Ultra-small scale assay of enzyme inhibition for drug discovery

Piccolo Biosystems, Inc., Seattle WA

Investigators

Abstract

Project Summary Lipid pathways are recognized as critical to cell function and survival and are crucial in disease states. Targeting the lipid-processing enzymes in these pathways has become increasingly important and promising in a wide range of diseases, including cancers, autoimmune, and neurological diseases. For example, there are currently >40 pharmacologic drugs targeting the sphingolipid pathway alone in clinical trials or in clinical use for oncology and autoimmune diseases. Despite their importance, massive gaps exist in our understanding of these pathways due to technical limitations in measuring the biochemical activity involved in lipid signaling, particularly in a high-throughout, highly sensitive, and cost-effective approach. To meet this need, Piccolo Biosystems Inc., an early stage biotechnology company, will collaborate with scientists at the University of Washington to develop and validate an innovative assay for measuring and targeting an important drug target sphingosine kinase (SphK) in cancer cells. Piccolo will leverage our patent-pending technology - the Picoliter-scale Thin Layer Chromatography (pTLC) platform to enable the assays of lipid-modifying enzymes in an automated fashion. In this Phase I SBIR, this collaboration will validate the sample transferring workflow to ensure the repeatable spoQing of pL- volume samples, optimize lipid separation conditions, and validate the in vitro assay conditions with a focus on quantitative strategies to benchmark performance. While this proposal specifically focuses on SphK for cancers, the validated workflow will enable high-content/high-throughput assays in a rugged and reproducible manner.