Tetrazine-trans-cyclooctene ligation for the rapid construction of integrin αvβ3 targeted PET tracer based on a cyclic RGD peptide

Selvaraj, Ramajeyam; Liu, Shuanglong; Hassink, Matthew; Huang, Chiun Wei; Yap, Li Peng; Park, Ryan; Fox, Joseph M.; Li, Zibo; Conti, Peter S.

doi:10.1016/j.bmcl.2011.04.116

Cited by 92 publications

(65 citation statements)

References 29 publications

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“…Although initial work focused on in vitro labeling there has been a surge of recent work applying this reaction for various in vivo applications (15)(16)(17). Despite the above progress, our results with initial TCO/Tz reactions in live animals were disappointing unless we used excessive amounts of reactants.…”

mentioning

confidence: 87%

Reactive polymer enables efficient in vivo bioorthogonal chemistry

Devaraj

Thurber

Keliher

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

179

187

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There has been intense interest in the development of selective bioorthogonal reactions or "click" chemistry that can proceed in live animals. Until now however, most reactions still require vast surpluses of reactants because of steep temporal and spatial concentration gradients. Using computational modeling and design of pharmacokinetically optimized reactants, we have developed a predictable method for efficient in vivo click reactions. Specifically, we show that polymer modified tetrazines (PMT) are a key enabler for in vivo bioorthogonal chemistry based on the very fast and catalyst-free [4 þ 2] tetrazine/trans-cyclooctene cycloaddition. Using fluorescent PMT for cellular resolution and 18 F labeled PMT for whole animal imaging, we show that cancer cell epitopes can be easily reacted in vivo. This generic strategy should help guide the design of future chemistries and find widespread use for different in vivo bioorthogonal applications, particularly in the biomedical sciences.in vivo chemistry | pharmacokinetics | PET imaging | intravital microscopy | pretargeting T he ability to perform selective chemistries in living systems such as single cells, 3D cultures, invertebrates, or mammals would have far reaching applications in tracking biomolecules, designing new therapeutic approaches, and in visualizing medically relevant biomarkers. To date, only a few practical bioorthogonal reactions have been reported, the most popular being the Staudinger ligation and the [3 þ 2] cycloaddition "click" reaction between azides and alkynes (1, 2). The latter click reaction involves copper(I) catalyzed coupling of an azide and terminal alkyne to generate a stable triazole (2). Until recently, the necessity of the copper catalyst precluded use of this reaction in biological systems due to toxicity concerns (3, 4). Bertozzi and others elegantly solved this problem by developing several new ring strained dienophile derivatives that do not require catalysts (5-9). However, many of these derivatives have low water solubility, require complex multistep synthesis, and possess suboptimal kinetics. Our search for alternative rapid, selective, and chemically accessible coupling reactions without need for a catalyst led us and others to investigate the [4 þ 2] inverse Diels-Alder cycloaddition (10-13). We realized that this set of chemistries is more uniquely suited to biological applications and may indeed represent a universal platform technology ( Fig. 1 A and B). Specifically we and others have shown that the cycloaddition between tetrazine (Tz) and trans-cyclooctene (TCO) can proceed orders of magnitude faster than previously studied azide and alkyne click reactions and importantly does not require the action of a catalyst. This reaction has also been adapted for single cell imaging using newer fluorogenic Tz probes (14).Although initial work focused on in vitro labeling there has been a surge of recent work applying this reaction for various in vivo applications (15-17). Despite the above progress, our results with initial TCO...

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mentioning

confidence: 87%

Reactive polymer enables efficient in vivo bioorthogonal chemistry

Devaraj

Thurber

Keliher

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

179

187

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“…Reaction rate is always a premium consideration for applications to nuclear medicine due to the low concentrations and short half-lives intrinsic to radiochemistry. 40, 43-51 Fast bioorthogonal reactivity is especially important for pretargeted imaging and radioimmunotherapy where the bioconjugation event takes place in vivo . 43-48 The development of more highly reactive TCO probes has also proven necessary for applications to intracellular labeling, 35, 52-57 as tetrazines that display the best long term in vivo stability also tend to be less reactive in Diels-Alder reactions.…”

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confidence: 99%

Conformationally strained trans-cyclooctene with improved stability and excellent reactivity in tetrazine ligation

et al. 2014

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Computation has guided the design of conformationally-strained dioxolane-fused trans-cyclooctene (d-TCO) derivatives that display excellent reactivity in the tetrazine ligation. A water soluble derivative of 3,6-dipyridyl-s-tetrazine reacts with d-TCO with a second order rate k2 366,000 (+/− 15,000) M−1s−1 at 25 °C in pure water. Furthermore, d-TCO derivatives can be prepared easily, are accessed through diastereoselective synthesis, and are typically crystalline bench-stable solids that are stable in aqueous solution, blood serum, or in the presence of thiols in buffered solution. GFP with a genetically encoded tetrazine-containing amino acid was site-specifically labelled in vivo by a d-TCO derivative. The fastest bioorthogonal reaction reported to date [k2 3,300,000 (+/− 40,000) M−1s−1 in H2O at 25 °C] is described herein with a cyclopropane-fused trans-cyclooctene. d-TCO derivatives display rates within an order of magnitude of these fastest trans-cyclooctene reagents, and also display enhanced stability and aqueous solubility.

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“…1,2 In recent years, the bioorthogonal reactions of strained alkenes with s -tetrazines have served as tools for rapid bioorthogonal labeling, with applications that extend to cell biology, 3–5 nuclear medicine 6–8 and materials science. 9–12 In particular, the bioorthogonal reactions of s -tetrazines with trans- cyclooctene (TCO) derivatives have been shown to be exceptionally rapid, with rate constants in excess of k 2 = 10 6 M −1 s −1 13,14.…”

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confidence: 99%

“…Cyclic RGD analog 4 has been used in combination with 18 F-labelled TCO 5 for PET imaging in live mice. 6 Recently, a 11 C-labeled derivative of 2 for PET imaging applications has also been described. 25 Fluorescently labeled s- tetrazines such as 6 have been used for site specific labeling in live mammalian cells with proteins that contain either norbornene– 15 or TCO–containing 26 amino acids.…”

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An efficient and mild oxidant for the synthesis of s-tetrazines

Selvaraj

Fox

2014

Tetrahedron Letters

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PhI(OAc)2 serves as a mild and effective oxidant for the synthesis of s-tetrazine derivatives— molecules of emerging significance to the field of bioorthogonal chemistry. This reagent serves as a complementary oxidant to harsher nitrous reagents. Use of PhI(OAc)2 improves the synthesis of 5-amino-di(pyridin-2-yl)-s-tetrazine, a molecule that has been broadly used for cellular imaging and nuclear medicine. The generality of PhI(OAc)2 as the oxidant for tetrazine synthesis is demonstrated for nine tetrazines in 75–98% yield.

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Tetrazine-trans-cyclooctene ligation for the rapid construction of integrin αvβ3 targeted PET tracer based on a cyclic RGD peptide

Cited by 92 publications

References 29 publications

Reactive polymer enables efficient in vivo bioorthogonal chemistry

Reactive polymer enables efficient in vivo bioorthogonal chemistry

Conformationally strained trans-cyclooctene with improved stability and excellent reactivity in tetrazine ligation

An efficient and mild oxidant for the synthesis of s-tetrazines

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