Application of ridecevir in preparation of medicine for preventing and treating atherosclerosis

Abstract

The invention relates to an application of ridecevir in preparation of a medicine for preventing and treating atherosclerosis. The application of the ridecevir in preparation of the medicine for preventing and treating the atherosclerosis comprises one or more of the following effects: (1) the ridecevir has an inhibiting effect on formation of atherosclerotic plaques; (2) the ridecevir plays a role in inhibiting the activation of vascular endothelium; (3) the ridecevir plays a role in reducing mRNA (messenger Ribonucleic Acid) and protein expression level of VCAM-1 and ICAM-1 mediated by ox-LDL (Lipoprotein Dehydrogenase Lipoprotein); (4) the ridecevir plays a role in reducing the immunofluorescence expression of ox-LDL mediated endothelial cells such as VCAM-1 and ICAM-1; and (5) reducing the adhesion of the ox-LDL mediated THP1 cell on the surface of the endothelial cell by the ridecevir. The invention provides new application of the ridecevir, namely application of the ridecevir or pharmaceutically acceptable salts, esters and hydrates of the ridecevir in preparation of medicines for preventing and treating atherosclerosis diseases. The invention provides an effective novel potential alternative drug for preventing and treating atherosclerosis, expands the adaptation of the ridegravir, and improves the application potential and market prospect of the ridegravir.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of remdesivir in the preparation of drugs for the prevention and treatment of atherosclerosis. Background Technology

[0002] Atherosclerosis is a common cardiovascular disease characterized by the formation of plaques in the arterial intima. These plaques are composed of lipids, fibrous tissue, necrotic material, and cells, and may lead to hardening of the blood vessel walls and narrowing of the lumen.

[0003] Atherosclerosis is a major cause of cardiovascular disease and stroke. Its pathological mechanism involves complex pathological processes such as endothelial dysfunction, which in turn mediates inflammatory responses.

[0004] The treatment of atherosclerosis is a comprehensive process aimed at slowing disease progression, relieving symptoms, preventing complications, and improving patients' quality of life. Existing treatment methods include: lifestyle modifications, drug therapy, surgical treatment, rehabilitation therapy, and psychotherapy.

[0005] Existing drug treatments include lipid-lowering drugs, antiplatelet drugs, antihypertensive drugs, and hypoglycemic drugs. Lipid-lowering drugs: Statins are commonly used lipid-lowering drugs that can lower low-density lipoprotein (LDL) levels, reduce plaque formation, and stabilize existing plaques. Antiplatelet drugs: Such as aspirin, can prevent platelet aggregation and reduce the risk of thrombosis. Antihypertensive drugs: For patients with hypertension, antihypertensive drugs are needed to control blood pressure and reduce damage to the blood vessel walls. Hypoglycemic drugs: For patients with diabetes, blood sugar levels need to be controlled to reduce damage to the blood vessel walls.

[0006] Remdesivir (L-Alanine, N-[(S)-hydroxyphenoxyphosphinyl]-,2-ethylbutyl ester,6-ester with 2-C-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2,5-anhydro-D-altrononitrile) is an adenosine nucleotide analog prodrug that works by inhibiting RNA-dependent RNA polymerase. It is primarily used to treat Ebola virus and is known for its broad-spectrum antiviral activity. Early data suggested that remdesivir showed efficacy in inhibiting SARS-CoV-2 replication and shortening hospital stays during the COVID-19 pandemic; however, later data from numerous clinical trials indicated that its anti-COVID-19 efficacy was not very significant. Nevertheless, research has found that remdesivir has potential therapeutic applications for non-viral inflammatory diseases.

[0007] Whether remdesivir can effectively treat atherosclerosis remains unclear. Therefore, this study focuses on exploring new applications of remdesivir in regulating vascular endothelial cell function and treating atherosclerosis, aiming to provide new treatment options for atherosclerotic cardiovascular diseases.

[0008] Therefore, we designed the application of remdesivir in the preparation of drugs for the prevention and treatment of atherosclerosis to solve the above problems. Summary of the Invention

[0009] To overcome the above shortcomings, this invention provides the application of remdesivir in the preparation of drugs for the prevention and treatment of atherosclerosis.

