role of the Htr2b inhibitor SB 204741 in the prevention or treatment of chronic kidney disease.

[0034] Functional study of SB 204741 in alleviating kidney damage in a DIO (Diet-Induced Obesity) mouse model.

[0035] 1. Construction of an obese mouse model SPF-grade male C57BL / 6J mice, weighing 16.71±0.55g and 5 weeks old, were randomly divided into two groups after one week of acclimatization. One group was fed a high-fat diet (HFD, Rodent Diet with 60% kcal% fat), and the other group was fed a calorie-paired control diet (NCD). The mice were weighed weekly to observe changes in body weight. The obese mouse model was considered successfully established when the degree of obesity exceeded 20%.

[0036]

[0037] HFD and NCD feed formulations are shown in Tables 1 and 2 below: Table 1. High-fat feed formulation table (HFD, No. D12492)

[0038] Table 2. Control Group Feed Ratio (NCD)

[0039] 2. Experimental grouping and drug administration 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.

[0040] 3. Detection indicators 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.

[0041] One-way ANOVA and Fisher's least significant difference (LSD) test were used to analyze the statistical significance between groups.

[0042] 4. Test Results (1) Effects of intraperitoneal injection of SB 204741 on mouse body weight and food intake.

[0043] 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.

[0044] (2) Effects of intraperitoneal injection of SB 204741 on renal function indicators in mice.

[0045] like Figure 3As 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.

[0046] (3) Effect of intraperitoneal injection of SB 204741 on the levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL) in mouse serum.

[0047] 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.

[0048] (4) Effects of intraperitoneal injection of SB 204741 on HE and PAS staining of mouse kidney pathology.

[0049] 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.

[0050] (5) Effect of intraperitoneal injection of SB 204741 on Oil Red O staining of mouse kidneys.

[0051] 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.

[0052] (6) Effect of intraperitoneal injection of SB 204741 on kidney inflammation in mice.

[0053] Figure 8 The results showed that F4 / 80 expression was low in WT and WT+SB 204741 mice, but increased in HFD mice. Treatment with SB 204741 (HFD+SB 204741 group) reduced F4 / 80 expression in mouse kidney tissue. Figure 9The results showed that the mRNA levels of IL-1, IL-6, MCP-1, and TNF-α in the kidney tissue of mice in the HFD group were increased. After treatment with SB 204741 (HFD+SB 204741 group), the levels of IL-1, IL-6, MCP-1, and TNF-α in the renal cortex were significantly lower than those in the HFD group, and the difference was statistically significant.

[0054] The above experimental results show that inhibiting Htr2b function in DIO mice significantly reduced kidney damage caused by high-fat diets, indicating that Htr2b participates in and plays an important role in the process of kidney damage caused by high-fat diets. The Htr2b inhibitor SB204741 can inhibit important inflammatory factors in chronic kidney disease, such as MCP-1, IL-6, TNF-α, and ectopic lipid deposition, and has a significant effect in preventing or treating chronic kidney disease. It can be used to prepare drugs for the prevention or treatment of chronic kidney disease.

[0055] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention using the methods and techniques disclosed above, or modify them into equivalent embodiments with equivalent changes, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solutions of the present invention shall still fall within the protection scope of the technical solutions of the present invention.

Claims

1. Application of Htr2b as a drug target in screening drugs for the prevention or treatment of chronic kidney disease.

2. Application of Htr2b inhibitors in the preparation of drugs for the prevention or treatment of chronic kidney disease.

3. The application according to claim 2, characterized in that, The inhibitor of Htr2b is SB 204741, and the structural formula of SB204741 is shown in formula (1) below: Equation (1).

4. The application according to claim 3, characterized in that, The SB 204741 is used to improve ectopic lipid deposition in the kidneys.

5. The application according to claim 3, characterized in that, The SB 204741 is used to improve kidney inflammation.

6. The application according to claim 3, characterized in that, The SB 204741 is applied by reducing renal function markers, including one or more of creatinine, blood urea nitrogen, and urine albumin / creatinine ratio.

7. The application according to any one of claims 2 to 6, characterized in that, The preparation of the medicament for the prevention or treatment of chronic kidney disease includes an inhibitor of Htr2b, a pharmaceutically acceptable salt, ester or solvent compound, and a pharmaceutically acceptable excipient.

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