Thiacycloalkynes for Copper‐Free Click Chemistry

Almeida, Gabriela de; Sletten, Ellen M.; Nakamura, Hitomi; Palaniappan, Krishnan K.; Bertozzi, Carolyn R.

doi:10.1002/ange.201106325

Cited by 51 publications

(21 citation statements)

References 35 publications

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“…BARAC is in fact an interesting example of the fine balancing act between reactivity and stability that comes along with the development of cyclooctyne probes: BARAC displays a reaction rate constant of nearly 1 mol −1 s −1 (see Table 1 ), but unfortunately is inherently unstable and rapidly decomposes [ 28 ]. Two cyclooctyne probes difluorobenzocycloocytne (DIFBO [ 55 ]) and 3,3,6,6-tetramethylthiaheptyne (TMTH [ 5 ]) are yet more reactive than BARAC but cannot be isolated in pure form before rapid decomposition takes place (not depicted in Fig. 5 ).…”

Section: Speeding Up Spaacmentioning

confidence: 99%

Strain-Promoted 1,3-Dipolar Cycloaddition of Cycloalkynes and Organic Azides

2016

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A nearly forgotten reaction discovered more than 60 years ago—the cycloaddition of a cyclic alkyne and an organic azide, leading to an aromatic triazole—enjoys a remarkable popularity. Originally discovered out of pure chemical curiosity, and dusted off early this century as an efficient and clean bioconjugation tool, the usefulness of cyclooctyne–azide cycloaddition is now adopted in a wide range of fields of chemical science and beyond. Its ease of operation, broad solvent compatibility, 100 % atom efficiency, and the high stability of the resulting triazole product, just to name a few aspects, have catapulted this so-called strain-promoted azide–alkyne cycloaddition (SPAAC) right into the top-shelf of the toolbox of chemical biologists, material scientists, biotechnologists, medicinal chemists, and more. In this chapter, a brief historic overview of cycloalkynes is provided first, along with the main synthetic strategies to prepare cycloalkynes and their chemical reactivities. Core aspects of the strain-promoted reaction of cycloalkynes with azides are covered, as well as tools to achieve further reaction acceleration by means of modulation of cycloalkyne structure, nature of azide, and choice of solvent.

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Section: Speeding Up Spaacmentioning

confidence: 99%

Strain-Promoted 1,3-Dipolar Cycloaddition of Cycloalkynes and Organic Azides

2016

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“…Apart from the OCT skeleton, cyclononyne and cycloheptyne derivatives presented as new approaches. Thiacycloheptyne (TMTH) had the second-order rate constant k 2 of 4 M −1 ·s −1 , the fastest SPAAC so far reported [128,129]. However, TMTH couldn’t be equipped with a label, which hinders the further affinity purification or fluorescence detection after bioorthogonal reaction.…”

Section: Strain-promoted Azide-alkyne Cycloaddition (Spaac)mentioning

confidence: 99%

Development and Applications of the Copper-Catalyzed Azide-Alkyne Cycloaddition (CuAAC) as a Bioorthogonal Reaction

2016

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Abstract:The emergence of bioorthogonal reactions has greatly broadened the scope of biomolecule labeling and detecting. Of all the bioorthogonal reactions that have been developed, the copper-catalyzed azide-alkyne cycloaddition (CuAAC) is the most widely applied one, mainly because of its relatively fast kinetics and high efficiency. However, the introduction of copper species to in vivo systems raises the issue of potential toxicity. In order to reduce the copper-induced toxicity and further improve the reaction kinetics and efficiency, different strategies have been adopted, including the development of diverse copper chelating ligands to assist the catalytic cycle and the development of chelating azides as reagents. Up to now, the optimization of CuAAC has facilitated its applications in labeling and identifying either specific biomolecule species or on the omics level. Herein, we mainly discuss the efforts in the development of CuAAC to better fit the bioorthogonal reaction criteria and its bioorthogonal applications both in vivo and in vitro.

