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Application of protein in preparation of medicine for preventing and treating atherosclerosis and complications

A technology for atherosclerosis and atherosclerosis, which is applied in the field of biomedicine, can solve the problems of limited research level and no drugs for oxLDL clearance, and achieve the effect of reducing the ratio level.

Pending Publication Date: 2020-03-17
SHANGHAI CLEAR FLUID BIOMEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

oxLDL is a key factor affecting the process of atherosclerosis, but limited by the current research level, most treatments targeting oxLDL are at the level of basic research
According to literature research, 3 oxLDL antibodies have applied for patents (patent numbers: US20040120893A1, US20070122419A1, CN101654481A), and 1 phase II clinical trial using oxLDL monoclonal antibody to evaluate the safety of atherosclerosis patients (ClinicalTrials.gov Clinical trial number: NCT01258907), but there is no effective drug for oxLDL clearance

Method used

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  • Application of protein in preparation of medicine for preventing and treating atherosclerosis and complications
  • Application of protein in preparation of medicine for preventing and treating atherosclerosis and complications
  • Application of protein in preparation of medicine for preventing and treating atherosclerosis and complications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Example 1. sDSS1 protein can interact with oxLDL or LDL.

[0115] 1.1 Experimental materials and methods

[0116] Experimental materials: sDSS1 protein, oxidized low-density lipoprotein (oxLDL) (Sulaibao, product number: H7980); low-density lipoprotein (LDL) (Solaibao, product number: H7960).

[0117] Experimental method: Add 2μg of oxLDL or LDL and 6μg of sDSS1 protein or 10μg of sDSS1 protein to 1.5ml under the conditions of 20mM sodium acetate / acetic acid buffer (pH4.5) or 20mM phosphate buffer (pH7.2) respectively Mix in the EP tube and incubate at 37°C for 12 hours. The incubation product was added with loading buffer, mixed evenly, and denatured at 100°C for 10 minutes to make a loading sample. The prepared samples were electrophoretically separated by polyacrylamide gel electrophoresis (SDS-PAGE). After separation, the SDS-PAGE gel was stained with Coomassie brilliant blue to display protein bands.

[0118] 1.2 Experimental results

[0119] In the acetate buf...

Embodiment 2

[0122] Example 2. sDSS1 protein can reduce the uptake of oxLDL by vascular endothelial cells.

[0123] 2.1 Experimental materials and methods

[0124]Experimental materials: sDSS1 protein, Dil-oxLDL (Thermo Fisher Scientific, catalog number: L34358), human umbilical vein endothelial cells (HUVEC) (PromoCell, catalog number: C-12200)

[0125] Experimental method: HUVEC cells were inoculated into 6-well plates according to the number of cells of 300,000 per well. After 24 hours of attachment, 1.5ml of 10μg / ml Dil-oxLDL was added, or 1.5ml of 10μg / ml Dil-oxLDL was added at the same time at a concentration of 2 μg / ml, 5 μg / ml, 10 μg / ml, 20 μg / ml of sDSS1 protein. After continuing to incubate in the incubator for 5 hours, the fluorescent signal in the cells was observed using a fluorescence microscope, and the cells were digested with trypsin to form single cells for flow cytometry to detect the fluorescence intensity.

[0126] 2.2 Experimental results

[0127] After adding Dil-...

Embodiment 3

[0128] Example 3. sDSS1 protein inhibits phagocytosis of oxLDL by macrophages.

[0129] 3.1 Experimental materials and methods

[0130] Experimental materials: sDSS1 protein, Dil-oxLDL, phorbol ester (PMA, Sigma-Aldrich Company, product number: P1585), human monocyte THP-1 (Cell Bank of Type Culture Collection Committee, Chinese Academy of Sciences, catalog number: SCSP- 567).

[0131] Experimental method: THP-1 cells were inoculated into 6-well plates according to the number of 250,000 cells per well, and the culture medium contained 100ng / mL PMA. After the cells were activated by PMA for 48 hours, they were replaced with fresh medium and cultured for four days to help the cells adhere to the wall. Discard the old medium, add 1.5ml 10μg / ml Dil-oxLDL, or add 1.5ml 10μg / ml Dil-oxLDL and add sDSS1 protein at the concentration of 2μg / ml, 5μg / ml, 10μg / ml, 20μg / ml. After continuing to incubate in the incubator for 5 hours, observe the fluorescent signal in the cells with a fluor...

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Abstract

The invention relates to an application of protein in preparation of medicines for preventing and treating atherosclerosis and cardiovascular and cerebrovascular diseases and peripheral vascular diseases related to atherosclerosis. According to the invention, sDSS1 protein is used for preparing the medicines for preventing and treating the diseases. The sDSS1 protein can effectively reduce the level of oxidized low-density lipoprotein in animal blood of an atherosclerosis model, inhibit the ingestion of vascular endothelial cells and macrophages on oxidized low-density lipoprotein, can increase the intake of oxidized low-density lipoprotein by liver cells and reduce the area of atherosclerotic plaques in model group animals, and inhibit the progress of atherosclerosis or the progress of cardiovascular and cerebrovascular diseases and peripheral vascular diseases caused by atherosclerosis, which has great clinical application value.

Description

technical field [0001] The content of the invention belongs to the field of biomedicine, and relates to the application of a protein in the preparation of drugs for preventing and treating atherosclerosis and atherosclerosis complications. Background technique [0002] Atherosclerosis refers to the dysfunction of arteries due to the accumulation of lipids in the arterial wall to form lipid streaks or atheromatous plaques. The decrease of arterial vasomotor function, lumen stenosis and even thrombus due to plaque formation will affect the blood supply of tissues and organs supplied by the artery, resulting in partial or total ischemia of tissues and organs. [0003] Epidemiological data, preclinical research and clinical trial data from 1955 to the present show that dyslipidemia characterized by elevated low density lipoprotein cholesterol (LDL-c) is a key factor in atherosclerotic cardiovascular disease. Disease (atherosclerotic cardiovascular disease, ASCVD) important risk...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P9/10
CPCA61K38/1709A61P9/10A61P25/28C07K14/4702A01K2227/105A01K2217/075A01K2267/0375A61K38/17C07K14/47C12N15/867
Inventor 张英豪王耀付晶鹏闫桂蕊
Owner SHANGHAI CLEAR FLUID BIOMEDICAL SCI CO LTD
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