Synthesis and biochemical testing of next generation peptide inhibitors of thrombosis
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The first interaction at a damaged blood vessel is between the multimeric glycoprotein von Willebrand Factor (vWF) and collagen that is now exposed. Binding of vWF enhances the rate of platelet binding at the site of injury resulting blood clot formation. The clotting of blood can be detrimental to the human body, for example, heart attacks and strokes which are leading causes of death. The purpose of this research was to synthesize cyclic peptides that can compete with vWF to bind to collagen with the goal of preventing the formation of a blood clot.Cyclic peptides HTNpep1_1BiPhe and HTNpep1_2BiPhe were synthesized by elongating the desired peptide chain, cyclizing from head-to-tail and thoroughly purifying. This research utilizes two unnatural amino acids to make the peptide less susceptible to degradation within the body and increase the likelihood of binding collagen. Both peptides’ stabilities were tested against specific proteases and were measured through multiple protease degradation assays. The peptides’ capability to compete with vWF when binding to collagen were tested using a new collagen-binding assessment of fluorescent-linked immunosorbent assay (FLISA) which was utilized in past research. The FLISA method tests a control to measure success of vWF binding collagen, a peptide from studied literature against vWF, and the two synthesized peptides (1BiPhe and 2BiPhe) against vWF. As anticipated, both peptides were synthesized and purified successfully. Protease degradation assay data provided evidence such that 1BiPhe does not degrade as much when compared to literature peptides and is very stable, meanwhile 2BiPhe is not as stable. The new FLISA method was performed only with 1BiPhe and was ultimately inconclusive with the exception of one trial, more trials will need to be further performed.
Department of Chemistry
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