Use of diosmin as a drug for improving glucose and lipid metabolism disorders

Abstract

The present invention relates to the use of diosmin as a drug for preparing a drug for improving glucose and lipid metabolism disorders. The present invention provides a new drug for improving glucose and lipid metabolism disorders, especially lipid metabolism disorders, overcomes the deficiencies of existing treatment methods, overcomes the side effects caused by existing thiazolidinediones, and opens up more options for clinical treatment of drugs for improving glucose and lipid metabolism disorders.

Description

Technical Field

[0001] The present invention relates to a new use of diosmin, in particular to the use of diosmin as a medicine for preparing a drug for improving glucose and lipid metabolism disorders. Background Art

[0002] Obesity and diabetes are a global health problem, and the development of therapeutic drugs for them has always been a hot topic of research. Thiazolidinediones (TZDs) are the only insulin sensitizers currently used in clinical practice. They have a definite therapeutic effect, but numerous reports of adverse reactions have hindered the continued use of this class of drugs. Since their launch in 1999, thiazolidinediones, represented by rosiglitazone, have been considered to be the best insulin sensitizers in clinical practice and have long been used as first-line drugs. However, with the widespread clinical application, a series of adverse reactions of this class of drugs have been reported, such as weight gain, water and sodium retention, osteoporosis, tumors and cardiovascular risks. In particular, long-term medication-induced cardiovascular events have attracted widespread attention, which has directly led to the suspension or restriction of the use of this class of drugs. However, there are no better alternatives to other insulin sensitizers in clinical practice, which has brought new challenges and opportunities to the drug development of new insulin sensitizers. Peroxisome proliferator-activated receptor γ (PPARγ) is a known receptor for TZDs, and its pharmacodynamic effects and adverse reactions have been studied in depth. In order to retain the insulin sensitizing effect mediated by PPARγ to the greatest extent and minimize the occurrence of related adverse reactions, the concept of "selective PPARγ modulators (SPPARMs)" has been proposed and developed to guide the practice of new drug research and development.

[0003] At present, the PPARγ targeted compounds under development internationally can be divided into the following categories: partial agonists, PPARγ phosphorylation inhibitors and TZDs derivatives. Selective PPARγ modulators have similar or different PPARγ receptor binding sites, or have different affinities or specificities for recruiting receptor cofactors, or have tendencies and selectivity for a series of target genes that PPARγ regulates transcription. However, the activity regulation of PPARγ has a complex regulatory mechanism, involving PPARγ-mediated gene transcription regulation, PPARγ protein post-translational modification, and other non-PPARγ proteins and their mechanisms of action.

[0004] Taking partial agonists as an example, when screening existing drugs, rosiglitazone was used as a reference, and the dose-effect of in vitro transcriptional activation of PPARγ and the binding force with PPARγ receptors were experimentally screened to preliminarily screen out compounds that may serve as partial agonists. At present, academics believe that drugs that achieve 20-60% of the efficacy of rosiglitazone may serve as partial agonists, but not all agonists with reduced efficacy can achieve insulin sensitization and have less or no TZD adverse reactions. The binding sites and binding regions of partial agonists and PPARγ determine the conformation of PPARγ and the subsequent changes in cofactor recruitment. The selective recruitment of cofactors determines the selectivity of PPARγ in regulating target genes.

[0005] Since the mechanism of action of selective PPARγ modulators in regulating PPARγ activity is currently unclear, the research focus in recent years on the development of new therapeutic drugs to improve glucose and lipid metabolism has been to initially screen out drugs that may act as partial agonists from existing drugs and then look for a selective PPARγ modulator compound that improves glucose and lipid metabolism without causing obvious side effects. Summary of the invention

[0006] In view of the deficiencies in the prior art, the purpose of the present invention is to find a new drug for improving glucose and lipid metabolism disorders, overcome the deficiencies of existing treatment methods, overcome the side effects caused by existing thiazolidinediones, and open up more options for clinical treatment of glucose and lipid metabolism disorders.

