Approaches Towards the Inhibition of Anti-Apoptotic Proteins
cam.restriction | thesis_access_open | |
cam.supervisor | Spring, David R | |
cam.supervisor.orcid | Spring, David R [0000-0001-7355-2824] | |
cam.thesis.funding | false | |
dc.contributor.author | iegre, jessica | |
dc.contributor.orcid | iegre, jessica [0000-0002-9074-653X] | |
dc.date.accessioned | 2019-05-02T14:38:23Z | |
dc.date.available | 2019-05-02T14:38:23Z | |
dc.date.issued | 2019-07-19 | |
dc.date.submitted | 2019-02-25 | |
dc.date.updated | 2019-05-01T16:06:51Z | |
dc.description.abstract | Anti-apoptotic proteins play a fundamental role in cell survival. Under physiological conditions, such proteins trigger apoptosis in defective or damaged cells only; under pathological conditions, however, they can be dysregulated allowing the cells to survive despite being harmful. Considering the importance of anti-apoptotic proteins in many physio-pathological roles, their specific inhibition is an attractive strategy to develop safe therapeutics. This thesis describes the inhibition of two classes of anti-apoptotic proteins: 1) Inhibition of the anti-apoptotic protein CK2 to develop novel anti-cancer molecules targeting pockets outside the well-conserved ATP-binding site: -Using a Fragment-Based-Drug-Discovery (FBDD) approach twelve small molecule inhibitors of CK2 were developed. The lead molecule, 3l, inhibited the catalytic activity of CK2α by binding in the cryptic αD pocket with a Kd of 4 μM. 3l stopped proliferation of colorectal cancer cells with a GI50 of 10 μM and presented improved drug-like properties and selectivity compared to previously reported inhibitors. Remarkably, 3l has the potential to be developed into a potent and selective anticancer drug. -Using a combination of rational-based approach and peptide stapling, twenty-two conformationally-constrained peptides were generated to target the protein-protein interaction (PPI) of CK2 and affect its function. The lead peptide, P7-F1C5, presented a novel, highly-functionalised constraint that allowed the molecule to become cell-permeable, exert its anti-proliferative activity in cancerous cells, and to become resistant to serum proteases. P7-F1C5 is the first macromolecule reported in the literature that binds to CK2α with sub-micromolar affinity (Kd 150 nM), and that can act as a chemical probe for targeting the PPI of CK2. 2) Inhibition of the anti-apoptotic Bcl-2 proteins to dissect their role in platelet activation and apoptosis. Bcl-2 proteins regulate cell lifespan; however, their role in non-nucleated platelets is not fully understood. The elucidation of these pathways in platelets is crucial to the development of selective anti-platelet therapeutics. To this end, this thesis describes the development and the first application of twenty-seven BH3-only peptides in human platelets highlighting how peptides can provide an alternative to conventional methodologies to study PPIs in platelets. The most promising peptide, P9-F5C5, engaged the anti-apoptotic protein Bcl-xL with 26 nM affinity and reviled a new role for the protein Bim in platelet activation. | |
dc.description.sponsorship | Trinity College | |
dc.identifier.doi | 10.17863/CAM.39376 | |
dc.identifier.uri | https://www.repository.cam.ac.uk/handle/1810/292226 | |
dc.language.iso | en | |
dc.publisher.college | Trinity College | |
dc.publisher.department | Chemistry | |
dc.publisher.institution | University of Cambridge | |
dc.rights | All rights reserved | |
dc.rights | All Rights Reserved | en |
dc.rights.uri | https://www.rioxx.net/licenses/all-rights-reserved/ | en |
dc.subject | stapled peptides | |
dc.subject | FBDD | |
dc.subject | Peptides | |
dc.title | Approaches Towards the Inhibition of Anti-Apoptotic Proteins | |
dc.type | Thesis | |
dc.type.qualificationlevel | Doctoral | |
dc.type.qualificationname | Doctor of Philosophy (PhD) | |
dc.type.qualificationtitle | PhD |
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