Memantine and amantadine KLVFF peptide conjugates: Synthesis, structure determination, amyloid-β interaction and effects on recognition memory in mice

Abstract

Background

Adamantane derivatives, such as memantine (Mem) and amantadine (Ada), have distinct mechanisms and therapeutic applications. Ada is primarily utilized as an antiviral and anti-Parkinson drug without significant pro-cognitive effects, Mem is effective in various clinical conditions characterized by cognitive deficits, including Alzheimer's disease. Recent evidence highlights a neuroprotective role for Aβ monomers, suggesting that preventing their aggregation into toxic oligomers could be a promising therapeutic strategy. Based on the observation that the Lys-Leu-Val-Phe-Phe (KLVFF) peptide, can block the transition of randomly coiled Aβ monomers into toxic β-sheet aggregates, two KLVFF conjugates, the Mem-Succ-KLVFF and Ada-Succ-KLVFF were investigated.

Methods

Peptides were synthesized by Microwave-Assisted Solid Phase Peptide Synthesis (MW-SPPS). Circular Dichroism (CD), Th-T fluorescence and Gel-Electrophoresis techniques were used to assess the inhibitory effect on Aβ₄₂ fibrillogenesis. The formation of inclusion complexes with β-Cyclodextrin (β-CyD) was demonstrated by NMR Spectroscopy. The Novel Object Recognition (NOR) test, followed by double-blind analysis, was applied for in vivo response to compounds administration. In vitro effects on neurons were studied by MTT assay and WB analysis, whereas HR ESI-MS allowed the molecular detection on brain homogenates.

Results

These compounds differently affect Aβ₄₂ aggregation. Mem-Succ-KLVFF, and Succ-KLVFF affect pCREB levels in differentiated SH-SY5Y, a signaling pathway involved in memory processes. In the NOR test, both Mem and KLVFF exhibited pro-cognitive effects individually and synergistically when co-administered.

Conclusion

Structure-activity relationships are discussed, integrating in vivo results, memory-related cellular pathways, and HR-ESI-MS analyses. These findings support the therapeutic potential of these compounds in preserving cognitive function.