During search for novel bioactive materials from natural resources with the potential as health food and alternative medicine, the culture broth of Cordyceps longissima (CL) J106, J144 and C. scarabaeicola (CS) J94, J123 were prepared, and their effect on cytotoxicity and nitric oxide (NO) production in RAW 264.7 cells were investigated. Whereas the culture broth of CL J144 and CS J123 had cytotoxicity in RAW 264.7 cells, that of CL J106 and CS J94 did not. The culture broth of CL J106 and CS J94 suppressed NO production in RAW 264.7 cells activated with lipopolysaccharide (LPS) at a dose-dependent manner. These results suggest that culture broth, a by-product of Cordyceps, may have active compounds with anti-inflammatory effect. In addition, it appears that their biological activity is dependent on the strains in spite of the same species.