Drug application of 2-myricetin-based diselenide in atherosclerosis aspect

Abstract

The invention discloses application of two-myricetin-based diselenide in preparation of a pharmaceutical composition, which is used for preventing, relieving and / or treating cardiovascular diseases and preventing and / or treating atherosclerosis.

Description

Technical Field

[0001] The present application relates to the field of medical technology, and in particular to a pharmaceutical application of dimyricetin-based diselenide. Background Art

[0002] Due to the aging of the population, changes in lifestyle, and the continued prevalence of metabolic risk factors, the prevalence of cardiovascular diseases has increased year by year worldwide. In my country, due to the large population base, the health and economic burden brought by cardiovascular diseases is particularly serious. In 2019, cardiovascular diseases accounted for 46.74% and 44.26% of the causes of death in rural and urban areas in my country, and 2 out of every 5 deaths were caused by cardiovascular diseases. Among cardiovascular diseases, coronary atherosclerotic heart disease (CHD) is one of the diseases with the highest morbidity and mortality. Although with the gradual popularization of interventional treatment and the development of lipid-lowering drugs such as statins, intervention measures have been taken for conventional high-risk factors for atherosclerosis such as hyperlipidemia, smoking, hypertension, and diabetes. The disease progression of patients with coronary heart disease has been curbed, and the risk of death has also decreased, but CHD is still one of the main causes of death among patients worldwide.

[0003] Atherosclerosis (AS) is the main cause of coronary heart disease, cerebral infarction, and peripheral vascular disease. Lipid metabolism disorders are the pathological basis of atherosclerosis, and its characteristic is that the lesions of the affected arteries start from the intima, generally with lipid and complex carbohydrate accumulation, bleeding and thrombosis, followed by fibrosis and calcification, and gradual degeneration and calcification of the middle layer of the artery, leading to thickening and hardening of the arterial wall and narrowing of the blood vessel lumen. The lesions often involve large and medium muscular arteries. Once they develop to the point of blocking the arterial lumen, the tissues or organs supplied by the artery will be ischemic or necrotic.

[0004] Selenium is an essential trace element for the human body. The Chinese Nutrition Society also lists selenium as one of the 15 essential nutrients for the human body. A large number of clinical experiments at home and abroad have shown that selenium deficiency in the human body can cause dysfunction of certain important organs and lead to many serious diseases. Studies have shown that people with low selenium or selenium deficiency can not only prevent the occurrence of tumors, liver diseases, etc. by taking appropriate amounts of selenium supplements, but also improve the body's immune ability, maintain the normal functions of important organs such as the heart, liver, lungs, and stomach, and prevent the occurrence of cardiovascular and cerebrovascular diseases in the elderly.

[0005] Myricetin is a flavonoid compound that is widely found in the bark and leaves of bayberry trees. It has multiple pharmacological activities such as anti-inflammatory, anti-tumor, anti-mutation, caries prevention, antioxidant properties, and the ability to eliminate free radicals in the body.

[0006] Diselenides have important antioxidant effects and can simulate the activity of glutathione peroxidase (GSH-PX). Some diselenides also have anti-tumor, antibacterial, bactericidal and disinfectant activities.

[0007] Dimyricetin-yl-diselenide (DMS), molecular formula: C 30 H 18 O 16 Se 2 , molecular weight: 792.37, properties: light yellow powder, it is a new compound discovered by our company. Summary of the invention

[0008] The present application provides a pharmaceutical application of myricetin-based diselenide.

[0009] The present application adopts the following technical scheme: the use of myricetin diselenide for preparing a pharmaceutical composition for preventing, alleviating and / or treating cardiovascular diseases, and for preventing and / or treating atherosclerosis.

[0010] The beneficial effects of this application are as follows:

[0011] Experiments have shown that myricetin-based diselenide is useful for preventing and / or treating atherosclerosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0013] Figure 1 The effect of the drug to be tested on the body weight of mice.

[0014] Figure 2 ApoE - / - Effects of serum lipids and plasma LDH levels in mice.