[0007] To achieve the above object, the present invention provides the use of diosmin as a drug for preparing a drug for improving glucose and lipid metabolism disorders.

[0008] Furthermore, the glucose and lipid metabolism disorder includes lipid metabolism disorder.

[0009] Furthermore, the drug is a drug for preventing and / or treating obesity.

[0010] Furthermore, the drug is a drug for inhibiting fat accumulation.

[0011] Furthermore, the drug is a drug that increases basal metabolic rate / resting energy expenditure.

[0012] Furthermore, the drug is a lipid-lowering drug.

[0013] Furthermore, the drug is a partial PPARγ agonist.

[0014] In addition, the present invention also provides the use of diosmin as a drug for preparing a drug for preventing and / or treating atherosclerosis, coronary heart disease, stroke and peripheral arterial disease caused by elevated low-density lipoprotein levels.

[0015] The present invention has the following advantages:

[0016] (1) Compared with insulin sensitizers of the thiazolidinedione class of drugs represented by rosiglitazone, diosmin in the present invention has a weaker activation effect on PPARγ transcription. It is found that it can be used as a partial agonist of PPARγ without the side effects of insulin sensitizers of the thiazolidinedione class of drugs.

[0017] (2) Diosmin in the present invention can effectively inhibit adipocyte differentiation and can be used to prepare drugs for preventing and / or treating diseases related to glucose and lipid metabolism disorders such as obesity and hyperlipidemia.

[0018] (3) The diosmin in the present invention can inhibit the production of lipid droplets in the cytoplasm of adipocytes and can be used to prepare drugs for preventing and / or treating diseases related to glucose and lipid metabolism disorders such as obesity and hyperlipidemia.

[0019] (4) The present invention has more comprehensively explored the medicinal value of diosmin and expanded its clinical application, providing a new source and direction for the development of active ingredients for drugs for treating diseases related to glucose and lipid metabolism disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Agonist efficacy of diosmin and rosiglitazone on PPARγ.

[0021] Figure 2 a Comparison of insulin sensitivity between high-fat-fed mice in the diosmin group and the control group; b Body weight changes of high-fat-fed mice in the diosmin group and the control group; c Changes in body fat content of high-fat-fed mice in the diosmin group and the control group; d Comparison of tissue weights of subcutaneous adipose tissue iWAT and visceral adipose tissue eWAT in high-fat-fed mice in the diosmin group and the control group.

[0022] Figure 3 a Comparison of the amount of lipid droplets in brown adipose tissue (BAT), subcutaneous adipose tissue (iWAT) and visceral adipose tissue (eWAT) of high-fat-fed mice in the diosmin group and the control group; b Comparison of the average area of ​​lipid droplets in subcutaneous adipose tissue (iWAT) and visceral adipose tissue (eWAT) of high-fat-fed mice in the diosmin group and the control group.

[0023] Figure 4 Comparison of low-density lipoprotein levels in high-fat-fed mice treated with diosmin and the control group.

[0024] Figure 5 a Comparison of PGC1α and UCP1 thermogenic proteins in C57 / BL6J mice between the diosmin group and the control group; b Comparison of oxygen consumption and carbon dioxide production in high-fat-fed mice between the diosmin group and the control group; c Comparison of activity levels in high-fat-fed mice between the diosmin group and the control group; d Comparison of food intake in high-fat-fed mice between the diosmin group and the control group.

[0025] Figure 6 a Comparison of water content between high-fat-fed mice in the diosmin group and control group; b Comparison of heart weight between high-fat-fed mice in the diosmin group and control group; c Comparison of subcutaneous heart sections between high-fat-fed mice in the diosmin group and control group; d Comparison of M-mode echocardiograms between high-fat-fed mice in the diosmin group and control group; e Comparison of cardiac ejection fraction between high-fat-fed mice in the diosmin group and control group; f Comparison of ventricular short-axis shortening rate between high-fat-fed mice in the diosmin group and control group; g Comparison of heart wall thickness between high-fat-fed mice in the diosmin group and control group; h Comparison of left ventricular end-systolic diameter between high-fat-fed mice in the diosmin group and control group; i Expression levels of genes related to myocardial hypertrophy and myocardial fibrosis in myocardial tissue of high-fat-fed mice in the diosmin group and control group. DETAILED DESCRIPTION

[0026] The present invention will be further described in detail below with reference to embodiments and effect examples, but the scope of the present invention is not limited thereto.

