Now, as they report in Chem, Bortz, Bennett and Fasan leverage a cytochrome P450 site-selective C–H oxyfunctionalization as the key step to install functional groups as handles for directing chemical transformations of the scaffold, generating a library of complex small molecules, through a chemoenzymatic DOS (ceDOS) strategy.

The team used variants of fatty acid hydroxylase P450_BM3, which were previously engineered for the regio- and stereoselective transformations of the sesquiterpene lactone parthenolide (PTL), a model target substrate that has anti-cancer and anti-inflammatory activity, into 9(S)-hydroxy-PTL, 14-hydroxy-PTL, and 1,10-epoxy-PTL. The stereoselectively installed functional groups were treated as chemical handles enabling diversity-generating transformations of PTL via, but not limited to, acid-catalysed rearrangements and (macro)cyclizations, ring-opening or -closing metathesis reactions, oxidations and esterifications, 1,4-additions and Diels–Alder cycloadditions on installed dienophiles. Overall, this structural diversification effort led to a library of 51 compounds. Chemoinformatic tools confirmed the elevated degree of diversity and complexity of the library, which encompasses a chemical space larger than that achievable with more common late-stage C–H functionalization strategies. Additionally, proof-of-principle assays showed a range of biological activities against cancer cell lines.

现在，正如他们在 Chem 上报道的那样，Bortz、Bennett 和 Fasan 利用细胞色素 P450 位点选择性 C-H 氧官能化作为关键步骤，将官能团安装为指导支架化学转化的手柄，通过化学酶 DOS （ceDOS） 策略生成复杂小分子库。

该团队使用了脂肪酸羟化酶 P450_BM3 的变体，这些变体之前被设计为将倍半萜内酯小白菊内酯 （PTL）（一种具有抗癌和抗炎活性的模型靶标底物）的区域选择性和立体选择性转化为 9（S）-羟基-PTL、14-羟基-PTL 和 1,10-环氧-PTL。立体选择性安装的官能团被视为化学处理物，能够通过但不限于酸催化重排和（宏）环化、开环或闭合复分解反应、氧化和酯化、1,4-加成和已安装的亲二体上的 Diels-Alder 环加成反应来实现 PTL 的多样性产生转化。总体而言，这种结构多元化的努力导致了一个包含 51 种化合物的库。化学信息学工具证实了文库的多样性和复杂性程度的提高，它包含的化学空间比更常见的晚期 C-H 功能化策略所能实现的化学空间更大。此外，原理验证测定显示对癌细胞系的一系列生物活性。