Shallow and Deep-water <em>Ophiura</em> Species Produce a Panel of Chlorin Compounds with Potent Photodynamic Anticancer Activities

A Pacific brittle star Ophiura sarsii has previously been shown to produce a chlorin (3S,4S)-14-Ethyl-9-(hydroxymethyl)-4,8,13,18-tetramethyl-20-oxo-3-phorbinepropanoic acid (ETPA) (1) with potent phototoxic activities making it applicable to photodynamic therapy. Using extensive LC-MS metabolite profiling, molecular network analysis and targeted isolation with de novo NMR structure elucidation, we here identify five additional chlorin compounds from O. sarsii and its deep-sea relative O. ooplax: 10S-Hydroxypheophorbide a (2), Pheophorbide a (3), Pyropheophorbide a (4), (3S,4S,21R)-14-Ethyl-9-(hydroxymethyl)-21-(methoxycarbonyl)-4,8,13,18-tetramethyl-20-oxo-3-phorbinepropanoic acid (5), and (3S,4S,21R)-14-Ethyl-21-hydroxy-9-(hydroxymethyl)-4,8,13,18-tetramethyl-20-oxo-3-phorbinepropanoic acid (6). Chlorins 5 and 6 have not been previously reported in natural sources. Interestingly, low amounts of chlorins 1-4 and 6 could also be identified in a distant species, the basket star Gorgonocephalus cf. eucnemis, demonstrating that chlorins are produced by a wide spectrum of marine invertebrates of the class Ophiuroidea. Following purification of these major Ophiura chlorin metabolites, we discovered the high singlet oxygen quantum yield upon their photoinduction and their strong phototoxicity against a panel of cancer cell lines. These studies identify an arsenal of brittle star chlorins as powerful natural photosensitizers with potential photodynamic therapy applications.