Aim: To get enough quantity and quality of DL1106 and its analogue to evaluate their anti-tumor activity, the fermentation conditions of marine actinomycetes REL-3 were optimized.Methods: Estimating the effects of fermentation time and the pH value and salt concentration of fermentation broth on the yield, the fermentation conditions of REL-3 were optimized.After extracting the collection of the fermentation liquid with equal volume of acetic acid ethyl three times, the relative content of the DL1106 and its analogue in the concentrated extracts was detected by HPLC.