Secondary Metabolites with Antithrombotic and Antioxidant Activities Derived from Cordyceps cicadae

<i>Cordyceps cicadae</i>, a medicinal and edible entomopathogenic fungus, has been widely used in traditional Chinese medicine for treating various ailments. This study aimed to validate its ethnopharmacological uses by investigating bioactive constituents and their antithrombotic and antioxidant activities. Through various chromatographic separations, one unreported flavonoid; quercetin-3-<i>O</i>-<i>β</i>-D-methylglucopyranoside (<b>1</b>); three known flavonoids (<b>2</b>-<b>4</b>); and one new dicarboxylic acid derivative, cicadae acid (<b>5</b>), were isolated from <i>C</i>. <i>cicadae</i>. Their chemical structures were elucidated by a comprehensive spectroscopic analysis (1D/2D NMR and HRESIMS), electronic circular dichroism (ECD) calculations, a DP4+ probability analysis, and the modified Mosher method. All compounds exhibited significant antithrombotic effects at a concentration of 20 μM in a zebrafish model. Compounds <b>1</b>-<b>4</b> exhibited potent antioxidant activity in the DPPH radical scavenging assay, with IC₅₀ values ranging from 12.81 ± 3.42 to 20.16 ± 2.64 μM. These findings provide scientific evidence supporting the traditional application of <i>C. cicadae</i>, identifying specific flavonoids and dicarboxylic acids as potential therapeutic agents for thrombosis and oxidative stress-related disorders.