Pharmaceutical agents for treating HCV infections

A method of screening for a pharmaceutical agent for treating a HCV infection, wherein the method comprises:

• (a) contacting a cell with a test compound;
• (b) measuring the amount of 3-ketodihydrosphingosine, dihydrosphingosine, dihydroceramide, or ceramide synthesized in the cell in the presence of the test compound;
• (c) selecting a test compound that reduces the amount of 3-ketodihydrosphingosine, dihydrosphingosine, dihydroceramide, or ceramide synthesized in the presence of the test compound in (b), as compared to a measured amount of 3-ketodihydrosphingosine, dihydrosphingosine, dihydroceramide, or ceramide, respectively, synthesized when the test compound is absent;
• and
• (d) assaying whether the test compound selected in (c) has inhibitory activity against HCV replication.

A method of screening for a pharmaceutical agent for treating a HCV infection, wherein the method comprises:

• (a) providing an enzyme in the palmitoyl CoA to sphingomyelin biosynthetic pathway;
• (b) contacting the enzyme with a test compound and a substrate of the enzyme;
• (c) measuring the amount of a product synthesized by the enzyme over time in the presence of the test compound;
• (d) determining that the test compound reduces the amount of the product synthesized by the enzyme over time in the presence of the test compound in (c), as compared to a measured amount of the product synthesized by the enzyme when the test compound is absent;
• and
• (e) assaying whether the test compound has inhibitory activity against HCV replication.

A method of screening for a pharmaceutical agent for treating a HCV infection, wherein the method comprises:

• (a) contacting an enzyme in the palmitoyl CoA to sphingomyelin biosynthetic pathway with a test compound and a substrate of the enzyme;
• (b) measuring the activity of the enzyme;
• (c) selecting a test compound that reduces the activity of the enzyme in the presence of the test compound in (b), as compared to a measured activity of the enzyme when the test compound is absent;
• and
• (d) assaying whether the test compound selected in (c) has inhibitory activity against HCV replication.

A method of screening for a pharmaceutical agent for treating a HCV infection, wherein the method comprises:

• (a) contacting an enzyme in the palmitoyl CoA to sphingomyelin biosynthetic pathway with a test compound and a substrate of the enzyme;
• (b) measuring the activity of the enzyme;
• (c) selecting a test compound that reduces the activity of the enzyme in the presence of the test compound in (b), as compared to a measured activity of the enzyme when the test compound is absent;
• and
• (d) providing a pharmaceutical agent for treating a HCV infection, the pharmaceutical agent comprising the selected test compound.

A method of screening for a pharmaceutical agent for treating a HCV infection, wherein the method comprises:

• (a) contacting a cell with a test compound;
• (b) measuring the amount of a product synthesized in the process of sphingomyelin biosynthesis from palmitoyl CoA in the cell in the presence of the test compound;
• (c) selecting a test compound that reduces the amount of the product synthesized in the presence of the test compound in (b), as compared to a measured amount of the product synthesized when the test compound is absent;
• and
• (d) providing a pharmaceutical agent for treating a HCV infection, the pharmaceutical agent comprising the selected test compound.

A method of screening for a pharmaceutical agent for treating a HCV infection, wherein the method comprises:

• (a) contacting a cell with a test compound;
• (b) measuring the amount of 3-ketodihydrosphingosine, dihydrosphingosine, dihydroceramide, or ceramide, respectively, synthesized in the cell in the presence of the test compound;
• (c) selecting a test compound that reduces the amount of 3-ketodihydrosphingosine, dihydrosphingosine, dihydroceramide, or ceramide synthesized in the presence of the test compound in (b), as compared to a measured amount of 3-ketodihydrosphingosine, dihydrosphingosine, dihydroceramide, or ceramide, respectively, synthesized when the test compound is absent;
• and
• (d) providing a pharmaceutical agent for treating a HCV infection, the pharmaceutical agent comprising the selected test compound.

A method of screening for a pharmaceutical agent for treating a HCV infection, wherein the method comprises:

• (a) providing an enzyme in the palmitoyl CoA to sphingomyelin biosynthetic pathway;
• (b) contacting the enzyme with a test compound and a substrate of the enzyme;
• (c) measuring the amount of a product synthesized by the enzyme over time in the presence of the test compound;
• (d) selecting a test compound that reduces the amount of the product synthesized by the enzyme over time in the presence of the test compound in (c), as compared to a measured amount of the product synthesized by the enzyme when the test compound is absent;
• and
• (e) providing a pharmaceutical agent for treating a HCV infection, the pharmaceutical agent comprising the selected test compound.