[0015] Figure 3 ApoE - / - Effects of plasma SOD and MDA on mice.

[0016] Figure 4 ApoE - / - Effects of aortic plaque in mice.

[0017] Figure 5 ApoE - / - Effects of aortic plaque in mice.

[0018] Figure 6 ApoE - / - Effect of cardiac weight index on mice. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical solution and advantages of the present application clearer, the technical solution of the present application will be clearly and completely described below in combination with the specific embodiments of the present application and the corresponding drawings. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present application.

[0020] Example: Study on the efficacy of the drug on the atherosclerosis model of ApoE- / - mice fed with high fat diet

[0021] 1. Experimental Animals

[0022] 1.1 Experimental animals

[0023] ApoE - / - Mice (ApoE knockout mice) were 56-62 days old, male, SPF grade, provided by Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. License number: SCXK (Beijing) 2021-0006, certificate number: No.110011230100310621.

[0024] 1.2 Rearing environment

[0025] All animals used in the experiment were ApoE - / - Mice, after arriving at Nantong WuXi AppTec Pharmaceutical Technology Co., Ltd., were kept in an SPF-level animal breeding room with strictly controlled environmental conditions. The temperature in the breeding room was maintained at 20-25°C and the humidity was maintained at 40-70%. The temperature and humidity in the breeding room were monitored in real time by a thermometer and hygrometer, and the temperature and humidity were recorded twice a day (once in the morning and once in the afternoon). The lighting in the animal breeding room was controlled by an electronic timed lighting system, with the lights on for 12 hours and off for 12 hours a day (6:00AM on, 18:00PM off). Animals were free to eat feed and drink water.

[0026] 2. Experimental Methods

[0027] 2.1 Experimental Materials and Instruments

[0028] 2.1.1 Reagents and suppliers

[0029] Reagents supplier Production batch number TRIG Kit Beckman AUZ 0735 CHOL Kit Beckman AUZ 0608 HDL Kit Beckman AUZ 1158 LDL Test Kit Beckman AUZ 1232 LDH Kit Beckman AUZ 0982 SOD Kit BioSky Biotech Co., Ltd. 030223230412 MDA Kit Beijing Prilai Gene Technology Co., Ltd. 2023R2G 0.9% Sodium Chloride Injection Huayu (Wuxi) Pharmaceutical Co., Ltd. 20011402 DMSO RK8RRQ1Q Anhui Zesheng Technology Co., Ltd. PEG400 R20120011 Shanghai Shaoyuan Reagent Co., Ltd. Glacial acetic acid Sane Chemical Technology (Shanghai) Co., Ltd. DX3REA9K Oil-red O Sigma SLCG5388 Isopropyl alcohol Sinopharm Chemical Reagent Co., Ltd. 20210605 4% paraformaldehyde Shanghai Bio-Tech Co., Ltd. 112922230324

[0030] 2.1.2 Instruments and suppliers

[0031]

[0032]

[0033] 2.2 Experimental Procedure

[0034] ApoE - / - The mice were acclimatized to the experimental facility for 3 days after arrival. After fasting overnight (12 hours), blood was collected to measure blood lipids TC, TG, HDL-C, LDL-C and LDH as the basic values. The control mice were fed with regular diet, and the other mice were fed with high-fat diet for a total of 18 weeks. The regular diet contained 11% kal fat and 0% cholesterol; the high-fat diet contained 40% kal fat and 0.21% cholesterol; after 8 weeks of high-fat feeding, the animals underwent cardiac ultrasound and blood biochemistry tests, and were randomly divided into blank control, model control, positive control, AQ-Ca, DMS-ASD and DMS groups according to TC values ​​(see Table 2 in 6.5). The drug administration lasted for 10 weeks. During the experiment, the appearance and behavior of the animals were observed, and body weight, cardiac ultrasound, blood lipids (TC, TG, HDL-C and LDL-C) and blood biochemistry (LDH, SOD and MDA) were measured. At the end of the experiment (week 18), the heart and aorta were collected for oil red staining and histopathological analysis.

