Thesis
Novel reactive fragment screening technologies
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2022
- Thesis identifier
- T17143
- Person Identifier (Local)
- 201862127
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Advances in scientists’ abilities to study the genome and connect genetics to phenotype and disease outcome are increasing the number of validated targets on an unprecedented scale. Therefore, it is crucial for scientists to be able to rapidly identify chemicals that can modulate the activity of such targets. However, one of the major bottlenecks in the identification of chemical binders is the slow and iterative design-make-test paradigm that is deeply ingrained in medicinal chemistry research. To address this, the contents of this thesis detail the development, optimisation and validation of a direct-to-biology high-throughput chemistry (D2B-HTC) approach towards the identification of novel binders. The D2B-HTC platform was combined with covalent or reactive fragment screening, which involves an intact-protein mass spectrometric readout. This readout acts as a measure of quality control, validating the chemical identity of any hits and avoiding false positive results that have hindered D2B efforts to-date. The developed D2B-HTC platform utilised the target-based PhotoAffinity Bit (PhABit) screening technology and involved the rapid plate-based synthesis of >1000 diazirine-based fragments and subsequent screening of the crude reaction products against protein targets. A key advantage of this technology is the rapid and iterative synthesis of second generation or alternative libraries to accelerate data acquisition. In this way, the technology enabled the investigation into the suitability of a novel 2-aryl-5-carboxytetrazole (ACT) photoreactive moiety for reactive fragment screening via carboxylate residue modification. Screening a 546-membered library of ACT-based fragments enabled the identification of novel binders for both BCL6 and KRASG12D. Finally, until now, high energy ultraviolet light (302 nm) has been used to activate alkyldiazirine and ACT functionalities, which is known to be highly damaging to proteins. Therefore, an alternative and novel energy transfer approach towards the activation of these functionalities was developed, utilising lower energy light (>365 nm) in combination with appropriate photosensitisers.
- Advisor / supervisor
- Bush, Jacob T.
- Tomkinson, Nicholas C. O.
- Resource Type
- Note
- Previously held under moratorium in Chemistry department (GSK) from 22/09/2022 until 22/10/2024. The confidentiality statement on each page of this thesis DOES NOT apply.
- DOI
- Funder
- Embargo Note
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File | 2024-10-29 | Embargo |