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. 2008 Jul 17;10(14):3097-9.
doi: 10.1021/ol801141k. Epub 2008 Jun 13.

A hydrophilic azacyclooctyne for Cu-free click chemistry

Ellen M Sletten  1 , Carolyn R Bertozzi
Affiliations

A hydrophilic azacyclooctyne for Cu-free click chemistry

Ellen M Sletten et al. Org Lett. .

Abstract

Biomolecules labeled with azides can be detected through Cu-free click chemistry with cyclooctyne probes, but their intrinsic hydrophobicity can compromise bioavailability. Here, we report the synthesis and evaluation of a novel azacyclooctyne, 6,7-dimethoxyazacyclooct-4-yne (DIMAC). Generated in nine steps from a glucose analogue, DIMAC reacted with azide-labeled proteins and cells similarly to cyclooctynes. However, its superior polarity and water solubility reduced nonspecific binding, thereby improving the sensitivity of azide detection.

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Figures

Figure 1
Figure 1
Cu-free click chemistry with (A) cyclooctyne reagents or (B) 6,7-dimethoxyazacyclooct-4-yne (DIMAC) reagents.
Scheme 1
Scheme 1. Synthesis of DIMAC
Figure 2
Figure 2
(A) Biotin conjugates of DIMAC (7) and a parent cyclooctyne (8).(5a) (B) Western blot of Jurkat cell lysates. Cells were treated with (+Az) or without (−Az) 25 μM Ac4ManNAz for 3 d, and lysates (35 μg total protein) were reacted with 250 μM 7 or 8 overnight at 25 °C. Equal protein loading was confirmed by Ponceau S staining (Figure S2, Supporting Information). The blot was probed with HRP-conjugated α-biotin. (C) Flow cytometry analysis of Jurkat cells. Cells metabolically labeled as in (B) were reacted with 250 μM 7 or 8 for 1 h at 25 °C and then stained with FITC−avidin. Similar data were obtained in three replicate experiments.
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