Strain Promoted Click Chemistry of 2- or 8‑Azidopurine and 5‑Azidopyrimidine Nucleosides and 8‑Azidoadenosine Triphosphate with Cyclooctynes. Application to Living Cell Fluorescent Imaging

• Published in: Bioconjugate chemistry 2015-08, Vol.26 (8), p.1519-1532
• Main Authors: Zayas, Jessica, Annoual, Marie, Das, Jayanta Kumar, Felty, Quentin, Gonzalez, Walter G, Miksovska, Jaroslava, Sharifai, Nima, Chiba, Akira, Wnuk, Stanislaw F
• Format: Article
• Language: English
• Subjects: Adenosine triphosphatase; Adenosine Triphosphate - analogs & derivatives; Adenosine Triphosphate - chemistry; Alkynes - chemistry; Aqueous solutions; Azides - chemistry; Biochemistry; Cell Membrane Permeability; Cell Proliferation; Cellular biology; Click Chemistry; Cyclooctanes - chemistry; Fluorescence; Fluorescent Dyes - chemistry; Humans; MCF-7 Cells; Microscopy, Fluorescence; Nucleosides - chemistry; Pyrimidines - chemistry; Signal transduction; Temperature effects; Triazoles - chemistry
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/acs.bioconjchem.5b00300

Strain-promoted click chemistry of nucleosides and nucleotides with an azido group directly attached to the purine and pyrimidine rings with various cyclooctynes in aqueous solution at ambient temperature resulted in efficient formation (3 min to 3 h) of fluorescent, light-up, triazole products. The 2- and 8-azidoadenine nucleosides reacted with fused cyclopropyl cyclooctyne, dibenzylcyclooctyne, or monofluorocyclooctyne to produce click products functionalized with hydroxyl, amino, N-hydroxysuccinimide, or biotin moieties. The 5-azidouridine and 5-azido-2′-deoxyuridine were similarly converted to the analogous triazole products in quantitative yields in less than 5 min. The 8-azido-ATP quantitatively afforded the triazole product with fused cyclopropyl cyclooctyne in aqueous acetonitrile (3 h). The novel triazole adducts at the 2- or 8-position of adenine or 5-position of uracil rings induce fluorescence properties which were used for direct imaging in MCF-7 cancer cells without the need for traditional fluorogenic reporters. FLIM of the triazole click adducts demonstrated their potential utility for dynamic measuring and tracking of signaling events inside single living cancer cells.