Pharmaceuticals and methods for treating hypoxia and screening methods therefor

Abstract

Light-generating fusion proteins having a ligand binding site and a light-generating polypeptide moiety and their use as diagnostics, in drug screening and discovery, and as therapeutics, are disclosed. The light-generating fusion protein has a feature where the bioluminescence of the polypeptide moiety changes upon binding of a ligand at the ligand binding site. The ligand may be, for example, an enzyme present in an environment only under certain conditions, e.g., ubiquitin ligase in a hypoxic state, such that the light-generating fusion protein is “turned on” only under such conditions.

1 - 14 . (canceled) 
     
     
         15 . A method of treating or preventing a hypoxic or ischemic related disorder in a subject, comprising administering to a subject in need thereof a compound which decreases prolyl hydroxylase expression or activity, such that said hypoxic or ischemic related disorder is treated, wherein the compound is identified by a method comprising the steps of:
 a) bringing into contact a prolyl hydroxylase, a HIF1α polypeptide moiety and a putative modulator compound under conditions where prolyl hydroxylase, in the absence of modulator, is capable of acting on said HIF1α polypeptide moiety; and   b) measuring the degree of prolyl hydroxylase inhibition caused by said modulator compound.   
     
     
         16 . The method of  claim 15 , wherein said compound is a prolyl hydroxylase antibody, or a nucleic acid that decreases the expression of a nucleic acid that encodes a prolyl hydroxylase polypeptide. 
     
     
         17 . (canceled) 
     
     
         18 . The method of  claim 15 , wherein the hypoxic or ischemic related disorder is an acute event selected from the consisting of myocardial infarction, stroke, cancer, and diabetes. 
     
     
         19 . (canceled) 
     
     
         20 . The method of  claim 15 , wherein the hypoxic or ischemic related disorder is a chronic event selected from the group consisting of deep vein thrombosis, pulmonary embolus, and renal failure. 
     
     
         21 . The method of  claim 15 , wherein the half life of HIF in said subject is increased compared to a subject not exposed to said compound. 
     
     
         22 - 44 . (canceled) 
     
     
         45 . A method of treating or preventing a hypoxic or ischemic related disorder in a subject, comprising administering to a subject in need thereof a compound which modulates prolyl hydroxylation of HIF, such that said hypoxic or ischemic related disorder is treated. 
     
     
         46 . The method of  claim 45 , wherein said compound decreases prolyl hydroxylation of HIF. 
     
     
         47 - 50 . (canceled) 
     
     
         51 . A method of treating or preventing a HIF-related disorder in a subject, comprising administering to a subject in need thereof a compound which modulates prolyl hydroxylation of HIF, such that said HIF-related disorder is prevented, reversed or stabilized. 
     
     
         52 . (canceled) 
     
     
         53 . The method of  claim 51 , wherein said compound decreases prolyl hydroxylation of HIF. 
     
     
         54 . The method of  claim 51 , wherein said HIF-related disorder is selected from the group consisting of myocardial infarction, stroke, cancer, and diabetes.