Use of indole acrylates for the preparation of medicaments for the treatment of hyperlipidemia and atherosclerotic cardiovascular diseases

Abstract

The present application belongs to the technical field of cardiovascular disease treatment, and particularly relates to application of indole propenoic acid in preparation of a medicine for treating hyperlipidemia and atherosclerotic cardiovascular disease. The present application aims to provide a new choice for treating hyperlipidemia and atherosclerotic cardiovascular disease. The technical scheme of the present application is the application of indole propenoic acid in preparation of a medicine for treating hyperlipidemia and cardiovascular disease. The present application discloses that IAA can effectively control body weight and regulate blood lipids without affecting food intake; can significantly reduce abdominal aortic plaque formation, lipid deposition of the heart aortic valve and AS lesions, and improve inflammation and endothelial function; provides a new idea and direction for treating hyperlipidemia and atherosclerotic cardiovascular disease, and has a broad application prospect.

Description

Technical Field

[0001] This invention belongs to the field of cardiovascular disease treatment technology, specifically relating to the application of indoleacrylic acid in the preparation of drugs for treating hyperlipidemia and atherosclerotic cardiovascular diseases. Background Technology

[0002] Atherosclerotic cardiovascular disease (ASCVD) is a collective term for a group of heart and blood vessel diseases, including coronary heart disease, myocardial infarction, and stroke. For many consecutive years, ASCVD has been the leading cause of morbidity and mortality worldwide. Between 2010 and 2020, deaths from cardiovascular diseases increased by 18.71%, accounting for 32% of global deaths, making it the leading cause of death from chronic diseases and a major global public health problem. Atherosclerosis is usually the underlying pathological cause of cardiovascular diseases such as coronary artery disease and myocardial infarction. Therefore, discovering effective treatments to control the development of atherosclerosis has become a key research direction for improving the cardiovascular health situation in China.

[0003] Recent studies have found a close relationship between gut microbiota and atherosclerotic cardiovascular disease. Gut microbiota has a significant impact on the host's physiological functions, such as metabolism and nutritional homeostasis, immune system maturation and activation, and even brain activity. Most of these effects are achieved through metabolites, either produced by the microbiota or derived from the transformation of molecules in the environment and the host. Tryptophan is one such metabolite resulting from the interaction between gut microbiota and the host. Tryptophan is an aromatic essential amino acid; among the 20 common amino acids, it has the largest molecular weight, but it is the least abundant amino acid in protein composition and cells. It is also a precursor for the biosynthesis of many microbial and host metabolites. Since animal cells do not produce tryptophan, humans rely on exogenous sources, primarily dietary intake. The World Health Organization recommends an intake of 4 mg / kg of tryptophan daily, and to date, no adverse effects from excessive tryptophan intake have been reported. Existing research has demonstrated the relationship between the gut bacteria-TMAO pathway and animal-derived foods and atherosclerotic cardiovascular disease. However, the role of indole-3-acrylic acid (IAA), a metabolite of chromoic acid, in atherosclerotic cardiovascular disease has not been reported. Summary of the Invention

[0004] The technical problem to be solved by this invention is to provide a new option for the treatment of hyperlipidemia and atherosclerotic cardiovascular disease.

[0005] The technical solution of the present invention is the application of indoleacrylic acid in the preparation of drugs for treating hyperlipidemia and / or cardiovascular diseases.

[0006] The technical solution of the present invention is the application of indoleacrylic acid in the preparation of products that improve hyperlipidemia and / or cardiovascular diseases.

[0007] Furthermore, the cardiovascular disease mentioned is atherosclerotic cardiovascular disease.

[0008] This invention also provides the application of indoleacrylic acid in the preparation of weight control products.

[0009] This invention also provides the application of indoleacrylic acid in the preparation of products that improve blood lipids.

[0010] This invention also provides the use of indoleacrylic acid in the preparation of products that reduce aortic plaque.

