One new anthraquinone derivative, named 8- O-methylnidurufin (1), together with five known analogues (2)(3)(4)(5)(6), have been isolated from a gorgonian-derived fungus, Aspergillus sp. The structures were elucidated by combined spectroscopic methods including 1D and 2D NMR spectral data. All isolated metabolites were evaluated for their cytotoxic and antibacterial activities in vitro. Compound 2 showed significant cytotoxic activity against K562 and HL-60 cell lines with IC 50 values of 0.87 and 1.46 PM, respectively.Marine-derived fungi of the genus Aspergillus are well known as a prolific source of structurally unique and biologically active metabolites. A growing number of novel bioactive secondary metabolites isolated from this genus have been reported recently [1, 2]. As part of our continuing efforts to discover bioactive metabolites from marine-derived fungi in the South China Sea, we isolated several anthraquinone dimers, bisabolane sesquiterpenoids, and benzylazaphilone derivatives with potent cytotoxic activity [3][4][5]. Recently, we have investigated the chemical compositions of the fermentation broth of an Asperigillus sp. strain isolated from a gorgonian Dichotella gemmacea. One new anthraquinone derivative, named 8-O-methylnidurufin (1), together with five known analogues, nidurufin (2) [6], averufin (3) [7], 8-O-methylaverufin (4) [8], averufanin (5) [9], and 8-O-methylaverufanin (6) [10, 11], have been obtained from the fungal culture. Herein we describe the isolation, structure elucidation, and biological activity of these compounds.Compound 1 was isolated as an orange powder. Its molecular formula of C 21 H 18 O 8 (13 degrees of unsaturation) was determined by HR-ESI-MS. Careful comparison of the 1 H and 13 C NMR data of 1 with those of nidurufin (2) [6] suggested that 1 was very similar to 2. The most obvious differences in their 1 H NMR spectra were the presence of a singlet signal at G 3.88 assignable to the methoxy group and a hydrogen-bonded hydroxy signal at G 13.94 in 1 instead of two hydrogen-bonded hydroxy signals at G 12.50 and 12.21 in 2. It can be deduced that 1 is the methylation derivative of 2. The methoxy group in 1 was also confirmed by the 13 C NMR spectrum with a carbon signal at G 56.2. In the HMBC spectrum, correlations from 8-OCH 3 to C-8 showed that the methoxy group is attached to C-8. Detailed assignments of protons and carbons for 1 were unambiguously accomplished by analysis of the COSY, HMQC, and HMBC data ( Table 1). The relative configuration of 1 was determined by combination of proton-proton coupling constant analysis and NOESY experiment.