A practical and rapid screening method for influenza virus neuraminidase inhibitors based on fluorescence detection

Abstract

A new analytical method for rapid screening of influenza virus neuraminidase inhibitors was established. The method is based on the principle that, given a certain amount of neuraminidase, the sample and the neuraminidase act in the microplate for a period of time, and the active neuraminidase that is not inhibited by the sample can generate a fluorescence value at a specific wavelength after binding to the substrate, and the rate of inhibition of neuraminidase by the sample can be calculated based on the actual detected fluorescence value. This newly developed method was used to screen and evaluate the in vitro anti-neuraminidase activity of 39 high-purity compounds contained in three traditional Chinese herbal medicines, and finally 25 compounds with strong activity were obtained. The newly established neuraminidase inhibitor analytical method has these advantages of practicality, rapidity, high sensitivity and low cost, and has a good value for promotion and application. This article newly establishes a rapid, sensitive, simple and practical screening method for influenza virus neuraminidase inhibitors, which is a great complement to the existing methods and has a good promotion and application value.

Keywords: Fluorescence detection; Influenza virus neuraminidase inhibitor; Molecular docking; Quality consistency; Traditional Chinese medicine.

Fig. 1

Schematic diagram of the anti-NA activity screening method

Fig. 2

Dose–effect curve formed by different test concentrations of peramivir and the corresponding inhibition rates against NA

Fig. 3

Dose–effect curves formed by different test concentrations of six representative compounds with their corresponding inhibition rates against NA

Fig. 4

Schematic diagram of the docking of amino acid residues of neuraminidase with some typical compounds. a peramivir; b kaempferide; c luteolin; d quercitrin