- Cyanine compound containing tetrazine unit and preparation method and application thereof
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The invention provides a cyanine compound containing a tetrazine unit, a preparation method of the cyanine compound and application of the cyanine compound to near-infrared fluorescence labeling of tumor cells by utilizing a biological orthogonal reaction
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- A kit-based aluminium-[18F]fluoride approach to radiolabelled microbubbles
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The production of18F-labelled microbubbles (MBs)viathe aluminium-[18F]fluoride ([18F]AlF) radiolabelling method and facile inverse-electron-demand Diels-Alder (IEDDA) ‘click’ chemistry is reported. An [18F]AlF-NODA-labelled tetrazine was synthesised in excellent radiochemical yield (>95% RCY) and efficiently conjugated to atrans-cyclooctene (TCO) functionalised phospholipid (40-50% RCY), which was incorporated into MBs (40-50% RCY). To demonstrate the potential of producing18F-labelled MBs for clinical studies, we also describe a kit-based approach which is amenable for use in a hospital radiopharmacy setting.
- Aboagye, Eric O.,Allott, Louis,Barnes, Chris,Braga, Marta,Hernández-Gil, Javier,Long, Nicholas J.,Tang, Meng-Xing,Teh, Jin Hui
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supporting information
p. 11677 - 11680
(2021/11/12)
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- Detecting hypoxia: In vitro using 18F-pretargeted IEDDA click chemistry in live cells
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We have exemplified a pretargeted approach to interrogate hypoxia in live cells using radioactive bioorthogonal inverse electron demand Diels-Alder (IEDDA) click chemistry. Our novel 18F-tetrazine probe ([18F]FB-Tz) and 2-nitroimidazole-based TCO targeting molecule (8) showed statistically significant (P a 60 min incubation of [18F]FB-Tz. This is the first time that an intracellularly targeted small-molecule for IEDDA click has been used in conjunction with a radioactive reporter molecule in live cells and may be a useful tool with far-reaching applicability for a variety of applications. This journal is
- Aboagye, Eric O.,Allott, Louis,Barnes, Chris,Braga, Marta,Brickute, Diana,Carroll, Laurence,Chen, Cen,Leung, Sau Fung Jacob,Wang, Ning
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p. 20335 - 20341
(2021/06/28)
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- New 18F-labeled dienophiles and 18F-labeling method using IeDDA reaction with tetrazines
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The present invention relates to a method of labeling of F-18 radioisotopes using an inverse electron demand Diels-Alder (IeDDA) reaction between a tetrazine compound and a dienophile. A ketone compound according to the present invention easily forms an enamine with a secondary amine in an aqueous solution phase to cause significantly rapid conjugation with a tetrazine compound. Particularly, as the amount of water is increased in the aqueous solution, the reaction proceeds well, so that the compound may be suitable for a bioactive compound, such as a peptide or antibody. Unlike conventional reactions producing different stereoisomers, the reaction produces a small amount of only one type of stereoisomer, besides a main product, and the product can be purified through high performance liquid chromatography, etc. Further, as compared to the conventional ^18F-labeled tetrazine or ^18F-labeled TCO synthesis with a low yield, the ^18F-labeled ketone compound represented by the following chemical formula 1 can be obtained with a high ^18F-labeling yield from a precursor thereof.COPYRIGHT KIPO 2021
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- Inverse Electron-Demand Diels-Alder Bioconjugation Reactions Using 7-Oxanorbornenes as Dienophiles
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Oligonucleotides, peptides, and peptide nucleic acids incorporating 7-oxanorbornene as a dienophile were reacted with tetrazines linked to either a peptide, d-biotin, BODIPY, or N-acetyl-d-galactosamine. The inverse electron-demand Diels-Alder (IEDDA) cycloaddition, which was performed overnight at 37 °C, in all cases furnished the target conjugate in good yields. IEDDA reactions with 7-oxanorbornenes produce a lower number of stereoisomers than that of IEDDA cycloadditions with other dienophiles.
