Studies on the inhibition mechanism of the human coronavirus NL63 replication by polycations

Justyna Ciejka 1Aleksandra Milewska 2Kamil K. Kamiński 1Anna Karewicz 1Katarzyna Zasada 1Dorota Bielska 1Maria Nowakowska 1Jan Potempa 2Krzysztof Pyrć 2Krzysztof Szczubiałka 1

1. Jagiellonian University, Faculty of Chemistry, Ingardena 3, Kraków 30-060, Poland
2. Jagiellonian University, Faculty of Biochemistry, Biophysics and Biotechnology, Kraków 30-387, Poland


Human coronavirus NL63 (HCoV NL63) is generally classified as a common cold virus. It may cause severe upper and lower respiratory tract diseases. Thus, there is a need to develop effective methods of preventing and treating these infections.

The aim of this study was to synthesize and explore anticoronaviral activity of polymer-based compounds. Four polycations, i.e. N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC), hydrophobically-modified HTCC (HM-HTCC), O-(2-hydroxypropyl)-3-trimethylammonium poly(vinyl alcohol) chloride (HTPVA) and poly(allylamine hydrochloride) (PAH) were investigated.  In order to evaluate the inhibitory activity of the tested polymers in vitro study using LLC MK2 cells and ex vivo study using human airway epithelium (HAE) cultures were performed. The cytopathic effect (CPE) was correlated with a quantitative RT PCR based assay. The cytotoxicity was examined by an XTT assay and a Neutral Red assay.

The results show that cationically modified polymers (HTCC, HM-HTCC, PAH and HTPV) are effective inhibitors of human coronavirus NL63 replication. Evaluated polymers show relatively low cytotoxicity. What is more, their specificity is high. None of the compounds inhibited replication of other human viruses (influenza A, hMPV, adenoviruses, enteroviruses, and human herpes virus type 1).

Mechanism of antiviral activity is complicated. Size and charge of polymers suggest that they are not actively transported into the cell. Analysis of interaction between HTCC polymer and the recombinant ectodomain of the S protein showed binding, resulting in the formation of protein-polymer complexes.



Project operated within the Foundation for Polish Science Team Programme (PolyMed, TEAM/2008-2/6) and Ventures Programme (VENTURES/2013-11/1) co-financed by the EU European Regional Development Fund. Project partially financed by a grant from the Ministry of Scientific Research, Poland (0095/B/P01/2009/37 and N N204 151336), Iuventus Plus grant from the Ministry of Science and Higher Education, Poland (IP 2010 033870) (KP) and Foundation for Polish Science within the Homing Programme. 

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Presentation: Poster at Nano and Advanced Materials Workshop and Fair, by Justyna Ciejka
See On-line Journal of Nano and Advanced Materials Workshop and Fair

Submitted: 2013-06-29 15:20
Revised:   2013-08-13 22:28
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