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Hydroxylation Metabolisms of Crassicauline A in Rats Under Toxic Dose

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Abstract

Background and Objectives

Crassicauline A, a C19 diterpenoid alkaloid in Aconitum herbs, is an analgesic drug clinically used in China. The in vivo metabolism of crassicauline A is poorly understood, while potential bioactivation is anticipated via hydroxylation metabolism. This work, therefore, aimed to investigate the in vivo hydroxylation metabolism of crassicauline A in rats.

Methods

Using a de novo developed and validated UPLC–MS/MS method, excretion studies in rats were carried out to investigate the recoveries of crassicauline A and its hydroxylated metabolites in urine and feces. Mass fragmentation analysis was used to identify the detected hydroxylated metabolites. In vitro metabolism assay in liver S9 fraction was employed to preliminarily investigate the inter-species difference of hydroxylation metabolism between rats and human.

Results

At a toxic dose of 100 µg/kg, less than 10% and 5% of the administrated dose of crassicauline A were recovered in the urine and feces after single intravenous and oral administration, respectively. Trace of yunaconitine, a possible 3-hydroxylated metabolite of crassicauline A, was detected in urine samples, but not considered to be derived from the in vivo metabolism, because the recovered yunaconitine and crassicauline A was equivalent to their occurrences in the test article. Another hydroxylated metabolite was detected with much higher levels than yunaconitine. Based on chromatographic behaviors and fragmentation analysis, the hydroxylation site of this metabolite was tentatively identified at C-15 on the skeleton, which might have produced a toxic alkaloid known as deoxyjesaconitine. The in vivo observations were consistent with the preliminary in vitro results in liver S9 fraction, in which an inter-species difference was highlighted that rats demonstrated more hydroxylation than human did.

Conclusions

This work disclosed that crassicauline A is elimilated in rats predominantly by metabolism under toxic dosage and the hydroxylation probably at C-15 might be a potential bioactivation pathway in both rats and human.

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Acknowledgements

The authors thank Prof. Wang F. P. and Associate Prof. Chen D. L. from West China School of Pharmacy, Sichuan University for gifting 3-deoxyaconitine. This work was funded by the National Natural Science Foundation of China (No. 81302844).

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Authors and Affiliations

  1. Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu, China

    Xue Fan, Shan-Shan Yin, Liang Xu & Ke Lan

  2. Chengdu Health-Balance Pharmaceutical and Biomedical Tech. Co. Ltd., Chengdu, China

    Shan-Shan Yin, Xue-Jing Li & Ke Lan

  3. Chengdu BaiKang Institute of Pharmacology and Toxicology, Chengdu, China

    Xue Fan, Shan-Shan Yin, Kui Yang & Ke Lan

Authors
  1. Xue Fan
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  2. Shan-Shan Yin
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  4. Kui Yang
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  5. Liang Xu
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Corresponding author

Correspondence to Ke Lan.

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Funding

This work was funded by the National Natural Science Foundation of China (No. 81302844).

Conflict of interest

Xue-Jing Li is an employee Chengdu Health-Balance Pharmaceutical and Biomedical Tech. Co. Ltd. Kui Yang is an employee of Chengdu BaiKang Institute of Pharmacology and Toxicology. The authors report no additional conflicts of interest.

Ethical approval

The animal experiments were conducted in accordance with Guide for the Care and Use of Laboratory Animals. The protocol was approved by the Ethical Committee of West China School of Pharmacy, Sichuan University. This article does not contain any studies with human participants performed by any of the authors.

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Fan, X., Yin, SS., Li, XJ. et al. Hydroxylation Metabolisms of Crassicauline A in Rats Under Toxic Dose. Eur J Drug Metab Pharmacokinet 42, 857–869 (2017). https://doi.org/10.1007/s13318-017-0408-z

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