天然化合物を用いたタンパク質相互作用制御物質のスクリーニング

抄録

Poor in targets for drug discovery is a significant problem to develop clinical drugs, today. Thus, novel targets and/or strategies for drug screening are greatly required. Protein-protein interaction is one of exciting strategies to overcome the difficulty in drug development. On the other hand, regulation of protein-protein interaction with small molecule compounds has been considered to be difficult because larger spaces take part in the protein-protein interaction than those were expected to be covered by small molecule compounds. To challenge this opposite concept, we developed screening method to estimate protein-protein interaction applicable for drug screening. There are several methods to observe protein-protein interaction, however, rapidity and easiness are important for high through-put screening. As the results, we chose alpha-screen and bimolecular fluorescence complementation (BiFC) assay as the first screening method (1-2). We have constructed cell-based and in vitro BiFC screening system, and carried out 10 screenings using about 130,000 samples in each screening. As the results of screening, although, the hit ratio is extremely low, we have found 4 active substances. Including low selective regulators for protein-protein interaction, the average molecular weight was about 800. These results suggested that regulators of protein-protein interaction should have relatively large molecular weight as expected. Thus, natural compounds are considered to be suitable sources for protein-protein interaction regulators. 1) T. K. Kerppola, Visualization of molecular interactions by fluorescence complementation. Nat Rev Mol Cell Biol., 7, 449-456 (2006). 2) M. L. MacDonald, J. Lamerdin, S. Owens, B. H Keon, G. K. Bilter, Z. Shang, Z. Huang, H. Yu, J. Dias, T. Minami, S. W. Michnick and J. K. Westwick. Identifying off-target effects and hidden phenotypes of drugs in human cells. Nat. Chem. Biol., 2, 329-337 (2006)