Compound B isolated from the crude extract of EL000327 exhibited cytotoxicity toward the human CRC cell line, Caco2. (a) Image of the endolichenic fungus, EL000327, belonging to Pseudoplectania sp. isolated from the lichen Graphis. (b) IC50 values of EL000327 in HT29, HCT116, Caco2, DLD1, CSC221, TMK1, RV1, A549, HaCaT, CT26, and MDCK cells. (c) Schematic representation of the process for the purification of compound B from the crude extract, EL000327. (d) Chemical structure of the novel compound, B. (e) IC50 values of human CRC cells Caco2, HCT116, DLD1, and HT29 after treatment with B for 48 h. (f) Comparison of IC50 values of the CRC cells Caco2 and non-cancer cell lines HaCaT and MDCK treated with single compound B, fraction 2, or crude extract, EL000327 for 48 h. Results are representative of three independent experiments. Data represent the mean ± S.D. * p < 0.05, ** p < 0.01, *** p < 0.001, NS: no significant difference (p > 0.05) compared with the Caco2 cells.