Streptomyces  sp. MTCC-25420 and Streptomyces sp. MTCC-25512 isolated from the Shivalik region of India, has been demonstrated for tetrahydroanthracene antibiotics including setomimycin production. Combination of specific physico-chemical conditions have led to the antibiotic complex formation (upto 5.0 g/L) in 2-10 days of fermentation with maximum production at 4-8 days in shake flask as well as 5L to 500L stirred tank bioreactors using modified production media in combination with additional carbon/nitrogen/minerals and elicitations with yields upto 800mg/L of are 9,9' bianthracene antibiotic setomimycin. Setomimycin significantly abrogated cancer cell proliferation and inhibited cancer cell migration and invasion in highly metastatic cancer cells (MiaPaca-2, MCF-7, HT-29 and HCT-116) of Pancreatic, Breast and Colorectal origin by down regulating ERK and MEK proteins which are the major regulators of cell proliferation, differentiation and apoptosis. Interestingly setomimycin upregulated pro-apoptotic protein Par-4 and downregulated anti-apoptotic protein BCL-2 in Colon as well as breast cancer cells HCT-116 and MCF-7 respectively. Strong affinity of setomimycin towards MEK protein has been confirmed by molecular docking studies and Western blot analysis.