Use of arglabin C in preparation of a medicament for regulating vascular smooth muscle cell function and preventing cardiovascular diseases

Abstract

The application discloses application of silvergrass alcohol C in preparation of a medicine for regulating vascular smooth muscle cell function and preventing and treating cardiovascular diseases, and belongs to the technical field of medicines.The compound silvergrass alcohol C can inhibit expression of adhesion molecules of a rat vascular smooth muscle cell (VSMC) induced by trimethylamine oxide (TMAO) and cell migration, and further inhibit adhesion of the VSMC to a bone marrow mononuclear macrophage induced by TMAO, so as to play a role in regulating vascular smooth muscle cell function; and the compound silvergrass alcohol C is used to prepare the medicine for regulating vascular smooth muscle cell function and preventing and treating cardiovascular diseases.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the application of silver thread grass alcohol C in the preparation of drugs for regulating vascular smooth muscle cell function and preventing cardiovascular diseases. Background Technology

[0002] Cardiovascular diseases (CVD) are a group of diseases affecting the heart and blood vessels, including atherosclerosis, coronary heart disease, cerebrovascular disease, peripheral artery disease, rheumatic heart disease, congenital heart disease, deep vein thrombosis and pulmonary embolism, as well as restenosis after angioplasty or stent implantation. Vascular smooth muscle cells (VSMCs) are the main cellular components of the vascular wall structure, playing a role in maintaining vascular wall integrity and vascular tone. Changes in their structure and function can affect various cardiovascular diseases such as atherosclerosis and hypertension. When the vascular wall is mechanically damaged, it triggers a series of inflammatory responses characterized by granulocytes, releasing various vasoactive substances such as coagulation factors, growth factors, and adhesion molecules, leading to excessive proliferation, migration, and adhesion of VSMCs to inflammatory cells, ultimately resulting in changes in vascular structure such as angiogenesis and intimal hyperplasia. In recent years, increasing evidence has shown that circulating microbial metabolites are associated with cardiovascular diseases. Trimethylamine N-oxide (TMAO), a metabolite produced by phosphatidylcholine, has been identified as a biomarker for predicting CVD risk and a therapeutic target for CVD. In particular, TMAO can upregulate the expression of pro-inflammatory adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1) and intracellular cell adhesion molecule-1 (ICAM-1), promoting monocyte adhesion to VSMCs. Therefore, regulating vascular smooth muscle function will provide a novel approach for the prevention and treatment of cardiovascular diseases.

[0003] In recent years, the number of new drugs approved by the U.S. Food and Drug Administration (FDA) shows that cardiovascular disease drugs account for less than 10% of all indications. Despite persistently high mortality and morbidity rates associated with cardiovascular disease, drug development has fallen short of expectations. To date, most drugs used clinically for cardiovascular disease have significant side effects, such as liver and kidney damage, rhabdomyolysis, induction of diabetes, accelerated aging, and heart failure, limiting their clinical application. For life-threatening cardiovascular disease patients, treatment primarily involves surgery, including angioplasty, stent placement, plaque resection, bypass surgery, and drug-eluting stents such as paclitaxel. These treatments are expensive and have high restenosis rates. Therefore, finding new drugs and treatment strategies that specifically regulate vascular smooth muscle dysfunction caused by external factors to treat cardiovascular disease remains a pressing public health issue that needs to be addressed. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a pharmaceutical application of a compound called *Symplocos lucida* alcohol C, which upregulates GSTpi protein function and is used to inhibit VSMC proliferation and migration and monocyte adhesion for the prevention and treatment of cardiovascular diseases. *Symplocos lucida* alcohol C is a natural compound derived from plants of the *Chloranthus* genus. While its anti-inflammatory effects have been reported in the literature, its activity against cardiovascular-related diseases has not yet been reported.

[0005] This invention provides the application of compound silver-thread alcohol C in the preparation of drugs for the prevention and treatment of diseases related to vascular intimal hyperplasia.

[0006] This invention also provides the application of compound silver-thread alcohol C in the preparation of drugs for the prevention and treatment of cardiovascular-related diseases.

[0007] In one embodiment of the present invention, the vascular intimal hyperplasia-related diseases include atherosclerosis, restenosis after angioplasty, and restenosis after stent implantation.

[0008] In one embodiment of the present invention, during the application process, the compound *Symplocos lucida* C exhibits an inhibitory effect on the proliferation and migration of VSMCs. This demonstrates that the compound plays a crucial role in the development and progression of atherosclerosis, as well as in restenosis after angioplasty and stent implantation. Therefore, the compound *Symplocos lucida* C of the present invention can be used to prepare anti-atherosclerotic drugs and drugs for treating and alleviating restenosis after angioplasty or stent implantation.

