title: Photocontrollable peroxynitrite generator based on N-methyl-N-nitrosoaminophenol for cellular application.
abstract: We designed and synthesized a photocontrollable peroxynitrite (ONOO(-)) generator, P-NAP, which has N-methyl-N-nitrosoaminophenol structure with four methyl groups introduced onto the benzene ring to block reaction of the photodecomposition product with ONOO(-) and to lower the semiquinoneimine's redox potential. The semiquinoneimine intermediate generated by photoinduced release of nitric oxide (NO) reduces dissolved molecular oxygen to generate superoxide radical anion (O(2)(•-)), which reacts with NO to afford ONOO(-) under diffusion control (k = 6.7 × 10(9) M(-1) s(-1)). NO release from P-NAP under UV-A (330-380 nm) irradiation was confirmed by ESR spin trapping. Tyrosine nitration, characteristic of ONOO(-), was demonstrated by HPLC analysis of a photoirradiated aqueous solution of P-NAP and N-acetyl-l-tyrosine ethyl ester. ONOO(-) formation was confirmed with a ONOO(-)-specific fluorogenic probe, HKGreen-3, and compared with that from 3-(4-morpholinyl)sydnonimine hydrochloride (SIN-1), which is the most widely used ONOO(-) generator at present. The photoreaction of P-NAP was influenced by superoxide dismutase, indicating that generation of O(2)(•-) occurs before ONOO(-) formation. The quantum yield for formation of duroquinone, the main P-NAP photodecomposition product, was measured as 0.86 ± 0.07 at 334 nm with a potassium ferrioxalate actinometer. Generation of ONOO(-) from P-NAP in HCT-116 cells upon photoirradiation was successfully imaged with HKGreen-3A. This is the first example of a photocontrollable ONOO(-) donor applicable to cultured cells.
authors.affiliation: Graduate School of Pharmaceutical Science, Nagoya City University, 3-1, Tanabe-dori, Mizuho-ku, Nagoya, Aichi 467-8603, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Ieda
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Nakagawa
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
authors.forename: Dan
authors.initials: D
authors.lastname: Yang
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Suzuki
authors.forename: Naoki
authors.initials: N
authors.lastname: Miyata
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry; D045325:HCT116 Cells; D006801:Humans; D015394:Molecular Structure; D009603:Nitroso Compounds / Q000737:chemistry*; D030421:Peroxynitrous Acid / Q000737:chemistry*; D010636:Phenols / Q000737:chemistry*; D055668:Photochemical Processes; D014407:Tumor Cells, Cultured
chemical_list: D005456:Fluorescent Dyes; C572029:N-methyl-N-nitrosoaminophenol; D009603:Nitroso Compounds; D010636:Phenols; D030421:Peroxynitrous Acid