title: Design of a Water Soluble Fluorescent 3-Hydroxy-4-Pyridinone Ligand Active at Physiological pH Values.
abstract: In the present work we report the structure and the spectroscopic characterization of a new fluorescent 3-hydroxy-4-pyridinone ligand D-3,4-HPO. The synthesis of the compound was performed in two steps, which involve the reaction of the commercially available fluorophore dansyl chloride with a 3-hydroxy-4-pyridinone chelating unit and further deprotection. The new fluorescent chelator was characterized in the solid state by single-crystal X-ray diffraction and in solution by NMR, MS, absorption and fluorescence spectroscopies. The analysis of the variation of the absorption spectrum with pH allowed the determination of four pK a values (pK a1  = 3.50, pK a2  = 4.50, pK a3  = 9.60, pK a4  = 10.20) and establishment of the corresponding distribution diagram. The study of the fluorescence properties of the ligand show that in the pH range between 4 and 9 the fluorescence intensity is constant and has its maximum value thus allowing its further use at physiological pH values. The interaction of the ligand with copper(II) was accessed by fluorescence spectroscopy in MOPS buffer and the results show that the presence of copper(II) quenches the fluorescence of the ligand in ca 94 % at a ligand: metal ratio of 2:1. The latter result is consistent with the formation of a copper(II) complex with the bidentate ligand, as confirmed by the EPR spectroscopy. Graphical Abstract New water soluble fluorescent ligand active at physiological pH values.
authors.affiliation: REQUIMTE-UCIBIO, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007, Porto, Portugal. acleite@fc.up.pt.
authors.forename: Andreia
authors.initials: A
authors.lastname: Leite
authors.affiliation: REQUIMTE-UCIBIO, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007, Porto, Portugal.
authors.forename: Ana M G
authors.initials: AM
authors.lastname: Silva
authors.affiliation: REQUIMTE-UCIBIO, Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, 4050-313, Porto, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Coutinho
authors.affiliation: REQUIMTE-LAQV, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007, Porto, Portugal.
authors.forename: Luís
authors.initials: L
authors.lastname: Cunha-Silva
authors.affiliation: REQUIMTE-LAQV, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007, Porto, Portugal.
authors.forename: Baltazar
authors.initials: B
authors.lastname: de Castro
authors.affiliation: REQUIMTE-UCIBIO, Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, 4050-313, Porto, Portugal.
authors.forename: Maria
authors.initials: M
authors.lastname: Rangel
keywords: 3-hydroxy-4-pyridinones; Copper II; Dansyl chloride; Fluorescence properties; Microwave-assisted synthesis