The Elucidation on mechanism of drug metabolism by a statistic and dynamics approaches.

上一财政年度的结果表明，三环类抗抑郁药可能是值得关注的目标，应在几种异构体上进行研究。特别是，CYP 2C 9和2C 19已被报道，这两种异构体在各自中代谢底物的相反性质。在这项工作中，我们专注于位置72和241上的两个相互残基，并分别制备了两种突变体;即，CYP 2C 9的K72 E和K241 E以及CYP 2C 19的E72 K和E241 K。对CYP 2C 9和2C 19上的这些突变体和野生型进行了评价，评价了它们对三种三环类抗抑郁药（阿米替林、丙咪嗪和多硫平）的结合亲和力和代谢活性。推测72位氨基酸可能对底物的识别特性有贡献，主要是对碱性药物。

The result in the last fiscal year suggested that tricyclic antidepressants could be interesting targets which should be investigated on several CYP isoforms. Especially, CYP2C9 and 2C19 had been reported that the two isoforms metabolized opposite properties of substrates in each. In this work, we focused on two reciprocal residues on positions 72 and 241, and two mutants were prepared, respectively; i.e., K72E and K241E for CYP2C9 and E72K and E241K for CYP2C19. These mutants and wild-types on CYP2C9 and 2C19 were evaluated on binding affinities and metabolic activities on three tricyclic antidepressants; amitriptyline, imipramine, and dothiepin. The estimated values implied that an amino acid located on position 72 could contribute to recognize property of substrate, mainly on basic drugs.