Demethoxycurcumin and rosmarinic acid as dual neuroprotective and anti-Epstein-Barr virus glycoprotein 350 agents

The Epstein-Barr virus (EBV) proteins EBNA1, LMP1, BZLF1, and gp350 have been consistently detected in the cerebrospinal fluid of patients with neurological disorders. Among these viral proteins, gp350 plays a critical role in determining viral tropism. In this study, phytocompounds were screened against the extra-virion domain of gp350. Based on their binding affinities, the top hits were subsequently subjected to 100-ns molecular dynamics simulations. Two phytocompounds, demethoxycurcumin (DMC) and rosmarinic acid (RA) were prioritized for subsequent in-vitro and in-vivo validation. Both compounds exhibited potent anti-gp350 activity in neuronal cells. Additionally, DMC- and RA-treated samples showed reduced levels of neuroinflammatory markers, including IL-6, TNF-α, NF-kB, and STAT3, compared with positive controls (acyclovir- or fingolimod-treated samples). Behavioral assays in mice revealed improved spatial memory in DMC- and RA-treated groups. Histological analyses demonstrated that EBV-infected mice displayed a disorganized hippocampal architecture relative to wild-type littermates, whereas hippocampal morphology was preserved in DMC- and RA-treated mice. Collectively, these findings indicate that DMC and RA exert significant anti-gp350 and neuroprotective effects and may hold potential as prophylactic and therapeutic candidates against EBV-associated neurodegeneration.