[0011] This invention also provides the use of indoleacrylic acid in the preparation of products that improve lipid deposition and / or atherosclerotic lesions of the aortic valve.

[0012] This invention also provides the use of indoleacrylic acid in the preparation of products that improve endothelial function.

[0013] This invention also provides the use of indoleacrylic acid in the preparation of products that reduce inflammation.

[0014] The present invention also provides a medicament for treating cardiovascular diseases, comprising indoleacrylic acid.

[0015] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0016] The present invention also provides products for improving hyperlipidemia and / or cardiovascular disease, including indoleacrylic acid.

[0017] Specifically, the cardiovascular disease mentioned is atherosclerotic cardiovascular disease.

[0018] The present invention also provides products for weight control, including indoleacrylic acid.

[0019] This invention also provides products for improving hyperlipidemia, including indoleacrylic acid.

[0020] The present invention also provides products for reducing aortic plaque, including indoleacrylic acid.

[0021] The present invention also provides products for improving lipid deposition and / or atherosclerotic lesions of the aortic valve, including indoleacrylic acid.

[0022] The present invention also provides products that improve inflammation and / or endothelial function, including indoleacrylic acid.

[0023] The present invention also provides products for reducing inflammation, including indoleacrylic acid.

[0024] Specifically, the aforementioned products are pharmaceuticals, food, or health supplements.

[0025] Beneficial effects of the present invention: The present invention constructs an ApoE for atherosclerosis - / - A mouse model was used to comprehensively detect atherosclerosis-related indicators (serum TC, TG, LDL-C, HDL-C, abdominal aortic Oil Red O staining, aortic valve Oil Red O staining, H&E, Masson staining, etc.). Results showed that IAA effectively controlled body weight and regulated blood lipids; it also significantly reduced abdominal aortic plaque formation, lipid deposition in the aortic valve, and atherosclerotic lesions, alleviated inflammation, and improved endothelial function. This demonstrates that IAA possesses pharmacological effects in improving atherosclerosis, providing a scientific basis for its development into potential drugs, foods, or health products for the prevention and treatment of hyperlipidemia and atherosclerotic cardiovascular diseases. This invention provides new ideas and directions for the treatment of hyperlipidemia and atherosclerotic cardiovascular diseases, with broad application prospects. Attached Figure Description

[0026] Figure 1 Statistical chart of weights in each group (n=12) Note: Compared with the control group: *P<0.05, **P<0.01, ***P<0.001.

[0027] Figure 2 Oil Red O staining images of the abdominal aorta in each group (n=3, ).

[0028] Figure 3 Oil Red O staining images of aortic valves in each group (n=3, ).

[0029] Figure 4 HE staining images of aortic valves in each group (n=3, ).

[0030] Figure 5 Masson staining images of aortic valves in each group (n=3, ).

[0031] Figure 6 Immunofluorescence staining images of ICAM1 in the aortic valve of each group (n=3, ).

[0032] Figure 7 Immunofluorescence staining images of VCAM1 in the aortic valve of each group (n=3, ).

[0033] Figure 8 , mRNA expression levels of inflammatory factor-related genes (n=12, ). Detailed Implementation

[0034] Experimental reagents: indoleacrylic acid, Western diet feed (No.: D200210, Deitz, USA).

[0035] Experimental animals: 7-8 week old male ApoE - / - Mice, weighing 18–22 g (purchased from Vital River Laboratory Animal Center, Beijing).

[0036] Example

[0037] All mice were randomly divided into three groups according to body weight using a random number table: a blank control group (ND), a model group (WD), a low-dose indoleacrylic acid group (IAA_L, 50 mg / kg), and a high-dose indoleacrylic acid group (IAA_H, 100 mg / kg), with 12 mice in each group. Male ApoE mice aged 7–8 weeks were used. - / - After one week of acclimatization, mice in the ND group were fed a normal diet, while the other groups were fed a Western diet. Simultaneously, mice in the IAA_L and IAA_H groups were given different doses of IAA (0.1 mL / 10 g), dissolved in 0.5% CMC-Na. The other groups received the same dose of 0.5% CMC-Na for 12 weeks. After 12 weeks of administration, all groups were fasted but allowed free access to water for 12 hours. Blood was collected via enucleation using isoflurane anesthesia. After blood collection, the mice's hearts were perfused with 30 mL of physiological saline. Following perfusion, the heart and abdominal aorta were removed sequentially.

