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Rapalogues as hCES2A Inhibitors: In Vitro and In Silico Investigations

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Abstract

Background and Objective

Rapamycin and its semi-synthetic analogues (rapalogues) are frequently used in combination with other prescribed medications in clinical settings. Although the inhibitory effects of rapalogues on cytochrome P450 enzymes (CYPs) have been well examined, the inhibition potentials of rapalogues on human esterases have not been investigated. Herein, the inhibition potentials and inhibitory mechanisms of six marketed rapalogues on human esterases are investigated.

Methods

The inhibitory effects of six marketed rapalogues (rapamycin, zotarolimus, temsirolimus, everolimus, pimecrolimus and tacrolimus) on three major esterases, including human carboxylesterases 1 (hCES1A), human carboxylesterases 2 (hCES2A) and butyrylcholinesterase (BuChE), were assayed using isozyme-specific substrates. Inhibition kinetic analyses and docking simulations were performed to investigate the inhibitory mechanisms of the rapalogues with strong hCES2A inhibition potency.

Results

Zotarolimus and pimecrolimus displayed strong inhibition of human hCES2A but these agents did not inhibit hCES1A or BuChE. Further investigation demonstrated that zotarolimus could strongly inhibit intracellular hCES2A in living HepG2 cells, with an estimated IC50 value of 4.09 µM. Inhibition kinetic analyses revealed that zotarolimus inhibited hCES2A-catalyzed fluorescein diacetate hydrolysis in a mixed manner, with the Ki value of 1.61 µM. Docking simulations showed that zotarolimus could tightly bind on hCES2A at two district ligand-binding sites, consistent with its mixed inhibition mode.

Conclusion

Our findings demonstrate that several marketed rapalogues are potent and specific hCES2A inhibitors, and these agents can serve as leading compounds for the development of more efficacious hCES2A inhibitors to modulate the pharmacokinetic profiles and toxicity of hCES2A-substrate drugs (such as the anticancer agent irinotecan).

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Authors and Affiliations

  1. Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, People’s Republic of China

    Cheng-Cheng Shi

  2. Department of Pharmacology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, Henan, People’s Republic of China

    Cheng-Cheng Shi, Shi-Tong Chen & Li-Rong Zhang

  3. Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, People’s Republic of China

    Yun-Qing Song, Rong-Jing He, Xiao-Qing Guan, Li-Lin Song, Meng-Ru Sun & Guang-Bo Ge

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Correspondence to Cheng-Cheng Shi, Guang-Bo Ge or Li-Rong Zhang.

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Funding

This work was partially funded by the National Natural Science Foundation of China (81922070 & 81773815).

Conflict of interest

Cheng-Cheng Shi, Yun-Qing Song, Rong-Jing He, Xiao-Qing Guan, Li-Lin Song, Shi-Tong Chen, Meng-Ru Sun, Guang-Bo Ge and Li-Rong Zhang declare that there is no conflict of interest.

Authors’ contributions

LZ and GG conceived the study and designed the experimental process. CS, YS, MS, LS and XG performed the experiments; CS, SC, RH, GG and LZ carried out data and results analysis; GG, LZ and CS wrote the manuscript and interpretation of data.

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Shi, CC., Song, YQ., He, RJ. et al. Rapalogues as hCES2A Inhibitors: In Vitro and In Silico Investigations. Eur J Drug Metab Pharmacokinet 46, 129–139 (2021). https://doi.org/10.1007/s13318-020-00659-9

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