Hericium erinaceus is a species of mushroom with high nutritional value that is used mainly as food in tropical countries. Phytochemical study of H. erinaceus led to the isolation of an undescribed compound, named as hericium VN ( 1 ), together with nine known compounds, 1-(2-formyl-1-pyrrolyl)butanoic acid ( 2 ), herierin III ( 3 ), 5'-(methylthio)adenosine ( 4 ), adenosine ( 5 ), nicotinic acid ( 6 ), (22 E ,24 R )-5 α ,8 α -epidioxyergosta-6,9(11),22-trien-3 β -ol ( 7 ), 5 α ,8 α -peroxycerevisterol ( 8 ), (22 E ,24 R )-5 α ,8 α -epidioxy-egosta-6,22-diene 3- O - β -D-glucopyranoside ( 9 ), and cerevisterol ( 10 ) based on extensive analyses of HR-ESI-MS, 1D and 2D NMR spectra. The absolute configuration of compound 1 was determined by experimental combined with calculated electronic circular dichroism spectra. Compound 7 exhibited cytotoxic effects against brain tumor cell line CCF-STTG1 with the IC 50 value of 15.50 µM, compared to that of the positive control compound, doxorubicin, which showed IC 50 value of 15.84 µM.