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Isolation and identification of human metabolites from a novel anti-tumor candidate drug 5-chlorogenic acid injection by HPLC-HRMS/MSn and HPLC-SPE-NMR

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Abstract

A novel anti-tumor candidate drug, 5-chlorogenic acid (5-CQA) injection, was used for the treatment of malignant glioma in clinical trial (phase I) in China. The isolation and identification of the metabolites of 5-CQA injection in humans were investigated in the present study. Urine and feces samples obtained after intramuscular administration of 5-CQA injection to healthy adults have been analyzed by high-performance liquid chromatography coupled with high-resolution mass and multiple-stage mass spectrometry (HPLC-HRMS/MSn). No metabolite was detected in human feces; however, in human urine, a total of six metabolites were identified including isomerized 5-CQA (P1 and P2), hydrolyzed 5-CQA (M1and M2), and methylated 5-CQA (M3 and M4). Among them, M3 and M4 were the main metabolites and target analytes for human mass balance study. Additionally, the structure of M3 and M4 was characterized by high-performance liquid chromatography-solid phase extraction-nuclear magnetic resonance (HPLC-SPE-NMR), and the results demonstrated that the methoxy group of M3 and M4 was exclusively attributed to C-3′ and C-4′, respectively. Due to the unavailability of commercial reference, the pure products of M3 and M4 were synthesized by 5-CQA methylation and followed by isolation and purification. Moreover, the potential activity of M3 and M4 on malignant glioma was predicted using a reverse molecular docking analysis on eight malignant glioma-related pathways. The results showed that M3 and M4 had various interactions against malignant glioma-related targets. Our study provides an insight into the metabolism of 5-CQA injection in humans and supports the clinical human mass balance study.

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Abbreviations

CQA:

Chlorogenic acid

HMBC:

Heteronuclear multiple bond correlation

HPLC-HRMS/MSn :

High-resolution mass and multiple-stage mass spectrometry

HPLC-SPE-NMR:

High-performance liquid chromatography-solid phase extraction-nuclear magnetic resonance

HSQC:

Heteronuclear single-quantum correlation

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Acknowledgements

The Ministry of Science and Technology of the People’s Republic of China (2016ZX09101017) is acknowledged.

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Author notes

  1. Tiankun Ren and Yanan Wang contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Peking Union Medical College & Chinese Academy of Medical Sciences, No. 2 Nanwei Street, Xicheng District, Beijing, 100050, China

    Tiankun Ren, Yanan Wang, Caihong Wang, Jiandong Jiang & Jinlan Zhang

  2. Jiuzhang Biochemical Engineering Science and Technology Development Co., Ltd., Chengdu, Sichuan, 610041, China

    Mengtian Zhang & Wang Huang

  3. Department of Glioma, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, China

    Wenbin Li

Authors
  1. Tiankun Ren
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  2. Yanan Wang
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  3. Caihong Wang
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  4. Mengtian Zhang
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  5. Wang Huang
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  8. Jinlan Zhang
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Corresponding authors

Correspondence to Jiandong Jiang, Wenbin Li or Jinlan Zhang.

Ethics declarations

Informed consent was obtained from all individual participants who provided urine and feces samples. Ethics approval for this study has been obtained from the Ethics Committee, Beijing Shijitan Hospital, China.

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The authors declare that they have no conflicts of interest.

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Ren, T., Wang, Y., Wang, C. et al. Isolation and identification of human metabolites from a novel anti-tumor candidate drug 5-chlorogenic acid injection by HPLC-HRMS/MSn and HPLC-SPE-NMR. Anal Bioanal Chem 409, 7035–7048 (2017). https://doi.org/10.1007/s00216-017-0657-3

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  • DOI: https://doi.org/10.1007/s00216-017-0657-3

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