Synthesis and in vitro study of novel neuraminidase inhibitors against avian influenza virus.

Abstract

Evidences of oseltamivir resistant influenza patients raised the need of novel neuraminidase inhibitors. In this study, five oseltamivir analogs PMC-31-PMC-36, synthesised according to the outcomes of a rational design analysis aimed to investigate the effects of substitution at the 5-amino and 4-amido groups of oseltamivir on its antiviral activity, were screened for their inhibition against neuraminidase N1 and N3. The enzymes used as models were from the avian influenza A H7N1 and H7N3 viruses. The neuraminidase inhibition assay was carried out by using recombinant species obtained from a baculovirus expression system and the fluorogenic substrate MUNANA. The assay was validated by using oseltamivir carboxylate as a reference inhibitor. Among the tested compounds, PMC-36 showed the highest inhibition on N1 with an IC(50) of 14.6±3.0nM (oseltamivir 25±4nM), while PMC-35 showed a significant inhibitory effect on N3 with an IC(50) of 0.1±0.03nM (oseltamivir 0.2±0.02nM). The analysis of the inhibitory properties of this panel of compounds allowed a preliminary assessment of a structure-activity relationship for the modification of the 4-amido and 5-amino groups of oseltamivir carboxylate. The substitution of the acetamido group in the oseltamivir structure with a 2-butenylamido moiety reduced the observed activity, while the introduction of a propenylamido group was well tolerated. Substitution of the free 5-amino group of oseltamivir carboxylate with an azide, decreased the activity against both N1 and N3. When these structural changes were both introduced, a dramatic reduction of activity was observed for both N1 and N3. The alkylation of the free 5-amino group in oseltamivir carboxylate introducing an isopropyl group seemed to increase the inhibitory effect for both N1 and N3 neuraminidases, displaying a more pronounced effect against N1.

• Authors: Jarinrat Kongkamnerd, Luca Cappelletti, Adolfo Prandi, Pierfausto Seneci, Thanyada Rungrotmongkol, Nutthapon Jongaroonngamsang, Pornchai Rojsitthisak, Vladimir Frecer, Adelaide Milani, Giovanni Cattoli, Calogero Terregino, Ilaria Capua, Luca Beneduce, Andrea Gallotta, Paolo Pengo, Giorgio Fassina, Stanislav Miertus, Wanchai De-Eknamkul
• Journal: Bioorganic & medicinal chemistry (Bioorg Med Chem) Vol. 20 Issue 6 Pg. 2152-7 (Mar 15 2012) ISSN: 1464-3391 [Electronic] England
• PMID: 22342267 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012. Published by Elsevier Ltd.

Chemical References

• Antiviral Agents
• Oseltamivir
• Neuraminidase

Topics

• Animals
• Antiviral Agents (chemical synthesis, chemistry, pharmacology)
• Binding Sites
• Birds (virology)
• Influenza A Virus, H7N1 Subtype (chemistry, drug effects, enzymology)
• Influenza A Virus, H7N3 Subtype (chemistry, drug effects, enzymology)
• Influenza A virus (chemistry, drug effects, enzymology)
• Influenza in Birds (drug therapy, enzymology)
• Models, Molecular
• Neuraminidase (antagonists & inhibitors, chemistry, metabolism)
• Oseltamivir (analogs & derivatives, chemical synthesis, pharmacology)