[0009] This invention provides the application of silver thread grass alcohol C in the preparation of drugs for regulating vascular smooth muscle cell function and preventing cardiovascular diseases.

[0010] In one embodiment of the present invention, the structural formula of the compound silver thread grass alcohol C is shown below:

[0011]

[0012] [9,21-Dihydroxy-5-(hydroxymethyl)-23-(1-methoxy-1-oxopropane-2-ethylene)-13,20-dimethyl-4,22-dioxo-3-oxaoctadecylbutenoic acid-5,16(24)-dien-9-yl]methyl(E)-2-methyl-2-ethyl ester.

[0013] In one embodiment of the present invention, the compound silver-thread alcohol C is a natural compound extracted from plants of the genus Chloranthus.

[0014] In one embodiment of the present invention, the compound silver thread grass alcohol C is used to prepare a drug that upregulates glutathione S-transferase Pi (GSTpi).

[0015] In one embodiment of the present invention, the compound silver thread herbicide C is used to prepare a drug that inhibits the migration of vascular smooth muscle cells (VSMCs).

[0016] In one embodiment of the present invention, the compound silver thread grass alcohol C is used in the preparation of a drug for inhibiting the adhesion of vascular smooth muscle cells.

[0017] In one embodiment of the present invention, the compound silver thread alcohol C is used to prepare a drug for treating cardiovascular diseases.

[0018] The present invention also provides the use of the above-mentioned silver thread herb alcohol C in the preparation of a medicament for treating atherosclerosis.

[0019] The present invention also provides the above-mentioned silver thread herb alcohol C in the preparation of a drug for treating and relieving restenosis after angioplasty or stent implantation.

[0020] The natural compound silver-derived herbal extract C in this invention has the use of mediating the upregulation of glutathione S-transferase Pi (GSTpi) protein function, and particularly relates to its ability to regulate the function of vascular smooth muscle cells (VSMCs), and its application in the preparation of drugs for regulating vascular smooth muscle cell function to prevent and treat cardiovascular diseases.

[0021] This invention reveals that at the cellular level, strychnine C significantly inhibits TMAO-induced expression of adhesion molecules, cell migration, and its effect on monocyte / macrophage adhesion in rat VSMCs. Western blot results show that strychnine C significantly inhibits the expression of TMAO-induced rat VSMC adhesion molecules VCAM-1 and ICAM-1; scratch assays show that strychnine C significantly inhibits TMAO-induced rat VSMC migration; and cell adhesion assays show that strychnine C significantly inhibits TMAO-induced rat VSMC adhesion to BMDM. Therefore, the compound strychnine C of this invention has good application prospects and can be used to prepare drugs for regulating VSMC cell function and treating cardiovascular diseases.

[0022] In one embodiment of the present invention, the dosage form of the drug includes decoction, pill, powder, paste, elixir, wine, syrup, extract, lozenge, stick, suppository, medicated paste, and roasted paste.

[0023] In one embodiment of the present invention, the dosage form of the drug includes tablets, granules, blister packs, oral liquids, capsules, pills, mixtures, tinctures, aerosols, films, injections, and injectables.

[0024] In one embodiment of the present invention, the drug further contains medically acceptable excipients, including binders, fillers, disintegrants, lubricants, antioxidants, flavoring agents, solubilizers, emulsifiers, solubilizers, osmotic pressure regulators, and colorants.

[0025] In one embodiment of the present invention, the drug further comprises a pharmaceutical carrier.

[0026] In one embodiment of the present invention, the pharmaceutical carrier includes microcapsules, microspheres, nanoparticles, and liposomes.

[0027] The beneficial effects of this invention are as follows:

[0028] (1) The compound silver thread grass alcohol C has a strong inhibitory effect on the proliferation and migration of VSMCs;

[0029] (2) The compound silver thread grass alcohol C can upregulate the expression of VSMC adhesion molecules and can be used to prepare drugs for the prevention and treatment of cardiovascular diseases;

[0030] (3) The compound silver thread grass alcohol C can play a better role in preventing and treating diseases related to vascular intimal hyperplasia (such as atherosclerosis and cardiovascular diseases such as restenosis after angioplasty) by upregulating GSTpi protein.

