Efficient Discovery And Capturing Of New NNOS-PSD-95 Uncouplers From Herbal Medicines Using Magnetic Molecularly Imprinted Polymers As Artificial Antibodies

Stroke is the leading cause of adult disability,cognitive impairment and death.To disrupt the nNOS-PSD-95 interaction can be a better drug target for the prevention and treatment of neuronal damage resulting from ischemic insults.ZL006 is an efficient inhibitor of the nNOS-PSD-95 interaction with few or no adverse effects However,considering the strong hydrophilicity and unideal brain distribution of ZL006,more efficient nNOS-PSD-95 uncouplers are very imperative.To do so,highly selective magnetic molecularly imprinted polymers(MMIPs)with a core-shell structure were prepared as artificial antibodies.According to the results of computational simulations,we designed and synthesized MMIPs using ZL006 as template,AA as functional monomer and acetonitrile as solvent with the molar ratio of 1:5:25(ZL006:AA:EGDMA).Characterization and performance tests revealed that the most appropriate artificial antibodies showed uniform spherical morphologies,large adsorption capacities,fast binding kinetics,high selectivity and quick separation.These artificial antibodies were then used as sorbents for dispersive magnetic solid phase extraction(DSPE)coupled with HPLC and MS in order to capture and identify structural analogues to ZL006 from extracts of Scutellariae radix,Psoraleae fructus and Trifolium pratense.Furthermore,according to the neuroprotective effect and co-immunoprecipitation test,Baicalein,Neobavaisoflavone,Corylifol A and Biochanin A can be the potential uncouplers of nNOS-PSD-95.Therefore,this present study contributes valuable information for the discovery of nNOS-PSD-95 uncouplers from herbal medicines.