PROJECT SUMMARY 14-3-3 proteins are at the crossroads of diverse cellular processes relevant to cancer, such as signal transduction, cell cycle progression and apoptosis. This family of hub proteins regulates these functions through an expansive network of protein-protein interactions (PPIs) that constitute the 14-3-3 “interactome.” Small molecule modulation of the 14-3-3 interactome is therefore of significant interest for cancer treatment, especially given the role of 14-3-3 proteins in resistance to standard of care drugs. Drugging the 14-3-3 signaling hub also enables multiple cancer-relevant processes to be perturbed in concert. Although modulation of 14-3-3 PPIs remains underdeveloped, the natural product cotylenin A presents a molecular platform to address this deficiency. Cotylenin A stabilizes 14-3-3/partner interactions by functioning as a rare “molecular glue.” This is believed to underlie cotylenin A’s notable anticancer activity and ability to sensitize cancer cells to existing treatments while sparing normal cells. Additionally, cotylenin A’s uncommon binding mode carries advantages of improved drug selectivity and lower affinities required for efficacy. However, cotylenin A has not been developed as a therapeutic due to loss of natural sources and lack of efficient and readily diversifiable syntheses. This proposal seeks to invigorate the development of cotylenin-based cancer therapies through chemical synthesis. Leveraging our group’s expertise in organic synthesis, we will develop an efficient and modular route to cotylenin A. Proof of concept for this synthetic strategy to access the cotylenin core has already been established, and will facilitate completion of the cotylenin A synthesis. The cotylenin core will enable immediate diversification at several positions to generate analogues based on existing crystallographic data. Modification of the synthesis at early stages will also allow structure-based diversification at additional sites, including one that provides a novel means to achieve specificity in PPI modulation. The anticancer activity of the analogues will be evaluated to elucidate structure-activity relationships. We will also conduct proteomic studies to identify the affected protein targets. This knowledge will guide iterative medicinal chemistry optimization of cotylenins. Research will be carried out at Scripps Research, an institution that excels in synthetic chemistry and chemical biology, and fosters strong collaboration across the two fields. I will receive training in complex molecule synthesis from Prof. Ryan Shenvi, a leader in the field, as well as training in chemical biology by performing biological and proteomic studies in collaboration with Prof. Chris Parker at Scripps Research. This work is complimentary to my doctoral training in the study of organometallic reaction mechanisms.