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Graphical abstract Highlights • TTB is an effective anti-inflammatory agent to develop nutraceutical. • TTB suppresses NO production through inhibition of MAPKs/ NF-κB phosphorylation. • In zebrafish, TTB significantly reduced the production of NO and expression of iNOS. Abstract Brown algae are known to contain rich anti-inflammatory compounds. The objective of this study was to identify anti-inflammatory compound in the Sargassum macrocarpum and anti-inflammatory effect of tuberatolide B (TTB) was investigated. TTB significantly suppressed the production of NO and pro-inflammatory cytokines such as interleukin (IL)-6 and IL-1β. In addition, the mechanism involved in the inhibition of the inflammatory effect of LPS by TTB was investigated. The results showed that TTB aforementioned this effect through NF-κB and MAPKs phosphorylation pathways. We also assessed the anti-inflammatory activity of TTB using in vivo zebrafish model. In LPS-stimulated zebrafish, TTB enhanced survival and significantly inhibited the NO production and mRNA expression of inducible NO synthase. Therefore, anti-inflammatory activity of TTB against LPS in RAW264.7 cells and the zebrafish model was determined. These finding suggest that TTB may be used as functional anti-inflammatory foods and nutraceuticals. [ABSTRACT FROM AUTHOR]