Objective 5-Formyl-3-methylene-3,6-dihydropyridine (FMDP) is a novel compound derived from the reaction of amino acids with formaldehyde (FA) and acrolein (ACR), and its content in biscuits has been reported to be more than 40 times higher than that of free acrolein and formaldehyde. Given the structural similarity to FDP-lysine, an advanced glycation end product (AGE) derived from acrolein, this study aims to evaluate the potential safety risk of FMDPs by assessing their ability to stimulate the receptor for advanced glycation end products (RAGE).Methods Three FMDP compounds, each possessing a 5-formyl-3-methylene-3,6-dihydropyridine core structure, are synthesized by reacting formaldehyde and acrolein with alanine, asparagine, and threonine, respectively. Following purification, RAW264.7 macrophages are cultured with various concentrations of each FMDP compound. Cytotoxicity, RAGE expression, proteomic profiles, and the Keap1-Nrf2 signaling pathway are subsequently evaluated.Results All three FMDP compounds exhibit significantly lower cytotoxicity than acrolein and formaldehyde. However, FMDP treatment upregulates RAGE expression in RAW264.7 cells relative to the control group, with threonine-derived FMDP showing the strongest stimulatory effect, increasing RAGE expression by 60% at an exposure concentration of 300 μg/mL. Proteomic analysis identifies 69, 59, and 111 differentially expressed proteins in response to alanine-, asparagine-, and threonine-derived FMDP, respectively. Gene Ontology (GO) enrichment analysis indicates that FMDPs increase the expression of proteins associated with oxidative stress. Western blotting assays reveal that FMDPs induce oxidative stress through activation of the Keap1-Nrf2 signaling pathway, as evidenced by elevated Nrf2 levels, enhanced dissociation of Keap1 from Nrf2, and increased expression of oxidoreductases, including NQO1, HO-1, and GCLC.Conclusion The food-borne compounds FMDPs can stimulate RAGE and induce oxidative stress, highlighting the need for attention to their potential health risks.