Identification of novel small molecule inhibitors against NS2B/NS3 serine protease from Zika virus

• Published in: Antiviral research 2017-03, Vol.139 (C), p.49-58
• Main Authors: Lee, Hyun, Ren, Jinhong, Nocadello, Salvatore, Rice, Amy J., Ojeda, Isabel, Light, Samuel, Minasov, George, Vargas, Jason, Nagarathnam, Dhanapalan, Anderson, Wayne F., Johnson, Michael E.
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Antiviral Agents - chemistry; Antiviral Agents - pharmacology; apo ZIKV NS2B-NS3pro structure; BASIC BIOLOGICAL SCIENCES; Drug Discovery; Inhibitory Concentration 50; Kinetics; NS2B/NS3 serine protease; Protein Conformation; Serine Proteases - metabolism; Serine Proteinase Inhibitors - chemistry; Serine Proteinase Inhibitors - pharmacology; Small molecule inhibitor; Surface Plasmon Resonance; Viral Nonstructural Proteins - antagonists & inhibitors; Virus Replication - drug effects; Zika flavivirus; Zika Virus - drug effects; Zika Virus - enzymology
• ISSN: 0166-3542; 1872-9096
• DOI: 10.1016/j.antiviral.2016.12.016

Zika flavivirus infection during pregnancy appears to produce higher risk of microcephaly, and also causes multiple neurological problems such as Guillain–Barré syndrome. The Zika virus is now widespread in Central and South America, and is anticipated to become an increasing risk in the southern United States. With continuing global travel and the spread of the mosquito vector, the exposure is expected to accelerate, but there are no currently approved treatments against the Zika virus. The Zika NS2B/NS3 protease is an attractive drug target due to its essential role in viral replication. Our studies have identified several compounds with inhibitory activity (IC50) and binding affinity (KD) of ∼5–10 μM against the Zika NS2B-NS3 protease from testing 71 HCV NS3/NS4A inhibitors that were initially discovered by high-throughput screening of 40,967 compounds. Competition surface plasmon resonance studies and mechanism of inhibition analyses by enzyme kinetics subsequently determined the best compound to be a competitive inhibitor with a Ki value of 9.5 μM. We also determined the X-ray structure of the Zika NS2B-NS3 protease in a “pre-open conformation”, a conformation never observed before for any flavivirus proteases. This provides the foundation for new structure-based inhibitor design. [Display omitted] •We identified several small molecule inhibitors of the Zika NS2B-NS3 protease.•Two lead compounds were determined to be competitive inhibitors by both enzymatic and competition SPR binding analyses.•We determined the X-ray structure of the apo Zika NS2B-NS3 protease in a “pre-open conformation”.