Artemisia annua is the plant that produces
artemisinin,
an endoperoxide-containing sesquiterpenoid used for the treatment
of malaria. A. annua extracts, which contain other
bioactive compounds, have been used to treat other diseases, including
cancer and COVID-19, the disease caused by the virus SARS-CoV-2. In
this study, a methyl ester derivative of arteannuin B was isolated
when A. annua leaves were extracted with a 1:1 mixture
of methanol and dichloromethane. This methyl ester was thought to
be formed from the reaction between arteannuin B and the extracting
solvent, which was supported by the fact that arteannuin B underwent
1,2-addition when it was dissolved in deuteromethanol. In contrast,
in the presence of N-acetylcysteine methyl ester,
a 1,4-addition (thiol-Michael reaction) occurred. Arteannuin B hindered
the activity of the SARS CoV-2 main protease (nonstructural protein
5, NSP5), a cysteine protease, through time-dependent inhibition.
The active site cysteine residue of NSP5 (cysteine-145) formed a covalent
bond with arteannuin B as determined by mass spectrometry. In order
to determine whether cysteine adduction by arteannuin B can inhibit
the development of cancer cells, similar experiments were performed
with caspase-8, the cysteine protease enzyme overexpressed in glioblastoma.
Time-dependent inhibition and cysteine adduction assays suggested
arteannuin B inhibits caspase-8 and adducts to the active site cysteine
residue (cysteine-360), respectively. Overall, these results enhance
our understanding of how A. annua possesses antiviral
and cytotoxic activities.