<jats:p> One new cytochalasin daldinin (1), 2 known cytochalasins 2 and 3, along with 2 steroids 4 and 5, were characterized from the methanol extracts of the fruiting bodies of Daldinia concentrica. The structure of new compound 1 was elucidated using a combination of 1- and 2-dimensional nuclear magnetic resonance spectroscopic and high-resolution electrospray ionization mass spectrometric analyses. In addition, the isolated compounds were examined for their cytotoxicity against several tumor cell lines and the tested compounds demonstrated moderate-to-weak cytotoxicity.