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Proceedings of the 31 st European Peptide SymposiumMichal Lebl, Morten Meldal, Knud J. Jensen, Thomas Hoeg-Jensen (Editors)European Peptide Society, 2010Design, Synthesis and RNase Activity of Novel PeptidomimeticsAgainst Influenza VirusesL.S. Koroleva 1,2 , O.V. Morozova 1,3 , E.I. Isaeva 3 , L.M. Rustamova 4 ,V.M. Sabynin 4 , N.P. Schmeleva 4 , N.V. Gribkova 4 , and V.N. Silnikov 11 Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia;2 Novosibirsk State University, Novosibirsk, Russia; 3 Ivanovskiy Institute of Virology,Moscow, Russia; 4 Research Institute for Epidemiology and Microbiology,Minsk, Republic of BelarusIntroductionInfluenza virus remains one of the major causes of respiratory diseases. Vaccinationefficacy is limited because of high mutation rate of the influenza virus genome RNA.Therapy of influenza is currently based on blocking of the ion channels formed by M2 virusprotein (drugs of the adamantane groups: amantadine and rimantadine) and neuraminidaseinhibitors (zanamivir and ozeltamivir) and search of new antivirals is in progress. The aim:design and synthesis of novel peptidomimetic: Xaa-diamine-Xaa, where Xaa - Lys, Glu,Ser, His, Pro, Thr, Trp, Leu, as well as analysis of their cytotoxicity and inhibition ofinfluenza virus A and B.Results and DiscussionRecently a number of artificial peptide ribonucleases modeling known catalytic centers ofnatural RNases have been described [1-3]. RNA cleavage was shown to be more efficientin the presence of aliphatic hydrophobic fragment. However, a potential role of alkyl chainin the structure of the artificial RNases remains unclear. Interaction of hydrophobicresidues in peptides was suggested to result in structurization of RNase mimetics insolutions thus enhancing their ribonuclease activity. To prove this suggestion, symmetricaliphatic diamides containing natural amino acid residues have been synthesized. Thecompounds were shown to possess high ribonuclease activity with modeloligoribonucleotides and HIV-1 recombinant RNA fragment of 96 nucleotides long.Peptidomimetics were synthesized using method of activated esters and Boc-strategy. Thegeneral scheme is shown in Figure 1. The purity of the intermediates and the products ofthe reactions were monitored by TLC. The structures of the target compounds wereconfirmed by1 H NMR spectroscopy and MALDI-TOF mass-spectrometry. Thepeptidomimetics have high solubility in water and DMSO, all solutions are stable andretain their activity after storage at +4 o C for 12 months.Peptidomimetics were dissolved in water and added to culture medium of MDCK cellspermissive to influenza, adeno- and adeno-associated viruses without a serum at concentrations50, 25, 12.5, 6.25 and 3.125 µg/ml before or simultaneously with infection withinfluenza viruses A and B. Then the cellular monolayers were observed for 3 days.H 2 NiO R *O NHEDiPAOOO NOii CF 3COOH/CH 2Cl 2 or HCOOH for Boc-TrpiiiPd/C, [H]H 2 NRNH 2ONHiOHNO *RONHHNOfor Boc-Ser, Boc-Thr, Boc-Gln, Boc-Leu,Boc-Trp, di-Boc-Lys, di-Boc-HisRiiOH SerNNH HisRNH 2NH 2 LysOGluOHFig. 1. General scheme of synthesis of artificial ribonucleases.H 2 NH 2 N*ROONHNHHNOR*NHiiOOHNHNROfor Boc-Ser(Bzl), Boc-Glu(OBzl)OOH Thr GlnNH 2LeuiiiNHTrpOR*RNH 2NH 2336