Enzyme-Mediated Two-Step Regio- and Stereoselective Synthesis of Potential Rapid-Acting Antidepressant (2S,6S)-Hydroxynorketamine
Enzyme-Mediated Two-Step Regio- and Stereoselective Synthesis of Potential Rapid-Acting Antidepressant (2S,6S)-Hydroxynorketamine
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DOI：
10.1021/acscatal.9b05384
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发表时间：
2020-04-03
期刊：
影响因子：
12.9
通讯作者：
Urlacher, Vlada B.
中科院分区：
文献类型：
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作者：
Bokel, Ansgar;Ruehlmann, Ansgar;Urlacher, Vlada B.
最近，麻醉剂(S)-氯胺酮已被批准为一种速效长效抗抑郁药。其代谢物(2S,6S)-羟诺氯胺酮已被发现具有类似的抗抑郁作用，但副作用较小，这使得该化合物成为一个有趣的合成目标。利用Thermobifida fusca YX细胞色素P450 154E1的第一球诱变，构建了一个三突变体，使(S)-氯胺酮有效地产生(2S,6S)-羟诺氯胺酮。该工程P450单加氧酶催化连续的氧化n -去甲基化和高度区域和立体选择性的c6 -羟基化反应，以85%的产物选择性直接生成所需的产物。将这种选择性单加氧酶整合到大肠杆菌全细胞生物催化剂中，可以以半制备规模生产(2S,6S)-羟基诺氯胺酮。对产物进行了纯化，并对其结构进行了核磁共振鉴定。
Recently, the anesthetic (S)-ketamine has been approved as a rapid-acting and long-lasting antidepressant. Its metabolite, (2S,6S)-hydroxynorketamine, has been found to have a similar antidepressant effect but with less undesirable side effects, which make this compound an interesting target for synthesis. Using the first-sphere mutagenesis of the cytochrome P450 154E1 from Thermobifida fusca YX, we constructed a triple mutant that enables the effective production of (2S,6S)-hydroxynorketamine from (S)-ketamine. This engineered P450 monooxygenase catalyzes the consecutive oxidative N-demethylation and highly regio- and stereoselective C6-hydroxylation reactions leading directly to the desired product with 85% product selectivity. The integration of this selective monooxygenase into an Escherichia coli whole-cell biocatalyst allowed the production of (2S,6S)-hydroxynorketamine at a semipreparative scale. The metabolite was purified and its structure was confirmed by NMR spectroscopy.