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US 2012/0058502 A1Published application
Method for Identifying Protease Inhibitors
AssigneeVertex Pharmaceuticals Incorporated
InventorWilliam P. Taylor
Life of this patent
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Abstract
The invention describes a method for assaying HCV NS3 protease activity using an NS3•4A protease molecule. The invention also provides a method for screening and identifying modulators of NS3 protease.
Claims20
A method for measuring the activity of the Hepatitis C NS3 protease comprising:
- a) providing a sample comprising an isolated protein comprising SEQ ID NO:1;
- b) adding a peptide substrate based on the NS5A/NS5B cleavage site for Hepatitis C genotype 1a;
- c) separating the substrate and products on a reverse phase HPLC column;
- and
- d) quantifying the product peaks.
2 more elements
A method for identifying modulators of the Hepatitis C NS3 protease comprising
- a) providing a sample comprising a protein comprising SEQ ID NO:1;
- b) introducing a test compound to the sample;
- c) adding a peptide substrate based on the NS5A/NS5B cleavage site for Hepatitis C genotype 1a;
- d) separating the substrate and products on a reverse phase HPLC column;
- and
- e) quantifying the product peaks.
3 more elements
A method for determining whether a test compound modifies the activity of the NS3 protease or the activity of the NS4A cofactor comprising:
- a) providing a sample comprising a protein comprising SEQ ID NO:1;
- b) introducing a test compound to the sample;
- c) adding an NS4A cofactor peptide;
- d) adding a peptide substrate based on the NS5A/NS5B cleavage site for Hepatitis C genotype 1a;
- e) measuring first amounts of products by separating the substrate and products on a reverse phase HPLC column;
- and quantifying the product peaks;
- and
- f) comparing first amounts of products to second amounts of products, wherein the second amounts of products are measured in the absence of a NS4A cofactor peptide.
5 more elements
A kit comprising
- a) a sample comprising a protein containing SEQ ID NO:1 and
- b) a peptide substrate based on the NS5A/NS5B cleavage site for Hepatitis C genotype 1a.
Dependencies are inferred from the claim text.