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Hydrolytic Metabolism of Withangulatin A Mediated by Serum Albumin Instead of Common Esterases in Plasma

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Abstract

Background and Objectives

The oral bioavailability of withangulatin A (WA) is low and may undergo first-pass metabolism because of the presence of two esters bonds. This study aimed to identify the hydrolysis behavior and mechanism of WA, thus enriching its structure-pharmacokinetic relationship.

Methods

The in vivo pharmacokinetic studies of WA in rats were first investigated, followed by in vitro assays including metabolic stability, phenotyping identification and metabolic kinetics assays. After screening out the responsible enzymes with higher catalytic capacity, molecular docking study was performed to demonstrate the interaction mode between WA and metabolic enzymes. Then, metabolites in human serum albumin (HSA) were identified by LC-TOF-MS/MS.

Results

In rats, the oral bioavailability of WA was only 2.83%. In vitro, WA was hydrolyzed in both rat and human plasma and could not be inhibited by selective esterase inhibitors. Physiologic concentration of HSA not recombinant human carboxylesterases (rhCES) could significantly hydrolyze WA, and it had a similar hydrolytic capacity with human plasma to WA. Furthermore, WA could stably bind to HSA by forming hydrogen bonds with Lys199 and Arg410, accompanied by the metabolic reaction of the lactone ring opening.

Conclusion

The study showed that WA underwent obvious hydrolysis in rat and human plasma, which implied a strong first-pass effect. Serum albumin rather than common esterases primarily contributed to the hydrolytic metabolism of WA in plasma.

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Acknowledgements

We thank Jia Liu from Pharmaceutical Animal Experimental Center of China Pharmaceutical University for guiding the animal experiments. We also express our appreciation to Xi Xu for his help in molecular docking study.

Funding

This work was supported by the National Natural Science Foundation of China (No. 81872746).

Author information

Author notes

  1. Yu Zhuang and Yuxiao Wang have contributed equally to this article.

Authors and Affiliations

  1. Department of Pharmacology, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, China

    Yu Zhuang

  2. Zhejiang Institute for Food and Drug Control, Hangzhou, China

    Yu Zhuang

  3. Department of Pharmacology, School of Pharmacy, China Pharmaceutical University, Nanjing, 210009, China

    Yuxiao Wang & Zhixia Qiu

  4. National Experimental Teaching Demonstration Center of Pharmacy, China Pharmaceutical University, Nanjing, China

    Ning Li

  5. Shimadzu (China) Co., LTD, Nanjing Branch, Nanjing, China

    Haitao Meng

  6. Jiangsu Key Laboratory of Drug Design and Optimization, Department of Medicinal Chemistry, China Pharmaceutical University, Nanjing, China

    Zhiyu Li

  7. Jiangsu Key Laboratory of Bioactive Natural Product Research and State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, 210009, China

    Jianguang Luo

Authors
  1. Yu Zhuang
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  2. Yuxiao Wang
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  3. Ning Li
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  4. Haitao Meng
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  6. Jianguang Luo
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  7. Zhixia Qiu
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Contributions

Experimental design: YZ, ZQ. Animal experiment: YZ, YW. LC-MS/MS analysis: YZ, NL, HM. Manuscript writing and revision: YZ, YW, ZL, JL, ZQ.

Corresponding authors

Correspondence to Jianguang Luo or Zhixia Qiu.

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Conflict of Interest

There are no conflicts of interest declared by all the authors.

Ethical Approval

The experiment was performed in accordance with the Internal Charter on the Humane Care and Use of Laboratory Animal and supported by Ethics Committee of China Pharmaceutical University (approval number: 2022-04-012).

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Supplementary Information

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Supplementary file1 (DOCX 171 KB)

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Zhuang, Y., Wang, Y., Li, N. et al. Hydrolytic Metabolism of Withangulatin A Mediated by Serum Albumin Instead of Common Esterases in Plasma. Eur J Drug Metab Pharmacokinet 48, 363–376 (2023). https://doi.org/10.1007/s13318-023-00834-8

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  • DOI: https://doi.org/10.1007/s13318-023-00834-8

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