Chemoenzymatic Strategies for Molecular Discovery and Asymmetric Synthesis - Chemoenzymatic Strategies for Molecular Discovery and Asymmetric Synthesis Building upon the progress made under prior NIGMS support, this MIRA project aims to introduce and pursue innovative strategies and platforms for (a) driving the discovery of ‘natural product’-like macrocyclic peptides as potent and selective modulators of biomolecular interactions, and (b) developing biocatalytic strategies for asymmetric synthesis via ‘new-to-nature’ chemistries. Under R01 GM134076 support, we introduced efficient and highly versatile methodologies for generating libraries of genetically encoded macrocyclic peptides in bacterial cells or displayed on phage particles, and demonstrated the effectiveness of these 'natural product-like' macrocycles toward disrupting biomedically relevant protein-protein interactions with high potency and specificity. Building upon this progress, these methodologies and platforms will be further expanded and potentiated through (i) the development of novel methods for the programmable synthesis of topologically complex macrocyclic peptides, including polycyclic peptide scaffolds; (ii) their integration with chemical warheads for the development of covalent protein inhibitors and (iii) the implementation of a new system for target-driven discovery in complex intracellular environments. These systems will be validated and applied to the discovery of cyclopeptide modulators of challenging biomolecular targets, including membrane proteins. Ultimately, these technologies are expected to streamline both the development of chemical probes for interrogating cellular pathways and validating therapeutic targets, and the discovery of potential lead structures for drug development, thereby accelerating efforts in basic biomedical research, chemical biology, and drug discovery. Under R01GM098628 support, we have pioneered and demonstrated the use of engineered myoglobins and other hemoproteins as efficient and robust biocatalysts for stereoselective carbene transfer reactions. Building upon this work, this research will investigate and extend the scope of these metalloprotein catalysts to a range of new, asymmetric carbon-carbon and carbon-heteroatom bond forming transformations useful for the synthesis of optically active building blocks and complex organic scaffolds of direct value for medicinal chemistry and drug discovery. Computational design, mechanism-guided design, and high-throughput experimentation will be leveraged to expedite the discovery and optimization of hemoprotein-based carbene transferases with enhanced catalytic efficiency, expanded reactivity, and fine-tuned stereoselectivity. The synthetic value of these methodologies will be further highlighted through the stereoselective, chemoenzymatic synthesis of drug molecules and natural products. Overall, this research is expected to make available new methodologies for the efficient, selective, and sustainable synthesis and diversification of biologically active molecules.