[0010] Furthermore, the remdesivir includes one or more of the following effects:

[0011] (1) The remdesivir described herein has an inhibitory effect on the formation of atherosclerotic plaques;

[0012] (2) The remdesivir described herein has an inhibitory effect on vascular endothelial activation;

[0013] (3) The remdesivir described herein reduces the mRNA and protein expression levels of ox-LDL-mediated VCAM-1 and ICAM-1;

[0014] (4) The remdesivir described herein reduces the immunofluorescence expression of VCAM-1 and ICAM-1 in ox-LDL-mediated endothelial cells;

[0015] (5) The remdesivir described herein reduces the adhesion of ox-LDL-mediated THP1 cells to the surface of endothelial cells.

[0016] Furthermore, a drug for preventing and treating atherosclerosis, the drug comprising remdesivir or its pharmaceutical salt, and a pharmaceutically acceptable carrier; the carrier comprising one or more of the following: diluent, buffer, suspension, emulsion, granule, encapsulation agent, excipient, filler, binder, spray, transdermal absorbent, humectant, disintegrant, absorption enhancer, surfactant, colorant, flavoring agent, or adsorbent.

[0017] Furthermore, the effective dose of remdesivir in the drug is as follows: if the standard adult weight is set at 60kg, then the effective dose for adults is 900mg every two days.

[0018] Furthermore, the dosage form is for oral or injectable administration, and the oral or injectable dosage form includes one of the following: powder, tablet, granule, capsule, oral liquid, emulsion, or suspension.

[0019] Furthermore, the remdesivir drug also contains other excipients.

[0020] The beneficial effects of this invention are:

[0021] The application of remdesivir in the preparation of drugs for the prevention and treatment of atherosclerosis, as described in this invention, includes the following effects:

[0022] (1) The remdesivir described herein has an inhibitory effect on the formation of atherosclerotic plaques;

[0023] (2) The remdesivir described herein has an inhibitory effect on vascular endothelial activation;

[0024] (3) The remdesivir described herein reduces the mRNA and protein expression levels of ox-LDL-mediated VCAM-1 and ICAM-1;

[0025] (4) The remdesivir described herein reduces the immunofluorescence expression of VCAM-1 and ICAM-1 in ox-LDL-mediated endothelial cells;

[0026] (5) The remdesivir described herein reduces the adhesion of ox-LDL-mediated THP1 cells to the surface of endothelial cells.

[0027] This invention provides a novel use for remdesivir, namely, the application of remdesivir or its pharmaceutically acceptable salts, esters, or hydrates in the preparation of drugs for the prevention and treatment of atherosclerotic diseases. This provides an effective new potential drug candidate for the prevention and treatment of atherosclerosis, expands the indications for remdesivir, and enhances its application potential and market prospects. Attached Figure Description

[0028] Figure 1A Oil Red staining image of the entire aorta of a mouse;

[0029] Figure 1B Statistical plot of Oil Red staining of the entire aorta of mice, *p<0.05, n=5;

[0030] Figure 1C Images of H&E staining and Oil Red O staining of the root of the aortic valve in mice, scale bar 100 μm;

[0031] Figure 1D Statistical graphs of H&E staining and Oil Red O staining of the aortic valve root in mice, *p<0.05, n=5. Mean inhibition rate = (1 - mean plaque area percentage in the remdesivir group / mean plaque area percentage in the control group) * 100%;

[0032] Figure 2A For RT-PCR detection of VCAM-1 and ICAM-1 mRNA levels in endothelial cells, *p<0.05, n=3;

[0033] Figure 2BFor the detection of VCAM-1 and ICAM-1 protein levels in endothelial cells by Western blot, *p<0.05, n=5;

[0034] Figure 2C For the detection of VCAM-1 and ICAM-1 protein levels in endothelial cells by Western blot, *p<0.05, n=5;

[0035] Figure 3A Cellular immunofluorescence: red represents VCAM-1 and ICAM-1, blue represents DAPI, scale bar is 50 μm, *p<0.05, n=3;

[0036] Figure 3B Cellular immunofluorescence: red represents VCAM-1 and ICAM-1, blue represents DAPI, scale bar is 50 μm, *p<0.05, n=3;

[0037] Figure 3C For the THP-1 adhesion assay, endothelial cells were treated with ox-LDL and remdesivir for 24 hours, THP-1 was labeled with a fluorescent dye, and then the cell adhesion assay was performed. The scale bar was 100 μm, *p<0.05, n=5.

