Date of Award

2016

Document type

Thesis

Degree Name

MCh by research (Master of Surgery by research)

First Supervisor

Dr Marian Brennan

Second Supervisor

Dr Dermot Cox

Funder/Sponsor

This work was funded by a scholarship from the Kuwaiti Government. Molecular graphics and analyses were performed with the UCSF Chimera package. Chimera is developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIGMS P41-GM103311).

Keywords

Platelet Aggregation Inhibitors -- Analysis, Cardiovascular Diseases -- Treatment

Abstract

The binding of VWF to GpIb-α is a critical step for maintaining normal haemostasis. The interaction between VWF with GpIb initiates the adhesion and aggregation of platelets at an injury site in the blood vessel wall. This binding is specific to high shear conditions that are typically found in small arterioles where the shear rates vary between 500-5000 s-1 . However, excessive binding of GpIb-α and VWF under normal conditions can cause stroke or myocardial infarction (MI). The purpose behind blocking GpIb is to stop the initial adhesion and stimulation of platelets under high shear conditions. There have been several attempts to generate GpIb-α inhibitors, such as the monoclonal antibody H6B4, OS-1 peptide, and the snake venom anfibatide. However, only anfibatide has successfully reached the clinical trials. In this project, we aimed to design a novel GpIb inhibitor for the treatment of cardiovascular disease. We performed both high-throughput docking as well as pharmacophore based screening to identify lead compounds for development. We then performed platelet aggregation assays, high shear flow experiments to assess their function ex vivo, and ELISA assays to investigate the binding interaction between the tested compounds and GpIb. A pharmacophore was generated based on GpIb-α crystal structure and published mutagenic data. The cyclic peptide GKYFG from the pharmacophore screening inhibited the interaction between VWF and GpIb-α under high shear conditions at a concentration of 0.1 mM (P

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 License.

File Size

4.78 MB

Comments

A thesis submitted for the degree of Master of Science from the Royal College of Surgeons in Ireland in 2016.

Available for download on Saturday, June 09, 2018

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