Therapeutic Potential of WP9QY: Exploring Peptide Engineering to Target TNF-Alpha in Neuroblastoma and Mitigate Neuroinflammation

This study investigates the therapeutic potential of the peptide WP9QY and its modified analogues (S8) in mitigating neuroinflammation by targeting tumor necrosis factor-alpha (TNF-alpha), a pivotal mediator of inflammatory pathways. WP9QY was synthesized alongside structural variants incorporating acetylation and non-canonical amino acids to enhance stability and binding affinity. Fluorescence binding assays demonstrated that the S8 analogue displayed superior TNF-alpha binding affinity (Kd = 34.16) compared to WP9QY (Kd = 90.46). Stability assessments revealed that the acetylated peptides exhibited enhanced resistance to proteolytic degradation, particularly in biological environments. Biological evaluations in the SH-SY5Y cell line, a neural model, indicated that S8 had reduced cytotoxicity compared to WP9QY, with the acetylated modified variant showing the highest cell viability and lowest toxicity. MitoBrilliant staining and ROS analysis confirmed the peptides’ ability to preserve mitochondrial content and mitigate oxidative stress under inflammatory conditions. Furthermore, a notable reduction in PD-L1 expression was observed, particularly with the combination of WP9QY and S8, suggesting immunomodulatory effects. These findings establish that structural modifications to WP9QY improve its therapeutic profile by enhancing stability, reducing toxicity, and modulating inflammatory and oxidative pathways. This study contributes valuable insights into peptide-based therapies for neurodegenerative diseases and neuroinflammatory disorders, providing a foundation for future optimization of peptide therapeutics.