Prodigiosin (1) is the parent member of a class of polypyrrole natural products that exhibit promising anticancer activities. They can facilitate copper-promoted oxidative DNA damage by binding to copper ions, and this activity is thought to represent their mechanism of cytotoxicity in the dark. They also possess photoinduced cytotoxicity, although 1 is too toxic in the dark to be used effectively for the treatment of cancer by photodynamic therapies. To circumvent dark toxicity by prodigiosins, the semi-synthetic analogue 2, in which the N-pyrrolic atoms of 1 are methylated to block copper coordination, and the synthetic phenyl analogues 3 and 4, which lack the copper-coordinating A-pyrrole ring of 1, were tested for their ability to inhibit colony formation of HL-60 cancer cells in the absence and presence of visible light (λ > 495 nm). Our results show that 2-4 lack cytotoxicity in the dark, but are able to inhibit colony formation of HL-60 cells following irradiation for 30 min. The synthetic derivative 4 exhibits photo-induced cytotoxicity similar to that of the natural product 1, demonstrating the potential use of prodigiosin-based compounds for treatment of cancers following irradiation with visible light.