The number of intracellular drug targets increased rapidly up to 71% of all targets, but are heavily underrepresented with FDA-approved drugs. Leaving 4,102 disease-associated drug targets untreated. Limited availability of drugs with intracellular targets is predominantly caused by absence of methods to quantify (un)bound subcellular drug concentrations. Hereinto, we developed a label-free technology relying on (1) rapid purification of organelles and the plasma membrane and (2) sampling of the cytosol and organelle matrix fractions. Our tag-based organelle and plasma membrane purification technology outperforms a previous approach more than 3-fold, with yields up to 59.1 ± 8.9% using transient expression. Next, organelle-specific fluorescent dyes were used to demonstrate specific and fast (~5 minutes) sampling of the cytosol and organelle lumen. Subsequently, cytosolic cyclosporin A and lysosomal imipramine concentrations were determined, which aligned with literature values. Quantifying a drug’s subcellular distribution will enhance the translation of intracellular disease targets to the patient.
Learning Objectives:
After this presentation, participants will understand the relevance of subcellular drug concentrations to enhance translation of intracellular drug targets into patients.
After this presentation, participants have acquired knowlegde about QTM's technology to quickly isolate organelles for subcellular drug quantification.
After this presentation, participants have acquired knowlegde about QTM's technology to sample the cytosol and organelle lumens for subcellular drug quantification.
