Novel Water-Soluble Near-Infrared Cyanine Dyes:  Synthesis, Spectral Properties, and Use in the Preparation of Internally Quenched Fluorescent Probes

• Published in: Bioconjugate chemistry Vol. 18; no. 4; pp. 1303 - 1317
• Main Authors: Bouteiller, Cédric, Clavé, Guillaume, Bernardin, Aude, Chipon, Bertrand, Massonneau, Marc, Renard, Pierre-Yves, Romieu, Anthony
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.07.2007
• Subjects: Amino acids; Biochemistry; Carbocyanines - chemical synthesis; Carbocyanines - chemistry; Carbocyanines - metabolism; Caspase 3 - metabolism; Chemical reactions; Chromatography, High Pressure Liquid; Fluorescence; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - chemistry; Fluorescent Dyes - metabolism; Infrared radiation; Peptides; Quenching; Solubility; Spectrum Analysis; Water - chemistry
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/bc0700281

In this paper, we describe the synthesis and the photophysical properties of two novel near-infrared (NIR) cyanine dyes (NIR5.5-2 and NIR7.0-2) which are water soluble potential substitutes of the commercially available Cy 5.5 and Cy 7.0 fluorescent labels respectively. For each one of these cyanine dyes, the synthetic strategy relies on the postsynthetic derivatization of a cyanine precursor in order to introduce the key functionalities required for bioconjugation of these NIR fluorophores. For NIR5.5-2, a reactive amino group was acylated with an original trisulfonated linker for water solubility. For NIR7.0-2, a vinylic chlorine atom was derivatized through a SRN1 reaction for the introduction of a monoreactive carboxyl group for labeling purposes. Unexpectedly, when these two fluorophores were closely associated within a peptidic architecture, mutual fluorescence quenching between NIR5.5-2 and NIR7.0-2 was observed both at 705 (NIR5.5-2) and 798 nm (NIR7.0-2). On the basis of this property, a novel internally quenched caspase-3-sensitive NIR fluorescent probe was prepared.