[0038] Figure 3D For the THP-1 adhesion assay, endothelial cells were treated with ox-LDL and remdesivir for 24 hours, THP-1 was labeled with a fluorescent dye, and then the cell adhesion assay was performed. The scale bar was 100 μm, *p<0.05, n=5. Detailed Implementation

[0039] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the reagents used can be purchased commercially.

[0040] Example 1: Application Study of Preparation of Drugs for the Prevention and Treatment of Atherosclerosis

[0041] I. Experimental Materials

[0042] 1. Remdesivir: Purchased from Selleck, store in a cool, dry place at 4°C, protected from light.

[0043] 2. Experimental animals: APOE- / - mice (8-week-old males with a background of C57BL / 6, 18-20g), provided by Beijing Huafukang Biotechnology Co., Ltd.

[0044] 3. Human umbilical vein endothelial cells (HUVEC): isolated from the human umbilical vein.

[0045] 4. Solvents: Dimethyl sulfoxide (DMSO), corn oil.

[0046] II. Technical Methods

[0047] 1. Animal experiments

[0048] 1.1 Modeling and Grouping:

[0049] Eight-week-old male ApoE- / - mice were randomly divided into two groups of five (n=5) each: a control group (Ctrl group) and a remdesivir group (RDV group). Both groups of mice were fed a Western diet. The RDV group was administered the drug intraperitoneally at a dose of 15 mg / kg / qod, while the Ctrl group was administered an equal volume of DMSO (diluted with corn oil).

[0050] 1.2 Drug Preparation:

[0051] Remdesivir is first dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution with a concentration of 30 mg / ml, which can be stored at -20°C. Before administration, it is diluted with corn oil 10 times to prepare a 3 mg / ml formulation.

[0052] 1.3 Treatment and Observation:

[0053] After 12 weeks of feeding, the aorta of mice was dissected and stained with Oil Red O. Sections of the outflow tract of the mouse heart were stained with Oil Red O and H&E. Changes in atherosclerotic plaques in the aorta and outflow tract of the heart were observed in the two groups of mice.

[0054] 2. Cell experiments

[0055] 2.1 Modeling and Grouping:

[0056] Endothelial cells (HUVECs) were randomly divided into three groups and treated with DMSO, ox-LDL (100 μmol), and remdesivir (10 μmol) + ox-LDL (100 μmol) for 24 hours, respectively.

[0057] 2.2 Drug Preparation:

[0058] The expression of vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) in HUVECs was detected by RT-PCR, Western blot, and immunofluorescence (IF) to verify the effect of remdesivir on endothelial cell activation. The effect of remdesivir on endothelial cell adhesion was evaluated by monocyte adhesion assay (THP-1).

[0059] III. Experimental Results:

[0060] 1. Remdesivir inhibits the formation of atherosclerotic plaques.

[0061] Compared to the control group (Ctrl group), remdesivir treatment significantly inhibited the formation of atherosclerotic plaques. Oil Red O staining of the entire aorta of mice showed that 15 mg / kg / qod of remdesivir significantly inhibited the formation of atherosclerotic plaques in the entire aorta of mice, with an inhibition rate of 49.81%, which was statistically significant. Figures 1A-1B H&E staining and Oil Red O staining were performed on sections of mouse heart outflow tract. The results showed that 15 mg / kg / qod remdesivir significantly inhibited the formation of atherosclerotic plaques at the root of the aortic valve in mice, with inhibition rates of 21.44% and 27.60%, respectively, and the difference was statistically significant. Figures 1C-1D The effective dose of remdesivir in the drug is as follows: if the standard adult weight is set at 60kg, then the effective dose for adults is 900mg every 2 days.

[0062] 2. Remdesivir inhibits vascular endothelial activation.

