Cytotoxic biotransformed products from andrographolide by Rhizopus stolonifer ATCC 12939

Biotransformation of andrographolide (1) by Rhizopus stolonifer ATCC 12939 was investigated. Ten bioconversion products were isolated and identified. Their structures were elucidated by high resolution mass spectroscopy (HR-MS), extensive NMR techniques, including 1H NMR, 13C NMR, DEPT, 1H–1H correlation spectroscopy (COSY), two dimensional nuclear Overhauser effect correlation spectroscopy (NOESY), heteronuclear multiple quantum coherence (HMQC) and heteronuclear multiple bond coherence (HMBC). Their structures were identified to be 12(R),13(R)-12-hydroxyandrographolide (2), 12(S),13(S)-12-hydroxyandrographolide (3), isoandrographolide (4), 3-dehydro-isoandrographolide (5), 14-deoxy-11,12-didehydroandrographolide (6), 3-oxo-14-deoxy-11,12-didehydroandrographolide (7), 3-dehydroandrographolide (8), 14-deoxyandrographolide (9), 3-dehydro-14-deoxyandrographolide (10) and 3-dehydro-14-deoxyandrographolide-19-oic acid (11). Among them, compounds 5 and 11 are novel compounds. The biosynthetic pathways of andrographolide by R. stolonifer were proposed. Most of the products showed potential antiproliferative activities against human breast cancer (MCF-7), human colon cancer (HCT-116) and human leukemia (HL-60) cell lines, and their structure–activity relationships (SAR) were discussed in detail.