[0031] (4) The compound silver thread herb alcohol C exhibits the effect of inhibiting the proliferation and migration of VSMCs, proving that it plays a key role in the occurrence and development of atherosclerosis, as well as restenosis after angioplasty and restenosis after stent implantation. Therefore, the compound silver thread herb alcohol C of the present invention can be used to prepare drugs for anti-atherosclerosis, and can be used to prepare drugs for treating and relieving restenosis after angioplasty or restenosis after stent implantation. Attached Figure Description

[0032] Figure 1 The effect of the compound strychnine C on the viability of vascular smooth muscle cells was investigated. (A) Cell viability of vascular smooth muscle cells after 24 h of stimulation with different concentrations of strychnine C (0, 2.5, 5, 10, 20, 40, 80 μM); (B) Cell viability of vascular smooth muscle cells after 48 h of stimulation with different concentrations of strychnine C (0, 2.5, 5, 10, 20, 40, 80 μM). The results showed that strychnine C concentrations less than or equal to 20 μM and stimulation time of 24 h had no significant effect on the viability of vascular smooth muscle cells. *p < 0.05, **p < 0.01, ***p < 0.001.

[0033] Figure 2 This study investigated the effect of the compound *Symplocos lucida* C on the expression of TMAO-induced cell adhesion molecules in vascular smooth muscle cells. Results showed that *Symplocos lucida* C (0, 2.5, 5, 10, 20 μM) significantly inhibited the expression of basal and TMAO-induced cell adhesion molecules VCAM-1 and ICAM-1 in vascular smooth muscle cells in a concentration-dependent manner. Compared with the Control group, **p < 0.01, ***p < 0.001; compared with the TMAO-treated group, ## p < 0.01, ### p < 0.001.

[0034] Figure 3 This study investigated the effect of compound strychnine C on TMAO-induced vascular smooth muscle cell migration. Results showed that strychnine C (20 μM) significantly inhibited TMAO-induced vascular smooth muscle cell migration at both 24 h and 48 h. ***p < 0.001.

[0035] Figure 4 This study investigated the effect of compound strychnine C on TMAO-induced adhesion between vascular smooth muscle cells and bone marrow mononuclear macrophages. Results showed that strychnine C (20 μM) significantly inhibited TMAO-induced adhesion between vascular smooth muscle cells and bone marrow mononuclear macrophages. ***p < 0.01.

[0036] Figure 5This study investigated the effect of compound strychnine C on vascular intimal hyperplasia in a TMAO-enhanced mouse carotid artery injury model. Results showed that strychnine C (75 mg / mL) significantly inhibited vascular intimal hyperplasia following TMAO-enhanced carotid artery injury in mice. ***p < 0.001. Detailed Implementation

[0037] The present invention will be further described below with reference to the embodiments. Due to space limitations, the description of the experimental process cannot be very detailed. All parts not described in detail in the experiment are conventional operations well known to those skilled in the art. However, the embodiments are by no means any limitation on the present invention.

[0038] Unless otherwise specified, the methods used in the following examples are conventional methods, and the reagents required in the following implementations are all commercially available unless otherwise specified.

[0039] The data analysis in the examples all used Graphpad Prism 8.0 statistical analysis software for one-way ANOVA multiple group comparisons.

[0040] Example 1: Effect of compound silver thread grass alcohol C on the viability of rat VSMC cells

[0041] Rat vascular smooth muscle cells (VSMCs) were treated with compound strychnine C (0, 2.5, 5, 10, 20, 40, 80 μM) for 24 h and 48 h, and cell viability was measured using the MTT assay. The method is briefly described as follows: Rat VSMCs were cultured with the above concentrations of compound strychnine C for 24 h and 48 h, and then incubated with 0.5 mg / mL MTT at 37 °C for 4 h. After dissolving in DMSO, the absorbance was measured at 490 nm.

[0042] The results are as follows Figure 1 As shown, treatment with silver thread grass alcohol C (5-40 μM) for 24 h had no effect on VSMC cell viability. Data are presented as mean ± standard deviation, *p<0.05, **p<0.01, ***p<0.001.

[0043] Example 2: Effect of compound silver-striped oxalis C on TMAO-induced expression of adhesion molecules in vascular smooth muscle cells

[0044] Rat VSMCs were pre-incubated with the compound silver-striped oxalis C (0, 2.5, 5, 10, 20 μM) for 2 h, followed by TMAO stimulation for 24 h. The expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) proteins was detected by Western blot. The Western blot method was briefly described as follows: total protein was extracted from VSMC cells and quantified. 30 μg of the total protein was separated by SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was blocked with 5% skim milk, and the antibody was incubated overnight at 4°C. After incubation with secondary antibody for 2 h, chemiluminescence reagent was used for detection.

