Project 1: Isolation Chemistry of Plants and U.S. Lichens and Biological Evaluation

PROJECT SUMMARY – PROJECT 1 Project 1 at The Ohio State University (OSU) is comprised of taxonomic authentication, isolation chemistry, biological testing, and compound synthesis components. Specific Aim 1 will involve receiving extracts of bioactive authenticated tropical plant materials from Project 2 (University of Illinois at Chicago, UIC) and from the NCI Natural Products Repository for phytochemical work-up in Aim 2, and extracting with solvents taxonomically authenticated U.S. coastal lichens and their cultured fungal associates (mycobionts), with these fungi identified from their 18S rDNA sequence. Specific Aim 2 will deal with (a) LC-MS dereplication studies on active leads following preliminary screening (in order to prioritize samples for activity-guided fractionation); (b) purification procedures using chromatographic methods (guided by both in-house bioassays and those elsewhere in the program project); (c) compound structure elucidation, and (d) scale-up isolation, respectively. The structures of bioactive compounds will be determined using modern spectroscopic methods, backed up by X-ray crystallography, where necessary. In Specific Aim 3, plant, lichen, and fungal extracts will be subjected to primary screening against a small panel of cancer cell lines at Core 1 (UIC). The Project 1 bioassays will involve four already installed rare cancer cell lines (BDCM leukemia, HPAC pancreatic, HT-1376 urinary bladder, and MDA-T32 thyroid) as well as target-based assays [viz., nuclear factor-B (NF-κB), and mitochondrial transmembrane potential (MTP), and annexin V-apoptosis testing, in addition to an assay to evaluate samples for pyroptosis effects]. These assays will be available also for the testing of compounds isolated in Projects 2 and 3 and synthesized in Core 2. A strong collaboration has been established to test promising compounds from Projects 1-3 at the Nationwide Children’s Hospital, Columbus, OH [NF2 (neurofibromatosis type 2)-associated vestibular schwannomas and meningiomas]. In addition, selected compounds will be submitted for formulation, stability, solubility, metabolism and pilot pharmacokinetic studies in Core 2 (OSU), and also tested in hollow fiber and xenograft in vivo assays in Core 1 (UIC). Assay refinement in Project 1 will be conducted in collaboration with the biostatistics component of Core A (OSU). Specific Aim 4 will use chemical synthesis to scale up the supplies of plant, lichen, and microbial-derived compounds and to assist in compound structure elucidation work using chemical interconversion methods.