Molecular Targets of Corneal Antiangiogenesis

Dr. Kyung Bo Kim will prepare the following reagents: 1) affinity agent labeled withaferin A (biotin-withaferin A) and 2) Protac, withaferin A attached to an E3 ubiquitin ligase recognition motif. He will characterize intermediates and fmal chemical products by mass spectrometry and NMR. He will synthesize these two compounds and provide periodic updates by email and by phone conference regarding the progress. The established key milestones are: a) synthesis ofC-27withaferin A-glycine, b) purification of27-linker coupled withaferin A, and c) synthesis ofProtac moiety. Task 1: Dr. Kim will generate biotin-withaferin A to be completed by November, 2011. The compound will be sent to the PI's lab at UCHC upon its completion. Task 2: Dr. Kim will synthesize withaferin A-Protac which involves attachment ofHIF-la pentapeptide to C-27-derivatized withaferin A. The yields ofwithaferinA-Protac are anticipated to be in range of4-5 mg and this compound will be chemically characterized using analytical chemistry methods. Dr. Kim will complete synthesis by December, 2011 and is expected to provide the PI with the compound in January, 2012 pending no complications with the processes. Task 3: Dr. Cidambi Srinivasan will work with the PI to provide statistical assistance for data obtained from mouse angiogenesis and wound healing experiments. Dr. Srinivasan will also assist in data analysis and validation for the analysis.