[0045] The results are as follows Figure 2 As shown, silver thread herbicide C (0, 2.5, 5, 10, 20 μM) can dose-dependently inhibit the expression of cell adhesion molecules in rat VSMCs induced by TMAO; data are mean ± standard deviation. Compared with the control group, **p < 0.01, ***p < 0.001; compared with the TMAO treatment group, ## p < 0.01, ### p < 0.001.

[0046] Example 3: Effect of compound silver thread herbicide C on TMAO-induced migration ability of vascular smooth muscle cells

[0047] VSMCs were pretreated with the compound styraxone C (20 μM) for 2 h and stimulated with TMAO (500 μM) for 24 h. The migration ability of rat VSMCs was determined by the cell scratch assay. The method is briefly as follows: monolayer VSMCs were scratched with the tip of a 200 μL micropipette. After 24 h and 48 h, the cells were photographed under a microscope (Zeiss Axio Vert A1) and the cell migration distance was analyzed.

[0048] The results are as follows Figure 3 As shown, silver thread herbicide C (20 μM) significantly inhibited TMAO-induced migration of rat VSMCs (***p<0.001). Scale bar: 50 μm. Data are presented as mean ± standard deviation, ***p<0.001.

[0049] Example 4: Effect of compound silver thread grass alcohol C on TMAO-induced adhesion between vascular smooth muscle cells and bone marrow mononuclear macrophages

[0050] VSMCs were pretreated with the compound styraxol C (20 μM) for 2 h, stimulated with TMAO (500 μM) for 24 h, and then BMDM cells pre-incubated with BCECF-AM dye were added to the pretreated VSMC wells and incubated for 1 h. The number of BMDM cells adhering to the VSMCs was photographed and analyzed using a fluorescence microscope (Zeiss Axio Vert A1).

[0051] The results are as follows Figure 4 As shown, silver thread herbicide C (20 μM) significantly inhibited TMAO-induced adhesion between rat VSMCs and bone marrow mononuclear macrophages (***p<0.001). Scale bar: 20 μm. Data are presented as mean ± standard deviation, ***p<0.001.

[0052] Example 5: Effect of compound silver thread herbicide C on vascular intimal hyperplasia in a TMAO-enhanced mouse carotid artery injury model

[0053] Mice were anesthetized with sodium pentobarbital. The neck skin was disinfected with povidone-iodine, and a midline incision was made in the neck. The external carotid artery was ligated, the common carotid artery was clamped, and the proximal end of the external carotid artery was cut. A metal guidewire was inserted and rotated 3-5 times to establish a carotid artery injury model. One day post-surgery, mice were administered TMAO (86 μM) via intraperitoneal injection for 4 consecutive weeks. Two hours before each TMAO administration in week 4, mice were treated with silver thread grass alcohol C (75 mg / mL) by gavage. One week after treatment, vascular tissue was collected for paraffin sectioning and H&E staining.

[0054] Control group: sham surgery group, i.e., opening a cavity but not damaging blood vessels.

[0055] Model group: Damage control group treated with TMAO;

[0056] Treatment group (Model + silver thread herbicide C): Silver thread herbicide treatment group after vascular damage and TMAO treatment.

[0057] The results are as follows Figure 5 As shown, silver thread herbicide C significantly inhibited TMAO-enhanced carotid artery injury-induced intimal hyperplasia in mice (***p<0.001). Scale bar: 100 μm. Data are presented as mean ± standard deviation, ***p<0.001.

Claims

1. The application of compound silver thread herb alcohol C in the preparation of drugs for the prevention and treatment of diseases related to vascular intimal hyperplasia, wherein the diseases related to vascular intimal hyperplasia are atherosclerosis, restenosis after angioplasty, and restenosis after stent implantation.

2. The application according to claim 1, characterized in that, The dosage forms of the medicine include decoctions, pills, powders, ointments, medicated wines, syrups, extracts, tablets, and koji.

3. The application according to claim 1, characterized in that, The dosage forms of the drugs include tablets, granules, blister packs, oral liquids, capsules, pills, mixtures, tinctures, and injections.

4. The application according to claim 1, characterized in that, The drug also contains medically acceptable excipients, including binders, fillers, disintegrants, lubricants, antioxidants, flavoring agents, solubilizers, emulsifiers, solubilizers, osmotic pressure regulators, and colorants.

5. The application according to claim 1, characterized in that, The drug also contains a pharmaceutical carrier; the pharmaceutical carrier includes microcapsules, microspheres, nanoparticles and liposomes.

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