Identification of novel glucocerebrosidase chaperone for potential treatment of Parkinson's disease: An approach using in silico virtual screening, molecular docking and molecular dynamics, and in vitro studies.

Glucocerebrosidase (GCase), a GBA1 gene-encoded lysosomal enzyme, is a risk factor for Parkinson's disease (PD). Chaperones that increase GCase activity can potentially be disease-modifying agents in PD. To date, none of the registered treatments has demonstrated disease-modifying effects. Thus, chaperones for GCase were identified using in-silico virtual screening, molecular property filtering, and molecular dynamics and validated by circular dichroism, FT-IR, and Raman spectroscopies. In-vitro enzyme kinetics, thermal denaturation assay (TDA), and cell-line model were used to test their potential for GCase In-silico investigation revealed four compounds as candidate chaperones with adequate brain penetrability and binding energy (BE). Of them, GC466 showed ideal chaperoning characteristics, including potent BE -8.92 ± 0.68 Kcal/mol and binding affinity (K i) 0.64 ± 0.12 μM against rGCase (Asp146, Phe265, and His329 residues) at pH 7.0 than at 4.5 (BE: −5.06 Kcal/mol, K i : not found). Spectroscopic results confirmed the stability of GCase by GC466. TDA determined its chaperoning behavior, signified by improved rGCase thermal stabilization with stabilization ratio of 10.20 at 10 μM. In addition, it demonstrated GCase restorative, neurorestorative, and ROS scavenging activity in 6-OHDA treated cell-line model. Therefore, the present study may offer a novel chaperone with the potential to be a disease-modifying agent for PD. • GC466 was identified as the chaperone, which binds rGCase with greater affinity at pH 7.0 than at pH 4.5. • TDA confirmed GC466's chaperoning action by stabilizing rGCase before denaturation. • Asp146, Phe265, His329, and Tyr331 residues play a vital role in the GC466-rGCase complex stabilization. • In-vitro pH-dependent CD, FT-IR, and Raman spectroscopies confirmed in-silico pH-dependent protein-ligand stabilization. • GC466 shows anti-PD activity in SHSY-5Y cell lines due to enhanced GCase and ROS scavenging activities. [ABSTRACT FROM AUTHOR]