[0038] Serum biochemical marker detection: for ApoE - / - After mice underwent the aforementioned grouping and corresponding drug interventions, serum levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol were determined by measuring four lipid parameters. Serum levels of nitric oxide and endothelin-1 were determined by measuring endothelial factor.

[0039] Histopathological examination: Heart tissue was collected and fixed in 4% paraformaldehyde for 24 h. It was then dehydrated using gradient concentrations of ethanol and xylene, embedded in paraffin, sectioned to a thickness of 4 μm, and stained with hematoxylin and eosin (HE) and Masson's stain according to the manufacturer's instructions. Frozen liver sections were prepared by dehydration in sucrose solution and then stained with Oil Red O solution for 30 min. The sections were then observed and photographed under a microscope.

[0040] Immunofluorescence staining: Paraffin blocks were sectioned at 5 μm intervals using a paraffin microtome. For paraffin sections, dewaxing was performed in xylene, followed by antigen extraction by boiling in citrate buffer (10 mM, pH 6.0) for 20 minutes, and then rehydration. After washing three times with phosphate-buffered saline, tissue sections were blocked for one hour in blocking solution (10% BSA and bovine serum in PBS), and then incubated overnight at 4°C in a humidified incubator with diluted primary antibodies (ICAM-1, Abcam, ab222736, IF: 1:50; VCAM-1, Abcam, ab134047, IF: 1:250). Sections were rinsed three times with PBS, incubated at 37°C for one hour with an appropriate Alexa Fluor 647 secondary antibody dilution (1:1000), rinsed three more times, and then mounted with DAPI-containing anti-fading mounting medium. All immunofluorescence micrographs were taken using an Olympus microscope.

[0041] Inflammatory factor gene expression: Aortic tissue was homogenized with trizol (Takara, Japan), extracted with chloroform, precipitated with isopropanol, washed with 75% ethanol, and dissolved in DEPC water. Concentrations were determined using an ultra-micro nucleic acid quantification system. PrimeScript was used. TM The RT reagent kit with gDNA Eraser (RR047A, Takara, Japan) reverse transcribed total RNA into cDNA. According to Takara's TB... Premix Ex Taq TM II (Tli RNaseH Plus) Operation Instructions: Real-time quantitative PCR (qRT-PCR) detection. PCR operating conditions: 95℃, 5 min, 40 cycles, each cycle 95℃, 10 s, 60℃, 30 s. The relative expression levels of Il-6, Il-1β, Tnf-α, and Mcp-1 were calculated using the 2^-ΔΔct method.

[0042] Data analysis and graphing were performed using GraphPad Prism 9.4.1 software. Two-tailed unpaired t-tests were used for comparisons between two groups, and one-way ANOVA and Tukey's multiple comparison test were used for comparisons among multiple groups. Data are expressed as mean ± standard error (x ± s), and P < 0.05 was considered statistically significant.

[0043] The experimental results are as follows:

[0044] The body weights of mice in each group are shown below. Figure 1Table 1 shows the food intake, indicating a significant decrease in the model group compared to the blank control group; however, there was no significant change in food intake between the high- and low-dose indoleacrylic acid groups compared to the model group. Based on the weight statistics, continued intake of both normal and Western-style diets can lead to ApoE. - / - The mice gradually gained weight, but their weight decreased significantly after indoleacrylic acid intervention.

