Synthesis of Novel Chromenone Esters for Biochemical Assays

This project details the synthesis of a variety of novel chromenone esters to be used as enzymatic probes. A robust, initial library of 22 chromenone esters was successfully synthesized with differences in chain length, branching, and the incorporation of cyclic and heteroatomic moieties. These new chromenones were fully characterized by 1HNMR, 13CNMR, DEPT HRMS, 19FNMR, and HSQC. The utility of these compounds comes from the fact that they will fluoresce upon cleavage of the ester; thus, this property makes them ideal candidates to be used in the monitoring of enzymatic activity of esterases. Elucidating the enzyme-substrate structure activity relationship (SAR) uncovers specific structural motifs favored by the active site. As more information is acquired about the binding sites of these enzymes, more efficient chemical inhibitors can be created as treatments for infections like Mycobacterium tuberculosis (Mtb) and Staphylococcus aureus. At present, a standardization curve has been developed to normalize enzymatic data. As more experiments are performed, specific enzymatic characteristics, such as Km and kcat can be determined.