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Tricholoma anatolicum  is an edible mushroom from the matsutake group growing under Cedar trees. Bioactivity-guided fractionation of  Tricholoma anatolicum  afforded two new ( 1  and  2 ), three known ergosterols ( 3 – 6 ), and four known ( 6 – 9 ) compounds. Structures were identified as anatoluin A ( 1 ), anatoluin B ( 2 ), 5 α ,6 α -epoxy-ergosta-7,22-dien,3 β -ol ( 3 ), ergosterol-endoperoxide ( 4 ), ergosterol,3 β -ol ( 5 ), 3,5-dihydroxyfuran-2(5 H )-one ( 6 ), mannitol ( 7 ), turanose ( 8 ), fumaric acid ( 9 ) using spectroscopic techniques. The cytotoxic activity of extract and isolated compounds was performed using MTT assay against MCF7, HT29, H1299, and HeLa cancerous cell lines while toxicity against PDF and L929 fibroblast healthy cell lines. The lipid peroxidation inhibitory and ABTS •+  scavenging activities were used to determine antioxidant activity. The polar extracts exhibited significant cytotoxic activity. The more perfect is that the extracts and isolated compounds ( 1 – 5 ) were inactive against PDF and L929 healthy cell lines. Compounds  1 – 3  and  4  exhibited noticeable cytotoxic activity, while  1 – 5  moderately inhibited lipid peroxidation.

Anatoluin A and B isolated from medicinal Tricholoma anatolicum are new cytotoxic ergostanoids against the most common cancers