Asymmetric reduction of 2-chloro-3-oxo-ester into enantiomerically high pure diltiazem precursor by a Candida ketoreductase

Abstract Methyl (2R,3S)-3-(4-methoxyphenyl)glycidate [(2R,3S)-MPGM] is an advanced chiral synthon for the synthesis of the cardiovascular drug diltiazem. It can be easily accessed by cyclizing the reduction products of methyl 2-chloro-3-(4-methoxyphenyl)-3-oxo-propanoate (1a). Herein, we report an identified carbonyl reductase (CpKR) from Candida parapsilosis that displayed an excellent stereoselectivity toward the keto substituent at the C3-position of the 2-chloro-3-oxo-ester 1a. The engineered Escherichia coli cells harboring CpKR gene were directly applied for the asymmetric reduction of keto ester 1a with a space-time yield of 46 g L−1 d−1, which represents the highest productivity in bio-reduction of 1a reported so far. The isolated chiral alcohol products were then applied to the chemical synthesis of (2R,3S)-MPGM in 99% ee and a total yield of 76% in the two-step chemo-enzymatic reactions, which far exceeded the maximum theoretical yield (50%) of the existing industrial process based on a lipase-catalyzed resolution of racemic MPGM. This work provides a promising eco-friendly and cost-effective route toward industrial synthesis of pharmaceutically relevant diltiazem.