[0063] Endothelial activation is considered a key step in the development of atherosclerosis. Activated endothelial cells induce the selective recruitment of monocytes to the vascular intima. Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), belonging to the immunoglobulin superfamily, mediate the adhesion between endothelial cells and leukocytes. In the critical stage of atherosclerosis, adhesion factors firmly attach monocytes to the endothelium. Therefore, this study aimed to verify whether remdesivir inhibited endothelial cell activation and thus regulated the inflammatory response by detecting the mRNA levels and protein expression of VCAM-1 and ICAM-1 in endothelial cells. The results showed that remdesivir significantly inhibited ox-LDL-mediated VCAM-1 and ICAM-1 mRNA and protein expression levels, with statistically significant differences. Figure 2A , Figure 2B and Figure 2CImmunofluorescence analysis showed that remdesivir also reduced ox-LDL-mediated immunofluorescence expression of VCAM-1 and ICAM-1 in endothelial cells, with statistically significant differences. Figures 3A-3B Furthermore, remdesivir significantly reduced ox-LDL-mediated adhesion of THP1 cells to the endothelial cell surface, with statistically significant differences. Figures 3C-3D The above results indicate that remdesivir inhibits the progression of atherosclerosis by suppressing ox-LDL-induced endothelial cell activation.

[0064] The conclusions that can be drawn from the experiment are:

[0065] Application of Remdesivir in the preparation of drugs for the prevention and treatment of atherosclerosis.

[0066] Remdesivir inhibits the formation of atherosclerotic plaques.

[0067] Remdesivir inhibits vascular endothelial activation.

[0068] Remdesivir reduces the mRNA and protein expression levels of ox-LDL-mediated VCAM-1 and ICAM-1.

[0069] The remdesivir described herein reduces the immunofluorescence expression of VCAM-1 and ICAM-1 in ox-LDL-mediated endothelial cells.

[0070] The remdesivir reduces ox-LDL-mediated adhesion of THP1 cells to the surface of endothelial cells.

[0071] The effective dose of remdesivir in the drug is as follows: if the standard adult weight is set at 60kg, then the effective dose for adults is 900mg every 2 days.

[0072] The remdesivir prepared is in the form of an oral or injectable dosage form, including powder, tablet, granule, capsule, oral liquid, emulsion or suspension.

[0073] The drug prepared from remdesivir also contains a pharmaceutically acceptable carrier, including diluents, buffers, suspensions, emulsions, granules, encapsulation agents, excipients, fillers, binders, sprays, transdermal absorbents, humectants, disintegrants, absorption enhancers, surfactants, colorants, flavoring agents, or adsorbents.

[0074] The remdesivir drug also contains other excipients.

[0075] In the description of this invention, it should be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0076] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art will understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0077] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of Remdesivir in the preparation of drugs for the prevention and treatment of atherosclerosis.

2. The application as described in claim 1, characterized in that: Remdesivir has one or more of the following effects: (1) The remdesivir described herein has an inhibitory effect on the formation of atherosclerotic plaques; (2) The remdesivir described herein has an inhibitory effect on vascular endothelial activation; (3) The remdesivir described herein reduces the mRNA and protein expression levels of ox-LDL-mediated VCAM-1 and ICAM-1; (4) The remdesivir described herein reduces the immunofluorescence expression of VCAM-1 and ICAM-1 in ox-LDL-mediated endothelial cells; (5) The remdesivir described herein reduces the adhesion of ox-LDL-mediated THP1 cells to the surface of endothelial cells.

3. A drug for preventing and treating atherosclerosis, characterized in that, The drug includes remdesivir or its pharmaceutical salts, and a pharmaceutically acceptable carrier; the carrier includes one or more of the following: diluent, buffer, suspension, emulsion, granule, encapsulation agent, excipient, filler, binder, spray, transdermal absorbent, humectant, disintegrant, absorption enhancer, surfactant, colorant, flavoring agent, or adsorbent.

4. According to claim 3, the effective dose of remdesivir in the drug is: if the standard adult weight is set at 60 kg, then the effective dose for adults is 900 mg every two days.

5. The drug for preventing and treating atherosclerosis according to claim 3 is an oral or injectable dosage form, wherein the oral or injectable dosage form includes one of the following: powder, tablet, granule, capsule, oral liquid, emulsion, or suspension.

6. The drug for preventing and treating atherosclerosis according to claim 3, characterized in that, The remdesivir drug also contains other excipients.

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