Split-pool synthesis of 1,3-dioxanes leading to arrayed stock solutions ofsingle compounds sufficient for multiple phenotypic and protein-binding assays

Diversity-oriented organic synthesis offers the promise of advancing chemic al genetics, where small molecules are used to explore biology. While the s plit-pool synthetic method is theoretically the most effective approach for the production of large collections of small molecules, it has not been wi dely adopted due to numerous technical and analytical hurdles. We have deve loped a split-pool synthesis leading to an array of stock solutions of sing le 1,3-dioxanes. The quantities of compounds are sufficient for hundreds of phenotypic and protein-binding assays. The average concentration of these stock solutions derived from a single synthesis bead was determined to be 5 .4 mM in 5 muL of DMSO. A mass spectrometric strategy to identify the struc ture of molecules from a split-pool synthesis was shown to be highly accura te. Individual members of the 1,3-dioxane library have activity in a variet y of phenotypic and protein-binding assays. The procedure developed in this study allows many assays to be performed with compounds derived from indiv idual synthesis beads. The synthetic compounds identified in these assays s hould serve as useful probes of cellular and organismal processes.