Multivalent carbocyanine molecular probes: synthesis and applications

Bioconjug Chem. 2005 Jan-Feb;16(1):51-61. doi: 10.1021/bc049790i.

Abstract

Synergistic multivalent interactions can amplify desired chemical or biological molecular recognitions. We report a new class of multicarboxylate-containing carbocyanine dye constructs for use as optical scaffolds that not only serve as fluorescent antennas but also participate in structural assembly of the multivalent molecular construct. Three generations of carboxylate-terminating multivalent near-infrared carbocyanine probes from a dicarboxylic acid precursor dye (cypate) were prepared via its imino diacetic acid derivatives. Conjugation of the probes with D-(+)-glucosamine afforded dendritic arrays of the carbohydrates on an inner NIR chromophore core. All the multicarboxylate probes and their glucosamine conjugates have similar NIR spectral properties because conjugation occurred at distal positions to the inner chromophore core, thereby providing consistent and predictable spectral properties for their biological applications. Although light-induced photodamage equally affected the precursor dye, multicarboxylate probes, and their glucosamine derivatives, we observed that octacarboxylcypate (multivalent probe) was remarkably stable in different mediums at physiologically relevant temperatures relative to cypate, especially in basic mediums. Biodistribution studies in tumor-bearing nude mice show that all the glucosamine conjugates localized in the tumor but cypate was almost exclusively retained in the liver at 24 h postinjection. The tumor uptake does not correlate with the number of glucosamine tether on the multicarboxylate probe. Overall, the triglucosamine derivative appears to offer the best balance between high tumor uptake and low retention in nontarget tissues. These results suggest that multivalent molecular beacons are useful for assessing the beneficial effects of multivalency and for optimizing the biological and chemical properties of tissue-specific molecular probes.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Carbocyanines / chemical synthesis*
  • Chelating Agents / chemistry
  • Contrast Media / chemical synthesis*
  • Contrast Media / metabolism
  • Glucosamine / chemistry
  • Glycoconjugates / chemistry
  • Mice
  • Mice, Nude
  • Molecular Probes / chemical synthesis*
  • Neoplasm Transplantation
  • Neoplasms / diagnosis*
  • Spectrometry, Fluorescence
  • Spectroscopy, Near-Infrared
  • Tissue Distribution
  • Tumor Cells, Cultured

Substances

  • Carbocyanines
  • Chelating Agents
  • Contrast Media
  • Glycoconjugates
  • Molecular Probes
  • Glucosamine