The Beibu Gulf represents an underexplored reservoir of marine micro-/organisms and secondary metabolites. Three uncommon 4-hydroxy-2-pyridone alkaloids and one phenazine alkaloid were obtained from the Beibu Gulf coral-associated fungus Acremonium sclerotigenum GXIMD 02501 via OSMAC approach. They were identified as campyridones D (1) and A (2), ilicicolin H (3), and phenazine-1-carboxylic acid (4), respectively, by spectroscopic analysis and comparison with literature values. All of them were evaluated for cytotoxic, anti-Vibrio, and NF-kappa B luciferase inhibitory activities. Compounds 2 and 3 showed cytotoxicity against two prostate cancer cell lines, with IC50 values of 17.6 +/- 1.3 and 5.5 +/- 1.2 mu M for PC- 3, while 25.4 +/- 1.7 and 11.9 +/- 1.3 mu M for 22Rv1, respectively. Besides, compound 4 showed promising anti-Vibrio activity with MIC values of 0.047-0.067 mg/mL and also displayed inhibition of LPS-induced NF-.B activation at 10 mu M.