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Enzymatic activation of double-targeted 5′-O-l-valyl-decitabine prodrug by biphenyl hydrolase-like protein and its molecular design basis

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Abstract

A primary focus of this research was to explore the activation process and mechanism of decitabine (5-aza-2′-deoxycytidine, DAC) prodrug. Recently, it has been reported that biphenyl hydrolase-like protein (BPHL) can play an important role in the activation of some amino acid nucleoside prodrugs with a general preference for hydrophobic amino acids and 5′-esters. Therefore, we put forward a bold hypothesis that this novel enzyme may be primarily responsible for the activation process of DAC prodrug as well. 5′-O-l-valyl-decitabine (l-val-DAC) was synthesized before and can be transported across biological membranes by the oligopeptide transporter (PEPT1), granting it much greater utility in vivo. In this report, l-val-DAC was found to be a good substrate of BPHL protein (K m 0.59 mM; k cat/K m 553.69 mM−1 s−1). After intestinal absorption, l-val-DAC was rapidly and almost completely hydrolyzed to DAC and l-valine. The catalysis was mainly mediated by the BPHL hydrolase and resulted in the intestinal first-pass effect of l-val-DAC after oral administration in Sprague-Dawley rats with cannulated jugular and portal veins. The structural insights using computational molecular docking showed that BPHL had a unique binding mode for l-val-DAC. As a fundamental basis, the simulation was employed to explain the catalytic mechanism in molecular level. In conclusion, BPHL was at least one of the primary candidate enzymes for L-val-DAC prodrug activation. This promising double-targeted prodrug approach have more advantages than the traditional targeted designs due to its higher transport and more predictable activation, thereby leading to a favorable property for oral delivery.

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Acknowledgements

This work was financially supported from the National Nature Science Foundation of China (No. 81302722).

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Authors and Affiliations

  1. Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang, 110016, China

    Wenhui Tao, Dongyang Zhao, Mengchi Sun, Meng Li, Zhonggui He, Yinghua Sun & Jin Sun

  2. Key Laboratory of Structure-Based Drug Design and Discovery, Shenyang Pharmaceutical University, Ministry of Education, No. 103, Wenhua Road, Shenyang, 110016, China

    Xiangyu Zhang

  3. Municipal Key Laboratory of Biopharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang, 110016, China

    Jin Sun

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  1. Wenhui Tao
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  2. Dongyang Zhao
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Correspondence to Yinghua Sun or Jin Sun.

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All animal experiments in the present study were approved by the University Committee on Use and Care of Animals, Shenyang Pharmaceutical University.

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Tao, W., Zhao, D., Sun, M. et al. Enzymatic activation of double-targeted 5′-O-l-valyl-decitabine prodrug by biphenyl hydrolase-like protein and its molecular design basis. Drug Deliv. and Transl. Res. 7, 304–311 (2017). https://doi.org/10.1007/s13346-016-0356-1

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