Optimizing peptides with activity as next generation anti-thrombosis agents

Abstract

Both heart attacks and stroke continue to be the leading causes of death in America and both can be attributed to pathogenic blood clots. As arteries become damaged by atherosclerotic plaques collagen becomes exposed; this collagen binds to the blood protein von Willebrand Factor (vWF). This vWF attracts platelets to the damaged area and initiates cross-linking, which can then lead to thrombosis. The main goal of this project is to develop a cyclic peptide that can target and inhibit and the initiation of the clot formation. Building off of previous work, this project consists of substituting unnatural amino acids into our expanded rings to improve the potency and stability. The ability of the peptides to inhibit the vWF-collagen interaction will be tested, along with their stability of their structure against degrading factors. In order to test these capabilities two different assays will be used; Enzyme-Linked Immunosorbent Assay (ELISA) and a protease degradation assay. ELISA quantifies the cyclic peptides’ abilities to displace vWF from binding collagen. The protease assay quantifies the peptides’ stability in a cellular-type environment. Along with these assays, this project includes synthesizing peptides and purification techniques.