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Two new prenylxanthone derivatives, asperidulins A (1) and B (2), along with a known emodin analogue (3) were isolated from an apple-derived fungus Aspergillus nidulans KIB-HACM-01. Their structures were elucidated by interpretation of HRMS, NMR, and comparisons of specific optical rotation. Asperidulin B (2) exhibited a moderate cytotoxicity against A549 and BEAS-2B with an IC50 values of 13.62 ± 0.41 and 11.27 ± 0.52 μM, and methyl-averantin (3) showed moderate cytotoxicities against all six tested cell lines (HL-60, A549, SMMC-7721, MDA-MB-231, SW480, BEAS-2B) with IC50 values ranging from 8.93 ± 0.56 to 35.27 ± 0.25 μM. [ABSTRACT FROM AUTHOR]