Related Experiment Video
Updated: Jun 16, 2026
Avidity-based Extracellular Interaction Screening (AVEXIS) for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions
Published on: March 5, 2012
Identification of two antagonists of the scavenger receptor CD36 using a high-throughput screening model
1Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Insights
Researchers developed a high-throughput screening assay to find new drugs targeting CD36, a protein linked to atherosclerosis. Two novel compounds were identified that inhibit CD36 activity and reduce lipid accumulation, offering potential for anti-atherosclerotic therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- CD36 is a scavenger receptor involved in lipoprotein uptake and linked to atherosclerosis.
- Developing antagonists for CD36 is a therapeutic strategy for atherosclerosis.
Purpose of the Study:
- To develop a high-throughput screening (HTS) assay for identifying CD36 antagonists.
- To discover novel small molecules that inhibit CD36-mediated lipoprotein uptake.
Main Methods:
- Expressed human CD36 in Spodoptera frugiperda (Sf9) cells using a baculovirus system.
- Utilized a HTS assay measuring the uptake of labeled acetylated low-density lipoprotein (DiI-AcLDL).
- Validated identified compounds in Chinese hamster ovary (CHO) cells and RAW 264.7 macrophage foam cell assays.
Main Results:
- Established IC(50) values for known CD36 ligands (Ox-LDL, Ac-LDL, HDL).
- Identified two novel inhibitory compounds (2016481B and 2038751B) with IC(50) values of 17.4 and 23.7 microM.
- Confirmed inhibition of DiI-AcLDL uptake and lipid accumulation in relevant cell models.
Conclusions:
- The developed HTS assay is effective for identifying CD36 antagonists.
- The novel compounds show potential as starting points for developing anti-atherosclerotic agents.
- Targeting CD36 offers a promising therapeutic avenue for atherosclerosis treatment.
Abstract:
CD36, a class B scavenger receptor, is an integral membrane protein that mediates the endocytosis of modified lipoproteins. The functions of CD36 are complex and have been associated with atherosclerosis. In the current study, we developed a high-throughput screening (HTS) assay to identify small molecule antagonists by expressing human CD36 using a Bac-to-Bac baculovirus expression system in Spodoptera frugiperda (Sf9) cells. Uptake of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate-labeled acetylated low-density lipoprotein (DiI-AcLDL) revealed that the IC(50) values for the CD36 ligands oxidatively modified LDL (Ox-LDL), Ac-LDL, and high-density lipoprotein (HDL) were 0.039, 0.019, and 0.010 microg/ml, respectively. Using the HTS assay, two novel compounds, 2016481B and 2038751B, were found to inhibit DiI-AcLDL uptake in insect cells and exhibited IC(50) values of 17.4 and 23.7 microM, respectively. These two novel compounds also inhibited DiI-AcLDL uptake in cultured Chinese hamster ovary (CHO) cells permanently expressing human CD36. Furthermore, these two compounds inhibited lipid accumulation in RAW 264.7 murine macrophage cells in foam cell assays. This HTS assay represents a potential method for identifying more effective macrophage scavenger receptor antagonists, which may serve as starting points for the development of novel anti-atherosclerotic agents.
