title: Identification of novel small molecule inhibitors against NS2B/NS3 serine protease from Zika virus.
abstract: 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 Kivalue 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.
authors.affiliation: Novalex Therapeutics, Inc., 2242 W Harrison Suite 201, Chicago, IL 60612, USA.
authors.forename: Hyun
authors.initials: H
authors.lastname: Lee
authors.affiliation: Center for Biomolecular Science, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA.
authors.forename: Jinhong
authors.initials: J
authors.lastname: Ren
authors.affiliation: Center for Structural Genomics of Infectious Diseases (CSGID), Dept. of Biochemistry and Molecular Genetics, Northwestern University, Feinberg School of Medicine, 303 E. Chicago Ave., Chicago, IL 60611, USA.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Nocadello
authors.affiliation: Center for Biomolecular Science, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA; Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 833 S. Wood Street, IL 60612, USA.
authors.forename: Amy J
authors.initials: AJ
authors.lastname: Rice
authors.affiliation: Center for Biomolecular Science, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA; Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 833 S. Wood Street, IL 60612, USA.
authors.forename: Isabel
authors.initials: I
authors.lastname: Ojeda
authors.affiliation: Center for Structural Genomics of Infectious Diseases (CSGID), Dept. of Biochemistry and Molecular Genetics, Northwestern University, Feinberg School of Medicine, 303 E. Chicago Ave., Chicago, IL 60611, USA.
authors.forename: Samuel
authors.initials: S
authors.lastname: Light
authors.affiliation: Center for Structural Genomics of Infectious Diseases (CSGID), Dept. of Biochemistry and Molecular Genetics, Northwestern University, Feinberg School of Medicine, 303 E. Chicago Ave., Chicago, IL 60611, USA.
authors.forename: George
authors.initials: G
authors.lastname: Minasov
authors.affiliation: Center for Biomolecular Science, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA; Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 833 S. Wood Street, IL 60612, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Vargas
authors.affiliation: DNSK International, LLC, NEBA Building, 925 Sherman Avenue, Hamden, CT 06514, USA.
authors.forename: Dhanapalan
authors.initials: D
authors.lastname: Nagarathnam
authors.affiliation: Center for Structural Genomics of Infectious Diseases (CSGID), Dept. of Biochemistry and Molecular Genetics, Northwestern University, Feinberg School of Medicine, 303 E. Chicago Ave., Chicago, IL 60611, USA.
authors.forename: Wayne F
authors.initials: WF
authors.lastname: Anderson
authors.affiliation: Novalex Therapeutics, Inc., 2242 W Harrison Suite 201, Chicago, IL 60612, USA. Electronic address: mssjohnson@novalextherapeutics.com.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Johnson
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology*; D055808:Drug Discovery; D020128:Inhibitory Concentration 50; D007700:Kinetics; D011487:Protein Conformation; D057057:Serine Proteases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D020349:Surface Plasmon Resonance; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors*; D014779:Virus Replication / Q000187:drug effects; D000071244:Zika Virus / Q000187:drug effects* / Q000201:enzymology
chemical_list: D000998:Antiviral Agents; D015842:Serine Proteinase Inhibitors; D017361:Viral Nonstructural Proteins; D057057:Serine Proteases
keywords: NS2B/NS3 serine protease; Small molecule inhibitor; Zika flavivirus; apo ZIKV NS2B-NS3(pro) structure