Codexis, Incorporated, 200 Penobscot Drive, Redwood City, CA 94063, USA.
Science. 2010 Jul 16;329(5989):305-9. doi: 10.1126/science.1188934. Epub 2010 Jun 17.
Pharmaceutical synthesis can benefit greatly from the selectivity gains associated with enzymatic catalysis. Here, we report an efficient biocatalytic process to replace a recently implemented rhodium-catalyzed asymmetric enamine hydrogenation for the large-scale manufacture of the antidiabetic compound sitagliptin. Starting from an enzyme that had the catalytic machinery to perform the desired chemistry but lacked any activity toward the prositagliptin ketone, we applied a substrate walking, modeling, and mutation approach to create a transaminase with marginal activity for the synthesis of the chiral amine; this variant was then further engineered via directed evolution for practical application in a manufacturing setting. The resultant biocatalysts showed broad applicability toward the synthesis of chiral amines that previously were accessible only via resolution. This work underscores the maturation of biocatalysis to enable efficient, economical, and environmentally benign processes for the manufacture of pharmaceuticals.
药物合成可以从酶催化相关的选择性增益中获益匪浅。在这里，我们报告了一种有效的生物催化过程，用以替代最近实施的铑催化不对称烯胺氢化反应，从而实现抗糖尿病化合物西他列汀的大规模生产。该过程从一种具有进行所需化学反应的催化机制但对前西他列汀酮没有任何活性的酶开始，我们采用底物行走、建模和突变方法，创建了一种对合成手性胺具有微小活性的转氨酶；然后，通过定向进化进一步对其进行工程化，以便在制造环境中实际应用。所得生物催化剂对合成手性胺具有广泛的适用性，而这些手性胺以前只能通过拆分来获得。这项工作强调了生物催化技术的成熟，使其能够实现高效、经济和环保的药物制造工艺。
