Elgaid, Zuhair; (2024) Exploration of the effectiveness and molecular mechanism of a new anti-EBV prototype drug. Doctoral thesis (Ph.D), UCL (University College London).

Text Zuhair Elgaid 19133020 PhD Thesis final version.pdf - Accepted Version Access restricted to UCL open access staff until 1 July 2025. Download (15MB)

Abstract

Multiple sclerosis (MS) is a neurodegenerative disease of the central nervous system, characterized by its chronic inflammatory and demyelinating pathology. Current MS treatments are safe and moderately effective, or more efficacious and potentially riskier. Epstein-Barr Virus (EBV) is the most defined risk factor and appears to be a prerequisite for MS onset. Epstein-Barr nuclear antigen 1 (EBNA-1) is an interesting drug target because it is the only viral protein expressed throughout the EBV cycle. Proteolysis targeting chimeras (PROTACs) consist of a protein of interest (POI) binder linked to a ubiquitin ligase recruitment ligand (UR). POI-PROTAC-UR complexes result in the polyubiquitination and proteasomal degradation of the target protein. The overall purpose of this thesis was to characterize an entirely new class of molecules that target EBNA-1. Molecular models were built to understand how current small molecule inhibitors bind EBNA-1. From this, a PROTAC version of EBNA-1 small molecule inhibitor VK-2019 was designed, and a fragment library was screened using computational docking. The EBNA-1 binding domain was then expressed and purified for biophysical assays. PROTACs and small molecule inhibitors produced an EBNA-1 response, but did not follow 1:1 Langmuir binding, likely causing a conformational change instead. 2 fragments, M-721 and M-296, were identified as potential starting points for a new EBNA-1 PROTAC. Cell based assays showed that EBNA-1 PROTAC JPC2 potentially reduced the efficiency of lymphoblastoid cell line generation and viability. JPC2 also affected EBNA-1 episome retention and degraded EBNA-1 via a PROTAC mechanism. Overall, this thesis details a first-generation EBNA-1 targeting PROTAC, with initial experiments showing promising results for future validation. A pipeline of tools and models have been developed to facilitate the design and development of future EBNA-1 PROTACs.

Download activity - last month

Export as CSVJSONXML

Download activity - last 12 months

Export as CSVXMLJSON

Downloads by country - last 12 months

Export as CSVXMLJSON

Archive Staff Only

View Item