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“…Generation of hydroxyl radicals in proximity to DNA causes base damages and DNA strand breaks; this effect is proportional to the accessible surface areas of the hydrogen atoms of the DNA backbone (9). To note, alternative bioconjugation methods can be employed to avoid undesirable effects on DNA damage, such as difluorocyclooctyne coupling using strain-promoted alkyne-azide cycloaddition chemistry (37) or redox-protected reactions (38,39).…”

Section: Discussionmentioning

confidence: 87%

From “Click” to “Fenton” chemistry for 5‐bromo‐2′‐deoxyuridine determination

et al. 2013

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Ascorbic acid (AA) and copper have been increasingly employed in flow cytometry (FCM) and high content analysis (HCA) since the introduction of "click chemistry" as a non-destructive alternative to classical 5-bromo-2 0 -deoxyuridine (BrdU) immunodetection for DNA synthesis and proliferation assays. Mixtures of ascorbate and catalytic copper, under certain experimental conditions, act as oxidizing agent, catalyzing the formation of reactive hydroxyl radicals through hydrogen peroxides decomposition via Fenton reaction. We developed a procedure for BrdU incorporation detection based on the use of AA and cupric ions as DNA damaging agents. Optimal DNA damaging conditions were identified and found to provide results comparable with "click" 5-ethynyldeoxyuridine (EdU) cycloaddition approach and classical BrdU immunodetection. Scavenger agents were found to prevent hydroxyl-induced DNA damages, providing the proof-of-concept for the use of this procedure for DNA denaturation prior to BrdU detection. We demonstrated hydroxyl radicals' reaction to be readily applicable to HCA and FCM assays, for both classical BrdU immunostaining and EdU cycloaddition procedure. This technique was successfully employed for BrdU pulse-chase experiments and in multiparametric immunofluorescence assays for the simultaneous detection of labile phosphoproteins in intact cells. The use of AA/Cu prior to immunodetection for BrdU incorporation assays is a viable alternative to chemical/physical DNA denaturing agents (acids or heat), since it allows preservation of labile epitopes such as phosphoproteins, and over enzymatic agents (digestion with DNases) for its lower cost. V C 2013International Society for Advancement of Cytometry Key terms ascorbic acid; BrdU; click chemistry; Fenton chemistry; high content analysis L-Ascorbic acid (Vitamin C, AA) is a water-soluble antioxidant, which efficiently scavenges free radicals and other radical oxygen species (ROS) produced by cell metabolism. ROS generation is associated with several forms of tissue damage and plays a role in the development of a multitude of chronic diseases (1-3). AA acts as donor/acceptor in redox reactions, providing a protective activity against free radicals. A mechanism able to control ROS generated during aerobic metabolism is essential for cell viability and AA and glutathione work together to prevent accumulation of oxidative damages that can lead to cell death (4).Despite its role as radical scavenger, in the presence of free metal ions AA acts as a pro-oxidant and can damage biological molecules, such as nucleic acids. AA reduces cupric (Cu 11 ) to cuprous (Cu 1 ) ion, which is capable of catalyzing the formation of reactive hydroxyl radicals through hydrogen peroxides decomposition via the so-called Fenton's reaction (5-7). The hydroxyl radicals efficiently react with deoxyribose backbone of DNA (8): hydroxyl radicals abstract an hydrogen atom from deoxyribose, leading to the formation of a free radical intermediate, and eventually triggering extensive damages ...

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Thiacycloalkynes for Copper‐Free Click Chemistry

Cited by 51 publications

References 35 publications

Strain-Promoted 1,3-Dipolar Cycloaddition of Cycloalkynes and Organic Azides

Strain-Promoted 1,3-Dipolar Cycloaddition of Cycloalkynes and Organic Azides

Development and Applications of the Copper-Catalyzed Azide-Alkyne Cycloaddition (CuAAC) as a Bioorthogonal Reaction

From “Click” to “Fenton” chemistry for 5‐bromo‐2′‐deoxyuridine determination

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