- Agramunt, Jordi,Ginesi, Rebecca,Grandas, Anna,Pedroso, Enrique
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p. 6593 - 6604
(2020/07/14)
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- Bioorthogonal "labeling after Recognition" Affording an FRET-Based Luminescent Probe for Detecting and Imaging Caspase-3 via Photoluminescence Lifetime Imaging
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Bis-labeling with a luminescent energy donor/acceptor pair onto biological substrates affords probes which give FRET readouts for the detection of interaction partners. However, the covalently bound luminophores bring about steric hindrance and nonspecific interaction, which probably perturb the biological recognition. Herein, we designed a highly sensitive and specific "labeling after recognition" sensing approach, where luminophore labeling occurred after the biological recognition. Taking the cutting enzyme caspase-3 as an example, we demonstrated the detection of its catalytic activity in solution and apoptotic cells using the tetrapeptide motif Asp-Glu-Val-Asp (DEVD) as the cleavable substrate, and an iridium(III) complex and a rhodamine derivative as the energy donor/acceptor pair. The DEVD tetrapeptide was modified with an azide and a GK-norbornylene groups at the amino and carboxyl terminuses, respectively, which allowed donor/acceptor bis-labeling via two independent catalysis-free bioorthogonal reactions. The phosphorescence lifetime of the iridium(III) complex was quenched upon bis-labeling owing to the intracellular FRET to the rhodamine derivative, and significantly elongated upon the peptide being catalytically cleaved by caspase-3. Interestingly, the sensitivity and efficiency of the lifetime responses were much higher in the "labeling after recognition" sensing approach. Molecular docking analysis showed that the steric hindrance and nonspecific interactions partially inhibited the biological recognition of the DEVD substrate by caspase-3. The imaging of the catalytic activity of caspase-3 in apoptotic cells was demonstrated via photoluminescence lifetime imaging microscopy. Lifetime analysis not only confirmed the occurrence of intracellular bioorthogonal bis-labeling and catalytic cleavage, but also showed the extent to which the two dynamic processes occurred.
- Dai, Peiling,Huang, Wei,Liu, Shujuan,Song, Linna,Wang, Ling,Wang, Yun,Wu, Qi,Zhang, Kenneth Yin,Zhao, Qiang,Zhu, Hengyu
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supporting information
p. 1057 - 1064
(2020/02/20)
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- Development of 68Ga-labelled ultrasound microbubbles for whole-body PET imaging
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Microbubble (MB) contrast agents have revolutionalised the way ultrasound (US) imaging can be used clinically and pre-clinically. Contrast-enhanced US offers improvements in soft-tissue contrast, as well as the ability to visualise disease processes at th
- Hernández-Gil, Javier,Braga, Marta,Harriss, Bethany I.,Carroll, Laurence S.,Leow, Chee Hau,Tang, Meng-Xing,Aboagye, Eric O.,Long, Nicholas J.
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p. 5603 - 5615
(2019/06/08)
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- Installation of Minimal Tetrazines through Silver-Mediated Liebeskind-Srogl Coupling with Arylboronic Acids
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Described is a general method for the installation of a minimal 6-methyltetrazin-3-yl group via the first example of a Ag-mediated Liebeskind-Srogl cross-coupling. The attachment of bioorthogonal tetrazines on complex molecules typically relies on linkers that can negatively impact the physiochemical properties of conjugates. Cross-coupling with arylboronic acids and a new reagent, 3-((p-biphenyl-4-ylmethyl)thio)-6-methyltetrazine (b-Tz), proceeds under mild, PdCl2(dppf)-catalyzed conditions to introduce minimal, linker-free tetrazine functionality. Safety considerations guided our design of b-Tz which can be prepared on decagram scale without handling hydrazine and without forming volatile, high-nitrogen tetrazine byproducts. Replacing conventional Cu(I) salts used in Liebeskind-Srogl cross-coupling with a Ag2O mediator resulted in higher yields across a broad library of aryl and heteroaryl boronic acids and provides improved access to a fluorogenic tetrazine-BODIPY conjugate. A covalent probe for MAGL incorporating 6-methyltetrazinyl functionality was synthesized in high yield and labeled endogenous MAGL in live cells. This new Ag-mediated cross-coupling method using b-Tz is anticipated to find additional applications for directly introducing the tetrazine subunit to complex substrates.
- Lambert, William D.,Fang, Yinzhi,Mahapatra, Subham,Huang, Zhen,Am Ende, Christopher W.,Fox, Joseph M.
