The identification and characterization of small molecules for the inhibition of NGF and proNGF8p75NTR interactions using surface plasmon resonance spectroscopy

Abstract

! The!survival!of!neurons!in!both!the!central!and!peripheral!nervous!systems! depend!on!a!balance!between!pro8survival!and!pro8death!signaling!pathways.! Neurotrophins!and!their!precursors,!proneurotrophins,!are!a!unique!family!of! soluble!signaling!proteins,!which!act!to!preserve!this!balance.!Dysregulation!of!these! proteins,!however,!has!been!implicated!in!several!pathologies.!Specifically,!both! nerve!growth!factor!(NGF)!and!pro8nerve!growth!factor!(proNGF)!have!been! implicated!in!the!development!and!progression!of!neurodegenerative!diseases,!such! as!Parkinson’s!and!Alzheimer’s!diseases,!through!p75NTR8mediated!apoptotic! signaling.!!Therefore,!the!identification!of!small!molecule!inhibitors!capable!of! binding!to!and!modulating!NGF!and!proNGF!may!be!of!therapeutic!interest.!This! thesis!aims!to!characterize!the!inhibitory!action!of!previously!reported!small! molecule8based!NGF!inhibitors,!ALE80540,!PD90780,!Ro!0882750,!and!PQC!083,!as! well!as!novel!derivatives!of!these!compounds,!using!surface!plasmon!resonance! (SPR)!spectroscopy.!Of!the!established!inhibitors,!PD90780!offered!the!most! effective!inhibitory!action!for!both!NGF!and!proNGF!binding!to!p75NTR,!suggesting! that!this!compound!may!offer!multipotent!inhibition.!Further,!several!novel!ligand! specific!molecules!were!identified,!which!selectively!inhibited!either!NGF!or! proNGF8p75NTR!interactions.!Specifically,!BVNP!1,!BVNP!6,!and!BVNP!7!were!shown! to!be!effective!inhibitors!of!NGF8p75NTR!interactions,!while!BVNP!3!and!BVNP!4! were!found!to!be!selective!for!the!inhibition!of!proNGF!binding!to!p75NTR.!The! findings!of!this!thesis!offer!new!insight!into!the!use!of!small!molecule!inhibitors!to! ! iv! block!both!neurotrophin!and!proneurotrophin8mediated!signaling.!In!addition,!the! compounds!described!herein!may!be!used!for!future!lead!development.!