[0035] 2.3 Test article and high-fat diet

[0036]

[0037] 2.4 Drug configuration

[0038] Solvent 1: 5% DMSO + 95% DD water;

[0039] Solvent 2: 20% DMSO+60% PEG400+20% 10mM Acetic buffer (pH4.5)

[0040] 1) Weigh an appropriate amount of AQ-Ca, add it to 5% DMSO, vortex for 1 minute, dilute with water to the target concentration, vortex for 1 minute for later use. The solution is a suspension and should be used immediately after preparation;

[0041] 2) Weigh an appropriate amount of DMS-ASD, add it to 5% DMSO, vortex for 1 minute, dilute with water to the target concentration, vortex for 1 minute for later use. The solution is a suspension and should be used immediately after preparation;

[0042] 3) Weigh an appropriate amount of DMS, determine the final volume based on the concentration, add to 20% DMSO, vortex for 1 min, add 60% PEG400, vortex for 1 min, add 20% 10mM Acetic buffer (pH4.5), vortex for 1 min, and use it immediately.

[0043] Storage: Prepare the compound on the day of administration and store at 4°C. Take it out of the refrigerator when using.

[0044] 2.5 Experimental Grouping

[0045] Group plan quantity Dosage (mg / kg) Route of administration Schedule 1 Regular diet+Vehicle 8 - PO QD 2 HFD+Vehicle 6 - PO QD 3 HFD+atorvastatin 6 20 PO QD 4 HFD+AQ-Ca 6 300 PO QD 5 HFD+DMS-ASD 6 300 PO QD 6 HFD+DMS 8 20 SC QD

[0046] 2.6 Endpoint detection

[0047] 2.6.1 Weight

[0048] All animals were treated once a week and once a day during the dosing period;

[0049] 2.6.2 Blood biochemical test

[0050] The animals were fasted the night before blood collection, blood was collected, heparin was used for anticoagulation, and plasma was prepared for testing TG, TC, HDL-C, LDL-c and LDH. The time was 0 weeks, 8 weeks, 10 weeks, 12 weeks, 16 weeks and 18 weeks, a total of 6 times; SOD and MDA content were tested in the 16th and 18th weeks.

[0051] 2.6.3 Aortic fatty plaque

[0052] Oil red staining of aorta: The experimental endpoint was 10 weeks after administration. After the animals were euthanized, the arteries from the aortic arch to the iliac artery were collected and stained with oil red to analyze the fatty plaques accumulated in the aorta.

[0053] 2.6.4 Cardiac weight-based outflow tract plaque status

[0054] At the end of the experiment, the heart and tibia were collected, and the heart was weighed and corrected with the tibia length to calculate the heart weight index, heart index = heart weight (g) / tibia length (cm) × 100%. The heart was fixed with 4% PFA and HE staining was used to evaluate the fat plaque in the outflow tract.

[0055] 2.7 Data Analysis

[0056] All data were entered into an Excel document and expressed as mean ± standard error. Significant differences were compared using one-way ANOVA analysis of variance. P values ​​less than 0.05 were considered to be significantly different, and P values ​​less than 0.01 were considered to be extremely significantly different.

[0057] 3. Results

[0058] 3.1 ApoE - / - Effect of changes in average body weight of mice

[0059] like Figure 1 As shown, HFD-fed ApoE - / -The average weight gain of mice was significantly higher than that of control mice. Starting from the 9th week, the weight of model, atorvastatin, and test drug AQ-Ca and DMS-ASD mice was significantly greater than that of the control (P < 0.05), and there was no significant difference between the groups (P > 0.05). The weight gain of mice in the test drug DMS group began to stabilize from the 10th to 11th week, and overlapped with the weight curve of the control group at the 14th week.

