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Two new polyene polyketides, namely aurovertins T and U (1 and 2), were isolated from Calcarisporium arbuscula, together with aurovertins B (3), D and E (4 and 5), and M (6). The structures were elucidated by extensive spectroscopic methods (especially 2D‐NMR techniques). The cytotoxic activities of all isolates against human triple‐negative breast cancer cell line (MDA‐MB‐231) were evaluated. As a result, compounds 3, 4, and 6 exhibited more potent cytotoxic activities against MDA‐MB‐231 cell line than the positive control taxol. Also, discussion about the relationships between structure and activity of these aurovertins was presented.