Noregen(TM), a regenerative medicine ocular therapeutic

Project Summary Diabetic retinopathy and Retinal Vein OcclusionRVOaccount for most new cases of blindness in the United StatesThese diseases are marked by retinal swellingedemaand new leaky vessel formationneovascularizationtriggered by pathologically high concentrations of Vascular Endothelial Growth FactorVEGFThe primary treatment is VEGF blocking drugs to control the neovascularization and edemabut neuronal damage from oxygen starvation continues to progressSteroids treat edemaswellingbut do not address neovascularization and they have adverse side effectsSome VEGF is needed for neuronal and vascular endothelial cell healthso alternative treatments are needed to regenerate and repair the retinal vasculature without blocking normal VEGF signalingWe propose to use a recombinant protein mimetic of human Norrincalled Noregenas a safe regenerative therapy based on our patented technologyThere is an excellent scientific basis for this proposed regenerative therapyNorrin is a normal stimulator of retinal endothelial cells and is essential for making the retinal vasculatureNorrin also supports the repair and regrowth of blood vessels into retinal areas that have lost vessels and improves the organization and speed of vessel regrowth in a mouse model of oxygen induced retinopathyAdditionallyour current results demonstrate that Norrin can counter VEGF induced changes that increase the leakiness through endothelial cells caused by Plasmalemma Vesicle Associated ProteinPLVAPWhile elevated VEGF increases the expression of the PLVAP genewe have found that Norrin reduces PLVAP gene expressionIn Phase I we will develop a Noregen production process and conduct preclinical testing in animal models and with vascular endothelial cells from human donor retinasSuccessful completion of Phase I will provide the basic process for making the Noregen protein and the test methods needed to confirm that the protein is also effective for stimulating endothelial cell replacement and the formation of a vascular endothelial cell barrierIn Aimwe will establish bacterial production and purification of Noregen proteinwith Frizzledbinding activityRecombinant Noregen will be synthesized in bacteriarefolded into a biologically active state and purified using cation exchange chromatographyIn Aimwe will test Noregen protein s safety using a rat intraocular injection modelPurification steps will be added as required based on the results to reduce bacterial endotoxinIn Aimwe will confirm Noregen s efficacy to support regeneration of the retinal vasculature in a mouse model and maintenance of the endothelial barrier in cultured primary Human Retinal Endothelial CellsCompletion of Phase I will permit Retinal Solutions to submit a Phase II application where Noregen production will be transferred into a current Good Manufacturing ProcesscGMPfacilityand the therapeutic grade product used for additional pre clinical safety testing to meet the Food and Drug Administration requirements for intraocular drugs PROJECT NARRATIVE Diabetic retinopathy and Retinal Vein OcclusionRVOaccount for most new cases of blindness in the United StatesCurrent drug treatments slow vascular damage in the short term but eventually damage the neural retina long termIn this project we propose to use Noregena recombinant protein mimetic of the human growth factor Norrinto stimulate regeneration and repair of vascular damage as a safe and effective alternative therapy