Screening novel anti-inflammatory peptides inhibiting the NLRP3 inflammasome from UHPLC-MS/MS-characterized peptide profiles of cocoa tea protein hydrolysates

Cocoa tea (Camellia ptilophylla) is a popular beverage enjoyed worldwide, yet its protein-rich residues are often underutilized during processing. In this study, proteins from cocoa tea were extracted and hydrolyzed with alkaline protease to produce cocoa tea protein hydrolysates (CTPH). The results showed that CTPH exhibited promising anti-inflammatory activity. To uncover bioactive peptides, the hydrolysates were analyzed using UHPLC-MS/MS, and potential peptides were screened through molecular docking targeting the NLRP3 inflammasome. In an in vitro model of NLRP3 inflammasome activation, two active peptides, LLR and LIGF, were found to significantly reduce IL-1β production in PMA-differentiated THP-1 macrophages following treatment with nigericin or ATP. These peptides interact with the NACHT domain of NLRP3 via hydrogen bonding and hydrophobic interactions. Their binding to NLRP3 was further confirmed by CETSA assay. These findings suggest that cocoa tea-derived peptides could offer therapeutic potential for managing inflammation.