[0060] 3.2 ApoE - / - Effects of four blood lipids and plasma LDH content on mice

[0061] As shown in Table 1 and Figure 2 As shown in Figure 2, long-term HFD feeding led to increased dyslipidemia in mice. Compared with the control group, ApoE - / - After 8 weeks of HFD feeding, TC and LDC-c levels of mice increased significantly (P < 0.05), while TG and HDL-c levels did not change significantly (P > 0.05). Compared with the model group, atorvastatin significantly reduced TC and LDL-c levels. The test drug DMS showed a decrease in ApoE at 16-18 weeks. - / - The trend of TC and LDL-c in mice was not statistically significant (P>0.05). - / - There was no significant effect on TC and LDL-c in mice (P>0.05), but the HDL-c level in blood lipids was significantly increased at the 18th week (P<0.001).

[0062] At week 18, the plasma LDH of the model mice was significantly higher than that of the control group, and the plasma LDH of the atorvastatin and test drug groups, especially the DMS-ASD and DMS mice, was significantly lower than that of the model group (P < 0.05) (Table 1 and Figure 2 ).

[0063] Table 1. Drugs to be tested for ApoE - / - Effects of four blood lipids and LDH content on mice

[0064]

[0065]

[0066] 3.3 ApoE - / - Effects of plasma SOD and MDA on mice

[0067] After 8 weeks (week 16) and 10 weeks (week 18) of administration, the plasma SOD and MDA levels of mice were shown in Table 2 and Figure 3As shown in Table 2, there was no significant difference in SOD activity between the groups at week 16 and week 18 (P>0.05); the MDA content in the model group at week 18 was significantly lower than that in the control group (P<0.05), and there was no significant statistical difference between the other groups (P>0.05) (Table 2 and Figure 3 ).

[0068] Table 2. Drugs to be tested for ApoE - / - Effects of plasma SOD and MDA on mice

[0069]

[0070]

[0071] 3.4 ApoE - / - Effect of aortic plaque area ratio on mice

[0072] At the end of the 18th week of the experiment, the aorta of mice was stained with oil red and the atherosclerotic plaque area was quantitatively analyzed. The results are shown in Table 3. Figure 4 and Figure 5 As shown: The percentage of aortic plaques in the model group was significantly higher than that in the control group (12.71±1.88 vs 0.77±0.26, P<0.0001). Compared with the model group, the positive control atorvastatin significantly reduced the percentage of aortic plaques caused by HFD (P<0.01); the tested drugs AQ-Ca, DMS-ASD and DMS also showed the effect of reducing the percentage of aortic atherosclerotic plaques in mice, especially DMS-ASD and DMS compared with the model group, which were statistically significant (P<0.05).

[0073] Table 3. Drugs to be tested for ApoE - / - Effects of aortic plaque in mice

[0074]

[0075] 3.5 ApoE - / - Effect of heart weight index on mice

[0076] At the end of the experiment, the animals were euthanized and the heart was weighed and corrected by tibia length. The results are shown in Table 4 and Figure 6 As shown: Compared with the control group, there was no significant difference in the heart weight and heart weight index of mice in each group (P>0.05).

[0077] Table 4. Drugs to be tested for ApoE - / - Effect of heart weight index on mice

[0078]

[0079] IV. Conclusion

[0080] Administration of 300 mg / kg DMS-ASD and 20 mg / kg DMS for 10 consecutive weeks had multiple pharmacological effects of inhibiting the increase of mouse body weight, blood lipid TC, LDL-c and plasma LDH, and reducing the area of ​​aortic atherosclerotic plaques.

[0081] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. The use of myricetin diselenide for preparing a pharmaceutical composition, characterized in that: The pharmaceutical composition is used for preventing, alleviating and / or treating cardiovascular diseases.

2. The use according to claim 1, characterized in that The pharmaceutical composition is used for preventing and / or treating atherosclerosis.

3. The use according to claim 2, characterized in that The cardiovascular disease is selected from the group consisting of aortic atherosclerosis, coronary artery atherosclerosis, coronary atherosclerotic heart disease, carotid and cerebral artery atherosclerosis, renal artery atherosclerosis and limb artery atherosclerosis.

4. A pharmaceutical composition for preventing, alleviating and / or treating cardiovascular diseases, characterized in that: The active ingredient is myricetin diselenide.

5. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutical composition is used for preventing and / or treating atherosclerosis.

Images