A High Throughput Screening Approach Enabling Sustainable Discovery of Novel Lead Compounds from Natural Product Extracts

We propose the development of a high-throughput screening platform to facilitate the discovery of lead compounds from natural product extracts (NPEs). The proposed effort encompasses (a) utilization of NPEs from novel sources (cave organisms) with rich chemical diversity, (b) development of assays for specifically identified malaria targets, and (c) development of data management and analysis tools to automate the determination of optimal lead compounds from LC-MS data. In Phase I, selected cave microorganisms will be isolated and cultured to yield a chemical diverse `secondary metabolome'. Cost-effective, screen-friendly microbiological assays for critical P. falciparum pathogenicity (PfKASIII and hemozoin aggregation) will be developed. These validated assays will be used to quantify bioactivity of each culture condition in the extract library, along with simultaneous mass spectrometry, HPLC, and UV-Vis measurements. This wealth of data will be analyzed using a novel, Java-based Natural Extract Lead compound Identification (NELI) data management and analysis software. The primary innovative feature of the software is the identification of lead compounds using principal component analysis. In Phase II, rapid lead discovery of anti-malarial compounds by the NELI platform will be fully developed and demonstrated, and rendered ready for expansion to other disease types and natural product sources.