Notoamides are a family of prenylated indole alkaloids with unusual ring systems and possessing a range of significant pharmaceutical activities. Based on LC-MS/MS and genome orientations, ten undescribed notoamide-type alkaloids namely sclerotiamides I-R were isolated from a marine gorgonian-derived fungus Aspergillus sclerotiorum LZDX-33-4. Their structures were determined by extensive spectroscopic data, in association with ECD data and single-crystal X-ray diffraction for configurational assignments. Bioassays resulted in sclerotiamide J along with five analogs possessing inhibitory effects against LDH and IL-1β expression in BV-2 cells. Further investigation revealed that sclerotiamide J significantly inhibited NLRP3 inflammasome activation and blocked NLRP3 inflammasome-induced pyroptosis via amelioration of mitochondria damage. In addition, sclerotiamide L exhibited potent inhibition against pathogenic Staphylococcus aureus ATCC 29213 with MIC value of 4.0 μM and the growth of MRSA T144 and Enterococcus faecalis ATCC 29212. This study extends the chemical diversity of notoamide-type alkaloids, and provides potential anti-inflammasome and antibacterial lead compounds for further structure optimization. © 2022