title: Pre-Assembly of Near-Infrared Fluorescent Multivalent Molecular Probes for Biological Imaging.
abstract: A programmable pre-assembly method is described and shown to produce near-infrared fluorescent molecular probes with tunable multivalent binding properties. The modular assembly process threads one or two copies of a tetralactam macrocycle onto a fluorescent PEGylated squaraine scaffold containing a complementary number of docking stations. Appended to the macrocycle periphery are multiple copies of a ligand that is known to target a biomarker. The structure and high purity of each threaded complex was determined by independent spectrometric methods and also by gel electrophoresis. Especially helpful were diagnostic red-shift and energy transfer features in the absorption and fluorescence spectra. The threaded complexes were found to be effective multivalent molecular probes for fluorescence microscopy and in vivo fluorescence imaging of living subjects. Two multivalent probes were prepared and tested for targeting of bone in mice. A pre-assembled probe with 12 bone-targeting iminodiacetate ligands produced more bone accumulation than an analogous pre-assembled probe with six iminodiacetate ligands. Notably, there was no loss in probe fluorescence at the bone target site after 24 h in the living animal, indicating that the pre-assembled fluorescent probe maintained very high mechanical and chemical stability on the skeletal surface. The study shows how this versatile pre-assembly method can be used in a parallel combinatorial manner to produce libraries of near-infrared fluorescent multivalent molecular probes for different types of imaging and diagnostic applications, with incremental structural changes in the number of targeting groups, linker lengths, linker flexibility, and degree of PEGylation.
authors.affiliation: Department of Chemistry and Biochemistry, 236 Nieuwland Science Hall, University of Notre Dame , Notre Dame, Indiana 46556, United States.
authors.forename: Evan M
authors.initials: EM
authors.lastname: Peck
authors.affiliation: Department of Chemistry and Biochemistry, 236 Nieuwland Science Hall, University of Notre Dame , Notre Dame, Indiana 46556, United States.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Battles
authors.affiliation: Department of Chemistry and Biochemistry, 236 Nieuwland Science Hall, University of Notre Dame , Notre Dame, Indiana 46556, United States.
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Rice
authors.affiliation: Department of Chemistry and Biochemistry, 236 Nieuwland Science Hall, University of Notre Dame , Notre Dame, Indiana 46556, United States.
authors.forename: Felicia M
authors.initials: FM
authors.lastname: Roland
authors.affiliation: Department of Chemistry and Biochemistry, 236 Nieuwland Science Hall, University of Notre Dame , Notre Dame, Indiana 46556, United States.
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Norquest
authors.affiliation: Department of Chemistry and Biochemistry, 236 Nieuwland Science Hall, University of Notre Dame , Notre Dame, Indiana 46556, United States.
authors.forename: Bradley D
authors.initials: BD
authors.lastname: Smith
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000000981:diagnostic imaging; D016466:CHO Cells; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000494:pharmacology; D007259:Infrared Rays; D007700:Kinetics; D047028:Macrocyclic Compounds / Q000737:chemistry; D008422:Materials Testing; D051379:Mice; D008958:Models, Molecular; D008968:Molecular Conformation; D011092:Polyethylene Glycols / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D047028:Macrocyclic Compounds; D011092:Polyethylene Glycols