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Updated: Oct 18, 2025

A Fluorogenic Peptide Cleavage Assay to Screen for Proteolytic Activity: Applications for coronavirus spike protein activation

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Published on: January 9, 2019

Hypericin Inhibit Alpha-Coronavirus Replication by Targeting 3CL Protease.

Yue Zhang¹, Huijie Chen^1,2, Mengmeng Zou¹

¹Veterinary Pathology Laboratory, College of Veterinary Medicine, Heilongjiang Key Laboratory for Animal and Comparative Medicine, Northeast Agricultural University, Harbin 150030, China.

|September 28, 2021

Summary

Hypericin effectively inhibits porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) by targeting the 3CLpro enzyme. This compound shows promise as a broad-spectrum antiviral for alphacoronaviruses (α-CoVs).

Keywords:

3CL protease PEDV TGEV hypericin α-CoV

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Area of Science:

Virology
Biochemistry
Drug Discovery

Background:

Porcine epidemic diarrhea virus (PEDV), an Alphacoronavirus (α-CoV), causes significant economic losses in swine farming due to high piglet mortality.
Hypericin, a natural dianthrone compound, exhibits known antiviral properties against various viruses.
The 3CLpro enzyme is a crucial target for antiviral therapies against coronaviruses.

Purpose of the Study:

To investigate the antiviral efficacy of hypericin against PEDV.
To elucidate the mechanism of hypericin's action against PEDV, specifically its interaction with the viral 3CLpro enzyme.
To explore the potential of hypericin as a pan-anti-α-CoV therapeutic agent.

Main Methods:

In vitro antiviral assays to assess hypericin's effect on PEDV replication and egress.
Molecular docking simulations to predict hypericin's binding to PEDV 3CLpro.
Fluorescence resonance energy transfer (FRET) assays to confirm hypericin's inhibition of 3CLpro activity.
Sequence and structural alignments of α-CoV 3CLpro enzymes.
Validation of hypericin's anti-transmissible gastroenteritis virus (TGEV) activity.

Main Results:

Hypericin significantly reduced PEDV replication and egress.
Molecular docking revealed hypericin binds to two specific pockets in PEDV 3CLpro.
FRET assays confirmed hypericin's inhibition of PEDV 3CLpro activity.
Conserved binding pockets were identified in other α-CoVs, including TGEV.
Hypericin demonstrated antiviral effects against TGEV.

Conclusions:

Hypericin is a potent inhibitor of PEDV and TGEV, acting by targeting the 3CLpro enzyme.
The conserved binding sites suggest hypericin's potential as a broad-spectrum antiviral against α-CoVs.
Hypericin warrants further investigation as a therapeutic candidate for PEDV, TGEV, and potentially other coronavirus infections.