Potential multifunctional components explored in Chinese herbal medicine against hepatitis B combining multi-target affinity ultrafiltration HPLC/MS

Abstract

Background: Dysosma versipellis (D. versipellis) and Phyllanthus urinaria (P. urinaria) are two typical Chinese herbs with diverse chemical composition and affluent pharmacological activities. Yet, the specific bioactive components with anti-inflammatory and anti-viral effects and corresponding mechanisms remain unclear. Purpose: The aim of this study was to screen for the specific constituents with anti-inflammatory and anti-viral activities from the anti-HBV formulation. Study design: The anti-viral and anti-inflammatory activities of D. versipellis, P. urinaria and the anti-HBV formulation extracts were evaluated by in vitro ACE2 and COX-2 inhibitory assays. Affinity ultrafiltration combining liquid chromatography / mass spectrometry (UF-LC/MS) with COX-2, ACE2, and NTCP was conducted according to its empirical traditional use, and then molecular docking analysis was applied to validate the activity of the ligands. Methods: An efficient strategy was employed to screen bioactive compounds from the anti-HBV formulation. Corresponding activities of the extracts or phytochemicals were confirmed by inhibitor screening kits or docking simulation. Results: A total of 8, 3 and 8 phytochemicals exerting distinct affinities with COX-2, ACE2 or NTCP were rapidly screened out and characterized from D. versipellis, P. urinaria and the anti-HBV formulation, respectively. Thereafter, the interaction between the specific binding ligands, namely dehydropodophyllotoxin, β-apopicropodophyllin, quercetin, and COX-2, ACE2 or NTCP was further simulated by docking analysis, and the results were well correspondent with the UF results. Noticeably, the interaction network among multiple component-target-activity correlations was for the first time constructed in an effort to elucidate the mechanisms of action exerting its therapeutic effects for the anti-HBV formulation, which was also better in accordance with the traditional Chinese medicine (TCM) theory. Conclusion: The present study showcased an integrative strategy that could offer not only a puissant tool to screen for active constituents from this anti-HBV formulation, and further help to unveil its underlying mechanisms, but also provide an alternative platform for discovering new candidate drugs for anti-inflammation and anti-virus from other medicinal plants in the near future.

Keywords

• Dysosma versipellis
• Phyllanthus urinaria
• Anti-hepatitis B virus formulation
• Affinity ultrafiltration
• Multiple drug targets