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Cytochalasans have continuously aroused considerable attention among the chemistry and pharmacology communities due to their structural complexities and pharmacological significances. Sixteen structurally diverse chaetoglobosins, 10-(indol-3-yl)-[13]cytochalasans, including a new one, 6-<i>O</i>-methyl-chaetoglobosin Q (<b>1</b>), were isolated from the coral-associated fungus <i>Chaetomium globosum</i> C2F17. Their structures were accomplished by extensive spectroscopic analysis combined with single-crystal X-ray crystallography and ECD calculations. Meanwhile, the structures and absolute configurations of the previously reported compounds <b>6</b>, <b>12</b>, and <b>13</b> were confirmed by single-crystal X-ray analysis for the first time. Chaetoglobosins E (<b>6</b>) and Fex (<b>11</b>) showed significant cytotoxicity against a panel of cancer cell lines, K562, A549, Huh7, H1975, MCF-7, U937, BGC823, HL60, Hela, and MOLT-4, with the IC₅₀ values ranging from 1.4 &#956;M to 9.2 &#956;M.