Modulating Drug Transporters Dynamics to Improve Therapeutic Efficacy

forthcoming

"The therapeutic effect of a drug is largely determined by how effectively it reaches its site of action. Drug distribution in the human body is defined by an intricate network of transporters, offering potential for dynamically controlling where a drug spends most of its time. DynamITE aims to provide the tools to manipulate this ""highway network"" of drug transport at will. DynamITE will exploit the common mechanism shared by transporters, that is the alternating-access. Every transporter must dynamically switch between different states to enable directional transport of its substrate through the biological membrane. At the molecular level, this involves important local and global changes in the protein's three-dimensional structure. The foundational methodology of DynamITE is a structural mass spectrometry approach that reports on such changes: hydrogen-deuterium exchange coupled to mass spectrometry (HDX-MS). I have demonstrated that HDX-MS can fingerprint the effects of drugs on the dynamic switching between states. DynamITE builds on this knowledge to identify and design modulators of transporter dynamics. A modulator that prevents the conformational changes underlying transport will block drug transport, while a modulator that increases the rate of conformational transitions will accelerate transport. DynamITE’s approach will be applied to selected drug transporters with proven critical roles in antibiotic resistance, drug toxicity, and drug transport into the brain. In parallel, DynamITE will advance the HDX-MS technique to make it applicable to living cells. Such a development will enable the observation of protein dynamics in cellulo, with a molecular level of resolution—something currently inaccessible. This methodological breakthrough be used to monitor drug transporters dynamics while in action within the cell. The toolbox developed within DynamITE will be applicable to any transporter for which targeted modulation of transport activity is desired. "