Screening Mitochondria-targeting Compounds from Traditional Chinese MedicinesUsing a Mitochondria-based Centrifugal Ultrafiltration/Liquid Chromatography/Mass Spectrometry Method

• 1、Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing 100191
• 2、 StateKey Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191
• 3、 College of Pharmaceutical Science, Yunnan University of Traditional Chinese Medicine, Kunming 650500
• 4、StateKey Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191
• 5、College of Pharmaceutical Science, Yunnan University of Traditional Chinese Medicine, Kunming 650500

Abstract： Mitochondria are becoming of increasing interest as a highly significant target for various disease remedy over the years, because mitochondria play a central role in energy production, apoptotic cell death, oxidative stress, calcium homeostasis and lipidic metabolism. We thus applied an efficient mitochondria-based centrifugal ultrafiltration/liquid chromatography/mass spectrometry method, which is called screening method for mitochondria-targeted bioactive constituents (SM-MBC) and is agree with the search of mitochondria-targeting compounds from complex matrix, to hunt for natural mitochondria-targeting compounds from traditional Chinese medicines (TCMs) including Puerariae Radix (PR) and Chuanxiong Radix (CR). Totally 23 active compouds were successfully discovered from the two extracts of TCMs. Among the 23 hit compounds, 17 were identified by LC/MS, of which 12 were new mitochondria-targeting compounds. The abilities of 7 of the 17 identified active compounds to remedy mitochondrial functions were exposed by in vitro pharmacological tests. Furthermore, the inhibition capabilities of the hit compounds on HepG2 cell proliferation and on loss of cardiomyocyte viability induced by hypoxia/reoxygenation injury were first exposed by cell tests. The results obtained are benefit for deep-going elucidation of the therapeutic principles of TCMs used to remedy mitochondria-involved disorders and drug development using TCMs as lead compounds, and SM-MBC was further showed to be an efficient protocol for the rapid screening of mitochondria-targeting constituents from complex preparation such as PR and CR extracts.

Keywords： Mitochondria Bioactive constituents Traditional Chinese medicines Centrifugal ultrafiltration Liquid chromatography/mass spectrometry