论文部分内容阅读
Aim It has been widely accepted that autophagy plays a key role in some human diseases such as Parkinson s disease (PD).UNC5 llike kinasesl (ULK1) has been widely reported to initiate autophagy via its complex ULK1mAtg13FIP200 at the first stage; however, targeting ULK1 as a therapeutic strategy in PD still remains in its infancy.This study aimed at developing a novel ULK1 activator as candidate drugs for PD therapy and validating the possible mechanism and efficacy in vitro and in vivo.Methods Sequence alignment, phylogenetic analysis, homology modeling, molecular dockingand structure modificationwere applied forscreening of candidate compounds.Surface plasmon resonance (SPR) analysis and molecular dynamics (MD) simulations were carried outto prove the binding betweenULK1 and BLUA07.Observations of cell morphology were executed through several methods including MDC staining and GFPLC3 transfection.Flow cytometric analysis of MDC was used for quantification of autophagy ratio.Western blot and RNAi transfection were used to explore the detailed mechanisms of BLUA07induced autophagy.Furthermore, an in vivo PD mouse model was established for validating the PD treatment efficacy of BLUA07.Results After a series of screening and structure modification, a novel compound BLUA07targeting ULK1 was obtained, which couldeffectivelybind with its target.Then, our results showed that BLUA07could induce autophagy via ULK1 complex and decrease damage induced by MPP + in SHSY5Y cells.In addition,in vivomouse model was established to evaluate the protective effect of BLUA07.The results demonstrated that BLUA07 has a therapeutic effect on the in vivomouse model without apparent toxicity, which is dependent on the cytoprotective autophagy mediated by ULK1.Conclusion In this study, a novel specific ULK1 activator (BLUA07)was computationally designed, chemically synthesized and biologically validatedthat could induce cytoprotective autophagy in neuroblastoma SHSY5Y cells and in vivo mouse models.Together, these results may uncover this smallmolecule compound BLUA07 as a novel ULK1 activator in autophagy and thus would provide a new clue for exploring more candidate drug targeting ULK1 for future PD therapy.