Abstract
ABSTRACTMembers of the Flaviviridae family constitute a severe risk to human health. Whilst effective drugs have been developed against the hepacivirus HCV, no antiviral therapy is currently available for any other viruses, including the flaviviruses dengue (DENV), West Nile and Zika viruses. The RNA-dependent RNA polymerase (RdRp) is responsible for viral replication and represents an excellent therapeutic target with no homologue found in mammals. The identification of compounds targeting the RdRp of other flaviviruses is an active area of research. One of the main factors hampering further developments in the field is the difficulty in obtaining high-quality crystal information that could aid a structure-based drug discovery approach. To address this, we have developed a convenient and economical 96-well screening platform. We validated the screen by successfully obtaining crystals of both native DENV serotype 2 and 3 RdRps under several conditions included in the screen. In addition, we have obtained crystal structures of RdRp3 in complex with a previously identified fragment using both soaking and co-crystallization techniques. This work will streamline and accelerate the generation of crystal structures of viral RdRps and provide the community with a valuable tool to aid the development of structure-based antiviral design.
Highlights
Flaviviridae are a family of enveloped, positive single stranded RNA viruses
No antivirals are currently available and vaccines are limited to yellow fever virus (YFV), Japanese encephalitis virus (JEV) and tick-borne encephalitis virus (TBEV)
The vaccine currently licensed for Dengue virus (DENV) (Dengvaxia, Senofi-Pasteur) only has limited efficacy against some DENV serotypes, and concerns have been raised over its administration to children and seronegative individuals (Aguiar et al, 2016)
Summary
The genus Flavivirus, of the Flaviviridae family, counts over 70 different viruses (Fields et al, 2007; Kuno et al, 1998), including Dengue virus (DENV), Japanese encephalitis virus (JEV), tick-borne encephalitis virus (TBEV), West Nile virus (WNV), yellow fever virus (YFV) and Zika virus (ZIKV). Infection with DENV, which is estimated to affect 390 million people annually (Bhatt et al, 2013), can lead to an ample range of clinical manifestations, from mild fever to fatal dengue shock syndrome (Rajapakse, 2011), while infection with ZIKV has recently been shown to be responsible for the sudden surge in the number of cases of microcephaly and neurological abnormalities in new-borns, and for several cases of Guillain-Barré syndrome (Dyer, 2015; Oliveira Melo et al, 2016). No antivirals are currently available and vaccines are limited to YFV, JEV and TBEV. In the absence of safe and effective vaccines, and given the risk of emergence of new flaviviruses, as demonstrated by the recent re-emergence of ZIKV, the development of antivirals against this group of viruses becomes ever more important
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