[0045] Table 1. Food intake statistics for each group (n=12, )

[0046] Blank control group (ND) 19.775±0.143 Model Group (WD) 16.775±0.283* Low-dose indoleacrylic acid group (IAA_L) 16.715±0.315 High-dose indoleacrylic acid group (IAA_H) 16.667±0.268

[0047] Note: Compared with the blank group: * P<0.05.

[0048] The results of the four lipid profiles (Table 2) showed that, compared with the ND group, the WD group mice had significantly higher serum TC, TG, and LDL-C levels and significantly lower HDL-C levels. After intervention with different doses of IAA, the serum TC, TG, and LDL-C levels of the mice were significantly reduced, while the HDL-C level was significantly increased. This indicates that IAA can effectively improve the lipid levels of AS mice, reduce serum TC, TG, and LDL-C levels, and increase HDL-C levels.

[0049] Table 2. Statistical table of four blood lipid parameters for each group (n=12, )

[0050]

[0051]

[0052] Note: Compared with the blank group: *** P<0.001; Compared with the model group: △ P<0.05, △△ P<0.01, △△△ P<0.001.

[0053] Statistical results of Oil Red O staining of the abdominal aorta: Gross Oil Red O staining results of the abdominal aorta showed ( Figure 2 (Table 3) Compared with the ND group mice, the WD group mice had significantly increased abdominal aortic plaques, with the plaques mainly concentrated in the aortic arch. After intervention with different doses of IAA, the abdominal aortic plaques in the mice were significantly reduced. This indicates that IAA can significantly reduce the formation of abdominal aortic plaques.

[0054] Table 3. Statistical table of patch area for each group (n=3, )

[0055] Blank control group (ND) 2.348±0.628 Model Group (WD) 15.179±1.884*** Low-dose indoleacrylic acid group (IAA_L) <![CDATA[10.627±0.700 △ ]]> High-dose indoleacrylic acid group (IAA_H) <![CDATA[7.125±0.587 △△ ]]>

[0056] Note: Compared with the blank group:*** P<0.001; Compared with the model group: △ P<0.05, △△ P<0.01.

[0057] Oil Red O staining of the aortic valves of mice in each group showed ( Figure 3 (Table 4) Compared with the ND group, different degrees of lipid deposition were observed in the aortic valve of mice in the WD group and each drug administration group. Compared with the ND group, the lipid deposition area at the aortic valve of mice in the WD group was significantly increased. After different doses of IAA intervention, lipid deposition at the aortic valve of the heart was significantly reduced.

[0058] Table 4. Statistical table of patch area for each group (n=3, )

[0059] Blank control group (ND) 0.053±0.010 Model Group (WD) 0.496±0.032*** Low-dose indoleacrylic acid group (IAA_L) <![CDATA[0.380±0.019 △△ ]]> High-dose indoleacrylic acid group (IAA_H) <![CDATA[0.293±0.012 △△△ ]]>

[0060] Note: Compared with the blank group: *** P<0.001; Compared with the model group: △△ P<0.01, △△△ P<0.001.

[0061] like Figure 4 As shown in Table 5, compared with the ND group, different degrees of AS lesions, including atherosclerosis, fibrosis, and intimal thickening, were observed in the aortic valves of WD mice and mice treated with different doses. Compared with the ND group, WD mice had a large number of foam cells infiltrating the aortic valves, increased necrotic cores, and decreased fibrous cap thickness. After intervention with different doses of IAA, the foam cell infiltration in the aortic valves of mice decreased, the necrotic cores decreased, and the fibrous cap thickness increased, indicating that IAA has the effect of increasing plaque stability.

[0062] Table 5. Statistical table of plaque area, fibrous cap, and necrotic core for each group (n=3, )

[0063] Blank control group (ND) 0.0508±0.021 61.205±2.609 8.232±4.314 Model Group (WD) 0.530±0.045*** 23.952±6.124*** 42.896±2.997** Low-dose indoleacrylic acid group (IAA_L) <![CDATA[0.358±0.021 △△ ]]> 40.910±6.573 31.679±4.662 High-dose indoleacrylic acid group (IAA_H) <![CDATA[0.241±0.014 △△△ ]]> <![CDATA[46.502±4.068 △ ]]> <![CDATA[28.398±0.784 △ ]]>

[0064] Note: Compared with the blank group: *** P<0.001; Compared with the model group: △ P<0.05, △△ P<0.01, △△△ P<0.001.