[0027] Example 1: Screening of compounds with strong binding ability to PPARγ by computer model

[0028] The crystal structure of the human PPARγ ligand binding domain (PDB code 5GTO) was retrieved from the RCSB protein database and imported into the SYBYL software (Tripos, St, Louis, MO) for subsequent structural analysis and docking simulation. The ligand database contains 3000 compounds from the Targetmol Bioactive Natural Compound Library (https: / / www.tsbiochem.com). After the PPARγ protein and candidate ligands were prepared, molecular docking was performed using the Surflex-Dock module of SYBYL. Using "binding energy" as an indicator, four compounds with strong binding energy were obtained. Among them, diosmin with the strongest binding energy was selected to further study its improvement effect on glucose and lipid metabolism at the in vitro and in vivo levels.

[0029] Example 2: PPARγ agonist efficacy of diosmin

[0030] In order to detect the agonistic ability of diosmin on PPARγ, its agonistic ability was compared with that of the full agonist rosiglitazone, and a luciferase reporter experiment was performed. The PPRE-promoter-luciferase reporter plasmid was transfected into HEK-293 cells, and HEK-293 cells were divided into diosmin group and rosiglitazone group, and 0.01μM, 0.1μM, 1μM, and 10μM diosmin and rosiglitazone were added respectively to detect the effect of diosmin on PPARγ transcriptional activity. The results showed that compared with rosiglitazone, the activation effect of diosmin on PPARγ transcription was weaker with the increase of concentration. When 0.1μM diosmin and rosiglitazone were added, its efficacy was only less than 50% of rosiglitazone, and diosmin was verified to be a partial agonist of PPARγ.

[0031] Example 3: Diosmin improves insulin sensitivity and resists obesity

[0032] Establishment of HFD mouse model: The experimental mice were 6-week-old C57BL / 6 mice raised in the laboratory of Wenzhou Medical University. The mice were fed a high-fat diet for 12 weeks.

[0033] The HFD mice were divided into two groups, one group was the control group, and the other group was the diosmin group. The diosmin group was injected with 10 mg / kg of diosmin compound into the mouse subcutaneous adipose tissue (iWAT) every week, and the control group was injected with an equal volume of solvent (olive oil) into the mouse iWAT tissue every week; and the insulin tolerance (ITT) was tested at the eleventh and twelfth week. The results are as follows Figure 2 As shown in a, compared with the control group, the insulin sensitivity of mice treated with diosmin was significantly improved. Through body weight testing, it was found that diosmin treatment could significantly reduce the body weight of HFD mice. Figure 2 b. After 12 weeks of high-fat diet, body composition analysis was performed. The results showed that the body fat content of the diosmin-treated group was significantly reduced compared with the control group. Figure 2 c. The effect of diosmin on lipid accumulation in various adipose tissues was also tested. By weighing the adipose tissue, it was found that the weight of subcutaneous adipose tissue iWAT and visceral adipose tissue eWAT was significantly reduced after diosmin treatment. Figure 2 In summary, diosmin improves high-fat diet-induced obesity and insulin resistance by increasing insulin sensitivity.

[0034] Example 4: Diosmin inhibits fat accumulation

[0035] The adipose tissues of the HFD mice in the diosmin group and the HFD mice in the control group were sliced. Figure 3a shows that after diosmin treatment, the fat droplets in the brown adipose tissue (BAT) of mice decreased, the fat droplets in iWAT and eWAT decreased significantly, and the number of adipocytes increased. To confirm this change, the iWAT and eWAT sections were manually quantified using Image Pro Plus software, and it was found that the average area of ​​fat droplets in iWAT and eWAT of mice treated with diosmin was significantly reduced. The results are shown in Figure 3 As shown in b. This shows that diosmin can effectively inhibit fat accumulation in obese mice.

