This is a collaborative proposal to develop inhibitors with reduced susceptibility to resistance and improved genotype spectrum of activity against the NS3/4A protease of the Hepatitis C virus (HCV). Over three percent of the world's population is infected with the hepatitis C virus (HCV). Unfortunately, the current best treatment is still quite challenging against the most prevalent genotype 1. The HCV NS3/4A protease is an attractive drug target due to its essential role in viral replication. In prior work, we discovered two novel scaffolds that show inhibitory activity against four of the most prevalent genotypes, one of which shows an IC50 value of ~2 �M against wild type NS3 genotype 1b, and also maintained its potency within a 10-fold range against five drug-resistant mutants. Another, with an IC50 of ~2-20 �M against the more common genotypes will provide a backup hit. We propose to develop these scaffolds into a low molecular weight NS3/4A inhibitor with broader spectrum activity and fewer side effects than current therapeutics. We will pursue this goal through three targeted aims to (1) utilize scaffold expansion of current hits to develop more extensive Structure Activity Relationships (SARs) to increase potency by at least an order of magnitude; (2) use structure-based design and synthesis to further improve inhibitors; and (3) utilize metabolic stability and related pharmacokinetic parameters, along with HCV antiviral efficacy to iteratively improve inhibitor design therapeutic characteristics. With these Aims, we expect to attain Milestone criteria for success that include: reducing the inhibitor enzymatic IC50 to � 10 nM; obtaining antiviral EC50 � 100 nM (replicon assay); retaining good enzymatic selectivity for NS3/4A vs. other off-target enzymes; mouse microsomal stability, t1/2 > 30 min; potential for good oral bioavailability; and minimal cytotoxicity, with a selectivity index, SI � 100.
Public Health Relevance Statement: Public Health Relevance: Hepatitis C virus (HCV) is a major cause of chronic liver diseases and hepatocellular carcinoma, affecting more than 180 million people (~3% of world population). There are 3-4 million new infections each year, with more than 350,000 annual deaths from HCV related liver diseases. Our research will utilize a combination of computational chemistry, enzymology, X-ray crystallography, synthetic medicinal chemistry, mutational resistance analysis and preclinical biological studies to develop inhibitors of an enzyme that is essential for Hepatitis C infection and replication. We are developing these as a new potential antiviral therapeutic for the treatment of Hepatitis C.
Project Terms: Active Sites; Address; Adverse effects; Affect; analog; Antiviral Agents; base; Binding (Molecular Function); Biological; Biological Assay; Biological Availability; Cells; Cessation of life; Characteristics; chronic liver disease; Clinical Trials; computational chemistry; Coupled; Crystallization; cytotoxicity; design; Development; Docking; Drug Kinetics; Drug resistance; Drug Targeting; Enzymatic Biochemistry; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; experience; flexibility; Genotype; Goals; Hepatitis C; Hepatitis C virus; Hot Spot; improved; indexing; Infection; inhibitor/antagonist; Inhibitory Concentration 50; Interferons; Lead; Liver diseases; Metabolic; Methods; Modification; Molecular Structure; Molecular Weight; Mus; mutant; novel; Oral; Penetration; Peptide Hydrolases; Pharmaceutical Chemistry; Population; Positioning Attribute; pre-clinical; Predisposition; Primary carcinoma of the liver cells; Process; Protease Inhibitor; public health relevance; Quantitative Structure-Activity Relationship; Replicon; Research; Resistance; Ribavirin; Role; Route; scaffold; small molecule libraries; Solvents; Structure; Structure-Activity Relationship; success; System; Therapeutic; Vendor; Vertebral column; Viral; Work; X-Ray Crysta
