A new cyclic tripeptide, novofumigatamide (1) has been isolated from Aspergillus novofumigatus CBS117520. The relative structure of 1 was established on the basis of spectroscopic and physico-chemical data and chemical investigations. The absolute structure of 1 was determined by using a Marfey's method. Compound 1 showed non-specific antifungal activities against some human pathogenic fungi and did not inhibit cell proliferation for any tumor cells.