Multi-Target Neuroprotection by dl-PHPB in APP/PS1 Mice: a proteomic analysis

Yingni Sun^1,2,3*Guoliang Bai⁴Kangmin Yang⁵Yong Feng⁶Hongmei Sun⁶Li Xian⁷Hongwei Gao¹

• ¹School of Life Sciences, Ludong University, Yantai, Shandong Province, China
• ²Department of Pharmacology, Chinese Academy of Medical Sciences, Beijing, China
• ³Beijing Handian Pharmaceutical Co. Ltd, Beijing, China
• ⁴National Center for Pediatric Cancer Surveillance, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, China
• ⁵Center for Molecular Cardiology, University of Zürich, Schliere, Switzerland
• ⁶Qingdao Huangdao District People 's Hospital, Qingdao, China
• ⁷Medicine and Pharmacy Research Center, Binzhou Medical University, Yantai, China

Dl-PHPB [potassium 2-(1-hydroxypentyl) benzoate] demonstrates robust neuroprotective effects in preclinical models of Alzheimer’s disease (AD), significantly ameliorating cognitive deficits and pathological hallmarks. However, the underlying mechanism remains largely unclear. The current study primarily focused on elucidating dl-PHPB’s neuroprotective mechanisms and identifying potential targets in preclinical AD models. Comparative proteomic analyses were performed on APP/PS1 mice orally administered either dl-PHPB (30 mg/kg) or vehicle daily for 3 months, alongside vehicle-treated wild-type (WT) non-transgenic littermates as controls. Total proteins were separated using two-dimensional difference gel electrophoresis, and differentially expressed protein spots were identified via LC‒MS/MS. Our results revealed 11 altered proteins in the cortex and 10 in the hippocampus between the WT and APP/PS1 groups treated with vehicle. Following dl-PHPB treatment, 12 differentially expressed proteins were identified in the cortex and 9 in the hippocampus of APP/PS1 mice. These proteins are primarily involved in energy metabolism, neuronal structure, protein trafficking, inflammatory and oxidative responses, and amyloid β (Aβ) and Tau processes, among which several proteins were validated as potential therapeutic targets. Notably, the expression levels of cofilin-2 and VDAC1 in APP/PS1 mice were restored to near-normal levels by the treatment with dl-PHPB, memantine, or donepezil, and further clinical validation is required to establish their utility as AD biomarkers for therapeutic efficacy.