[0036] Example 5: Diosmin improves blood lipids and reduces low-density lipoprotein expression levels

[0037] After 12 weeks of high-fat feeding, serum was collected from the diosmin group and the control group, and blood lipid indicators were tested. It was found that the low-density lipoprotein level of mice was significantly reduced after diosmin treatment. Figure 4 It shows that diosmin can improve blood lipid indexes, especially low-density lipoprotein levels, in patients with lipid metabolism disorders, which suggests that diosmin can be used as a drug to prevent atherosclerosis, coronary heart disease, stroke and peripheral arterial disease caused by elevated low-density lipoprotein.

[0038] Example 6: Diosmin enhances energy expenditure

[0039] Diosmin was subcutaneously injected at a dose of 10 mg / kg in the groin of C57 / BL6J mice, and the mice were killed 3 days later. It was found that diosmin treatment promoted the expression of thermogenic proteins such as PGC1α and UCP1. Figure 5 As shown in a. In order to explore whether diosmin affects the energy consumption of mice, C57 / BL6J mice were placed in metabolic cages (Columbus Instrument, USA) to monitor the energy consumption of C57 / BL6J mice. Through real-time monitoring in the metabolic cage, it was found that the oxygen consumption and carbon dioxide production of C57 / BL6J mice in the diosmin-treated group were significantly higher than those in the control group. The results are shown in Figure 5 As shown in Figure b, this reflects that diosmin does cause an increase in energy consumption in mice. In addition, there was no significant difference in the activity and food intake of the two groups of mice during the experiment, indicating that the increase in energy consumption was not caused by these factors. Diosmin increased the energy consumption of obese mice by promoting thermogenesis in obese mice.

[0040] Example 7: Diosmin does not have the side effects associated with thiazolidinediones

[0041] In the present invention, the common side effects of TZD drugs were detected. C57 / BL6J mice were divided into a diosmin group and a control group. The diosmin group was injected subcutaneously with 10 mg / kg diosmin in the groin of C57 / BL6J mice, and the control group was injected with an equal volume of solvent (olive oil) in the groin of C57 / BL6J mice. The mice were killed after 12 weeks, and the water content of the mice was first analyzed by a body composition analyzer. The results are as follows: Figure 6 As shown in a, it was found that diosmin treatment did not cause fluid retention. Compared with the control group, the heart weight of mice in the diosmin-treated group did not increase. Figure 6 As shown in b, the heart sections also showed no myocardial hypertrophy or cardiac chamber enlargement. Figure 6 c. M-mode echocardiography also showed no obvious abnormalities. Figure 6 d. Consistent with this, diosmin treatment did not affect the cardiac function of mice. The results of mouse cardiac ultrasound examination showed that there was no significant difference between the diosmin treatment group and the control group in terms of cardiac ejection fraction, ventricular short axis shortening rate, heart wall thickness, and left ventricular end-systolic diameter ( Figure 6 eh). Overload of the heart can cause myocardial hypertrophy and myocardial fibrosis, leading to cardiac remodeling. Therefore, the expression levels of genes Anp, Bnp, β-Mhc, Acta1, Rcan1.4, cTGF, Col1a1, and Col3a1 related to myocardial hypertrophy and myocardial fibrosis in myocardial tissue were detected again, and it was found that there was no significant difference in gene expression between the two groups of mice ( Figure 6 i). In summary, diosmin, as a novel selective PPARγ modulator, improves glucose and lipid metabolism without causing obvious side effects associated with thiazolidinediones in the prior art.

[0042] Finally, it is necessary to explain here that the above embodiments are only used to further illustrate the technical solution of the present invention in detail and cannot be understood as limiting the scope of protection of the present invention. Some non-essential improvements and adjustments made by technicians in this field based on the above content of the present invention all fall within the scope of protection of the present invention.

Claims

1. Use of diosmin for preparing a drug for increasing insulin sensitivity.

2. The use according to claim 1, characterized in that the drug is a partial PPARγ agonist.

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