Susceptibility of coxsackievirus B3 laboratory strains and clinical isolates to the capsid function inhibitor pleconaril: antiviral studies with virus chimeras demonstrate the crucial role of amino acid 1092 in treatment.
OBJECTIVE
At present, most promising compounds to treat enterovirus-induced diseases are broad-spectrum capsid function inhibitors which bind into a hydrophobic pocket in viral capsid protein 1 (VP1). Coxsackievirus B3 (CVB3) Nancy was the only prototypic enterovirus strain shown to be pleconaril-resistant. This study was designed to better understand the polymorphism of the hydrophobic pocket in CVB3 laboratory strains and clinical isolates and its implications for treatment with the capsid function inhibitor pleconaril.


METHODS
Pleconaril susceptibility was determined in cytopathic effect-inhibitory, plaque reduction or virus yield assays. Sequence analysis of the genome region coding for VP1 and/or subsequent alignment of amino acids lining the hydrophobic pocket of five CVB3 laboratory strains and 20 clinical isolates were carried out. Virus chimeras and computational analysis were used to prove the role of amino acid 1092.


CONCLUSIONS
Despite high conservation of pocket amino acids, polymorphism was detected at positions 1092, 1094 and 1180. Neither Pro-1094-->Thr nor Val-1180-->Ile altered efficacy of pleconaril treatment. But the amino acid at position 1092 was strongly associated with susceptibility of CVB3 to the capsid inhibitor. Whereas leucine was involved in resistance, isoleucine and valine were detected in pleconaril-susceptible CVB3. Results from antiviral assays with hybrid viruses demonstrate the crucial role of amino acid 1092 in pleconaril susceptibility. A resistant cDNA-generated CVB3 became pleconaril-susceptible after accepting parts from the genome region encoding Ile-1092 into its capsid. Computational analysis suggests that conformational changes in the hydrophobic pocket occur when leucine is substituted for isoleucine or valine and that this change leads to susceptibility to pleconaril.
Schmidtke
Michaela
M
Institute of Virology and Antiviral Therapy, Medical Centre of the Friedrich Schiller University Jena, Hans Knoell Str. 2, D-07740 Jena, Germany. michaela.schmidtke@med.uni-jena.de
Hammerschmidt
Elke
E
Schüler
Susanne
S
Zell
Roland
R
Birch-Hirschfeld
Eckhard
E
Makarov
Vadim A
VA
Riabova
Olga B
OB
Wutzler
Peter
P
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D016466:CHO Cells; D002213:Capsid / Q000187:drug effects* / Q000502:physiology; D036022:Capsid Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D024882:Drug Resistance, Viral; D029822:Enterovirus B, Human / Q000145:classification / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D004769:Enterovirus Infections / Q000821:virology; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D010069:Oxadiazoles / Q000494:pharmacology*; D010080:Oxazoles; D011110:Polymorphism, Genetic
D000998:Antiviral Agents; D036022:Capsid Proteins; D010069:Oxadiazoles; D010080:Oxazoles; C115201:pleconaril