Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

Ya-Hsin Hsu; Yu-Ling Hsu; Sheng-Hung Liu; Hsin-Chia Liao; Po-Xuan Lee; Chao-Hsiung Lin; Lee-Chiang Lo; Shu-Ling Fu

doi:10.1371/journal.pone.0152770

Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

PLoS One. 2016 Apr 1;11(4):e0152770. doi: 10.1371/journal.pone.0152770. eCollection 2016.

Authors

Ya-Hsin Hsu¹, Yu-Ling Hsu², Sheng-Hung Liu^{3

4}, Hsin-Chia Liao¹, Po-Xuan Lee¹, Chao-Hsiung Lin⁵, Lee-Chiang Lo², Shu-Ling Fu^{1

4}

Affiliations

¹ Institute of Traditional Medicine, National Yang-Ming University, Taipei, Taiwan.
² Department of Chemistry, National Taiwan University, Taipei, Taiwan.
³ Department and Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan.
⁴ Program in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei, Taiwan.
⁵ Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Taipei, Taiwan.

Abstract

Andrographolide (ANDRO) is a lactone diterpenoid compound present in the medicinal plant Andrographis paniculata which is clinically applied for multiple human diseases in Asia and Europe. The pharmacological activities of andrographolide have been widely demonstrated, including anti-inflammation, anti-cancer and hepatoprotection. However, the pharmacological mechanism of andrographolide remains unclear. Therefore, further characterization on the kinetics and molecular targets of andrographolide is essential. In this study, we described the synthesis and characterization of a novel fluorescent andrographolide derivative (ANDRO-NBD). ANDRO-NBD exhibited a comparable anti-cancer spectrum to andrographolide: ANDRO-NBD was cytotoxic to various types of cancer cells and suppressed the migration activity of melanoma cells; ANDRO-NBD treatment induced the cleavage of heat shock protein 90 (Hsp90) and the downregulation of its client oncoproteins, v-Src and Bcr-abl. Notably, ANDRO-NBD showed superior inhibitory effects to andrographolide in all anticancer assays we have performed. In addition, ANDRO-NBD was further used as a fluorescent probe to investigate the uptake kinetics, cellular distribution and molecular targets of andrographolide. Our data revealed that ANDRO-NBD entered cells rapidly and its fluorescent signal could be detected in nucleus, cytoplasm, mitochondria, and lysosome. Moreover, we demonstrated that ANDRO-NBD was covalently bound to several putative target proteins of andrographolide, including NF-κB and hnRNPK. In summary, we developed a fluorescent andrographolide probe with comparable bioactivity to andrographolide, which serves as a powerful tool to explore the pharmacological mechanism of andrographolide.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult
Diterpenes / chemistry*
Female
Humans
Magnetic Resonance Imaging
Male
Middle Aged
Molecular Probes / chemistry*

Substances

Diterpenes
Molecular Probes
andrographolide

Grants and funding

Funded by Ministry of Science and Technology in Taiwan (https://www.most.gov.tw/en/public); Grant number: MOST-102-2113-M-002-004-MY3, MOST 103-2320-B-010-007-MY3, MOST 104-3011-B-010-001 and MOST-104-2311-B-010-005. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.