Assays, methods and means
AssigneeIsis Innovation Limited
InventorsPatrick Henry Maxwell, Christopher William Pugh, Peter John Ratcliffe, Christopher Joseph Schofield
Life of this patent
Estimating the term…
Abstract
A novel class of hydroxylases is described having the amino acid sequence of SEQ ID NO: 2, 4, 6 and 8, and variants and fragments thereof having HIF hydroxylation activity. The polypeptides of the invention have in particular prolyl hydroxylase activity. An assay method monitors the interaction of the HIF hydroxylase with a substrate. Modulators of HIF hydroxylase are provided for use in the treatment of a condition associated with increased or decreased HIF levels or activity or for the treatment of a condition where it is desirable to modulate HIF levels or activity.
Claims32
1-55. (canceled)
A method of inhibiting the activity of a human HIF hydroxylase in mediating the hydroxylation of one or more proline residues of a human HIF-α protein, comprising
- providing a substance identified as inhibiting such activity by assays comprising:
- contacting a HIF prolyl hydroxylase and a substrate of that hydroxylase under conditions in which the hydroxylase interacts with the substrate, in the presence or absence of a test substance;
- and
- determining the interaction, or lack of interaction of, the hydroxylase and the substrate, wherein the interaction of the hydroxylase with the substrate in the presence or absence of the test substance is determined by measuring the hydroxylase activity of the hydroxylase;
- wherein the HIF hydroxylase is chosen from (a) the amino acid sequence of SEQ ID NO:
- 2, 4, 6 or 8, FLJ21620 (BAB15101) or Clorfl2 (NP071334);
- (b) a variant thereof having at least 60% identity to the amino acid sequence of SEQ ID NO:
- 2, 4, 6 or 8 and having HIF hydroxylase activity;
- or (c) a fragment of either (a) or (b) having HIF hydroxylase activity.
Dependencies are inferred from the claim text.