[0065] like Figure 5 As shown in Table 6, compared with the ND group, the collagen fiber content at the aortic valve of the mouse heart was significantly increased in the WD group. After intervention with different doses of IAA, the collagen fiber content at the aortic valve of the mouse heart decreased.

[0066] Table 6. Statistical table of collagen fibers in each group (n=3, )

[0067] Blank control group (ND) 7.984±2.209 Model Group (WD) 28.398±5.158** Low-dose indoleacrylic acid group (IAA_L) 13.108±0.569△ High-dose indoleacrylic acid group (IAA_H) 11.398±3.357△

[0068] Note: Compared with the blank group: ** P<0.01; Compared with the model group: △ P<0.05.

[0069] As shown in Table 7, compared with the ND group, the serum NO level of mice in the WD group was significantly reduced and the ET-1 level was significantly increased. After intervention with different doses of IAA, the serum NO level of mice was significantly increased and the ET-1 level was significantly reduced, indicating that IAA can improve endothelial dysfunction in mice.

[0070] Table 7. Statistical table of NO and ET-1 for each group (n=3, )

[0071] Blank control group (ND) 679.812±28.794 124.110±2.098 Model Group (WD) <![CDATA[431.727±13.997 *** ]]> <![CDATA[169.247±1.968 *** ]]> Low-dose indoleacrylic acid group (IAA_L) <![CDATA[541.037±15.985 △△ ]]> <![CDATA[150.498±2.208 △△△ ]]> High-dose indoleacrylic acid group (IAA_H) <![CDATA[597.339±18.403 △△△ ]]> <![CDATA[138.821±2.293 △△△ ]]>

[0072] Note: Compared with the blank group: *** P<0.001; Compared with the model group: △△ P<0.01, △△△ P<0.001.

[0073] Immunofluorescence results showed ( Figure 6 , Figure 7 (See Table 8) Compared with the ND group, the expression levels of VCAM1 and ICAM1 genes in the abdominal aorta and aortic valve of mice in the WD group were significantly increased. After intervention with different doses of IAA, the expression levels of VCAM1 and ICAM1 genes were significantly reduced, indicating that IAA improves endothelial dysfunction in mice.

[0074] Table 8. Area statistics of ICAM1 and VCAM1 in each group (n=3, )

[0075] Blank control group (ND) 0.004±0.000 0.007±0.002 Model Group (WD) 0.065±0.010** 0.064±0.006*** Low-dose indoleacrylic acid group (IAA_L) 0.045±0.0008 <![CDATA[0.039±0.006 △△ ]]> High-dose indoleacrylic acid group (IAA_H) <![CDATA[0.023±0.011 △ ]]> <![CDATA[0.021±0.001 △△△ ]]>

[0076] Note: Compared with the blank group: ** P<0.01, *** P<0.001; Compared with the model group: △ P<0.05, △△ P<0.01, △△△ P<0.001.

[0077] The results of changes in the mRNA expression levels of inflammatory factor-related genes in the mouse abdominal aorta showed that ( Figure 8Compared with the ND group, the expression levels of Il-6, Il-1β, Tnf-α and Mcp-1 mRNA in the abdominal aorta of mice in the WD group were significantly increased. After intervention with different doses of IAA, the expression levels of Il-1β, Tnf-α and Mcp-1 mRNA in the abdominal aorta were significantly decreased, indicating that IAA can improve aortic inflammation in mice.

Claims

1. Application of indoleacrylic acid in the preparation of drugs for treating hyperlipidemia.

2. Application of indoleacrylic acid in the preparation of products that improve blood lipids.

Images