Electronic Theses and Dissertations
APPLICATION OF COMPUTATIONAL METHODS TO PRIORITIZE EXPERIMENTS FOCUSED ON BIOMOLECULAR DRUG TARGETS
Date of Award
Doctor of Philosophy
SK1 is an intracellular enzyme whose lipid kinase activity regulates the levels of bioactive lipids that are involved in normal homeostasis and disease. SK1 activity is implicated in various diseases including cancer, and inhibition of SK1 activity is a potential treatment for its implicated pathogenesis. However, without a high-throughput SK1 activity assay, screening and identification of SK1 inhibitors is a rigorous process that requires significant time and resources to yield potent lead compounds. In order to develop a high-throughput SK1 assay method, an optically active non-lipid SK1 substrate was designed for the purpose of monitoring activity in real-time. Evaluation of the substrate design and prioritization of candidate compounds for experimental screening as SK1 substrates/ inhibitors was undertaken via computational methods, specifically molecular docking into SK1 structures. Candidates prioritized by docking were synthesized and evaluated for substrate/ inhibitor activity against recombinant, purified SK1. While none of the candidates showed SK1 substrate activity, the most promising candidate showed weak inhibitor activity against SK1.GPR88 is a membrane receptor whose downstream signaling activity is involved in neurodevelopment, behavior, locomotion and multisensory processing. GPR88 is implicated in neuropsychiatric diseases such as bipolar disorder and schizophrenia, thus GPR88 is a potential target for treating its implicated diseases. Currently, there is no experimentally determined, three-dimensional structure of GPR88 available and its endogenous ligand is unknown. Due to these unknowns, the structural and molecular mechanisms through which GPR88 elicit its physiological and pathological implications are unclear, In order to de-orphan GPR88, we developed a 3D structural model and used the model to perform in silico screening of human metabolites as potential candidates for endogenous ligands. Candidate endogenous metabolitesviiprioritized through the in silico screening process were experimentally screened for agonist activity against GPR88. Among these candidates, 3-iodothyronamine proved to be a GPR88 agonist with nano-molar potency.In these two projects, we were able to utilize computational methods as a means to efficiently inform and prioritize experimental efforts. Using computational methods, we identified a novel weak SK1 inhibitor design that may be later optimized, and also successfully identified a GPR88 endogenous agonist.
Dissertation or thesis originally submitted to ProQuest
Raji, Babatunde Adebiyi, "APPLICATION OF COMPUTATIONAL METHODS TO PRIORITIZE EXPERIMENTS FOCUSED ON BIOMOLECULAR DRUG TARGETS" (2018). Electronic Theses and Dissertations. 2947.
Available for download on Saturday, May 25, 2024
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