Phosphorylated Neuropeptides Regulate Glioblastoma Proliferation and Invasion via Chiral Amino Acids

Neuropeptides are endogenous neuromodulators that have been shown to play pivotal roles in cancer initiation and progression. The phosphorylation/dephosphorylation processes are critically involved in regulating the bioactivity of peptides. This study designed chiral phosphorylated neuropeptides with distinct modification sites to examine the stereochemical regulatory mechanisms of alkaline phosphatase-mediated dephosphorylation and its biological effects on glioblastoma (GBM). The differential dephosphorylation catalyzed by alkaline phosphatase (ALP) demonstrates that both the spatial positions and quantity of modification sites significantly influence aggregation kinetics and intermolecular interactions. The dephosphorylated products derived from d-enantiomer probes inhibit GBM proliferation and invasion by disrupting cytoskeletal reorganization, while simultaneously enhancing tumor-targeted accumulation capacity and prolonging in vivo metabolic stability. These findings provide crucial insights into the stereochemical effects of chirality and terminal phosphorylation sites on peptide functionality, as well as contribute to the development for Y₁R-targeted therapies for GBM and other solid tumors.