Nanofiltration (NF) membranes with negative surface charge are of critical significance for the efficient removal of anionic dyes from wastewater. In this work, a novel negatively charged NF membrane was prepared from a new sulfonamide monomer, 2,4,6-trimethyl-3,5-diaminobenzenesulfonamide (TMDBSA) and trimesoyl chloride (TMC) via interfacial polymerization. The unique diamino-sulfonamide structure of TMDBSA would be conducive to the formation of a poly (amide-sulfonamide) (PASA) layer with enhanced negative charge while maintaining the appropriate degree of crosslinking. The TMDBSA-TMC membrane prepared at optimized condition exhibits both high water permeability (19.6 L m−2h−1 bar−1) and superior rejections of anionic dyes with different molecular weights (e.g. 98.3% rejection for methyl blue), reveals that Donnan exclusion dominates the rejection mechanism. More importantly, benefiting from the strong negatively charged surface and PASA structure, the TMDBSA-TMC membrane shows separation stability for the removal of methyl blue in a broad pH range, even under a harsh acid environment. Meanwhile, the membrane has excellent antifouling performance and long-term stability during the filtration tests. The newly developed negatively charged NF membrane demonstrates great practical application potential for the removal of anionic dyes with small molecular weights.