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supporting information
p. 17068 - 17074
(2019/11/16)
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- Fast and pH-Independent Elimination of trans-Cyclooctene by Using Aminoethyl-Functionalized Tetrazines
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The inverse-electron-demand Diels–Alder/pyridazine elimination tandem reaction, in which the allylic substituent on trans-cyclooctene is eliminated following reaction with tetrazines, is gaining interest as a versatile bioorthogonal process. One potential shortcoming of such currently used reactions is their propensity to proceed faster and more efficiently at lower pH, a feature caused by the nature of the tetrazines used. Here, we present aminoethyl-substituted tetrazines as the first pH-independent reagents showing invariably fast elimination kinetics at all biologically relevant pH values.
- Sarris, Alexi J. C.,Hansen, Thomas,de Geus, Mark A. R.,Maurits, Elmer,Doelman, Ward,Overkleeft, Herman S.,Codée, Jeroen D. C.,Filippov, Dmitri V.,van Kasteren, Sander I.
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p. 18075 - 18081
(2018/11/23)
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- LABELING OF ANTIBODIES
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Provided herein are methods for producing site specific PEG modifications to single domain antibodies (e.g., VHHs). Methods for producing site- specific ally conjugated bivalent single domain antibodies (e.g., VHHs) are also provided. Methods for labeling
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- Enzyme-Mediated Modification of Single-Domain Antibodies for Imaging Modalities with Different Characteristics
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Antibodies are currently the fastest-growing class of therapeutics. Although naked antibodies have proven valuable as pharmaceutical agents, they have some limitations, such as low tissue penetration and a long circulatory half-life. They have been conjugated to toxic payloads, PEGs, or radioisotopes to increase and optimize their therapeutic efficacy. Although nonspecific conjugation is suitable for most in vitro applications, it has become evident that site specifically modified antibodies may have advantages for in vivo applications. Herein we describe a novel approach in which the antibody fragment is tagged with two handles: One for the introduction of a fluorophore or 18F isotope, and the second for further modification of the fragment with a PEG moiety or a second antibody fragment to tune its circulatory half-life or its avidity. Such constructs, which recognize Class II MHC products and CD11b, showed high avidity and specificity. They were used to image cancers and could detect small tumors.
- Rashidian, Mohammad,Wang, Lu,Edens, Jerre G.,Jacobsen, Johanne T.,Hossain, Intekhab,Wang, Qifan,Victora, Gabriel D.,Vasdev, Neil,Ploegh, Hidde,Liang, Steven H.
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p. 528 - 533
(2016/02/27)
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- Rigid tetrazine fluorophore conjugates with fluorogenic properties in the inverse electron demand Diels-Alder reaction
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1,2,4,5-Tetrazine fluorophore derivatives with structurally rigid molecular designs were synthesized using Sonogashira and Stille cross-coupling as well as copper-catalyzed azide-alkyne cycloaddition. The synthesized bichromophoric systems exhibit low flu
- Wieczorek, Achim,Buckup, Tiago,Wombacher, Richard
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supporting information
p. 4177 - 4185
(2014/06/10)
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- A bioorthogonal 68Ga-labelling strategy for rapid in vivo imaging
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Herein, we describe a fast and robust method for achieving 68Ga-labelling of the EGFR-selective monoclonal antibody (mAb) Cetuximab using the bioorthogonal Inverse-electron-Demand Diels-Alder (IeDDA) reaction. The in vivo imaging of EGFR is demonstrated, as well as the translation of the method within a two-step pretargeting strategy. This journal is the Partner Organisations 2014.
- Evans, Helen L.,Nguyen, Quang-De,Carroll, Laurence S.,Kaliszczak, Maciej,Twyman, Frazer J.,Spivey, Alan C.,Aboagye, Eric O.
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p. 9557 - 9560
(2014/08/18)
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- NOVEL TETRAZINES AND METHOD OF SYNTHESIZING THE SAME
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Provided herein, inter alia, are compositions and methods of synthesis and detection of tetrazines and diazonorcaradienes.
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Paragraph 0285
(2013/10/22)
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- Metal-catalyzed one-pot synthesis of tetrazines directly from aliphatic nitriles and hydrazine
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Paving the way: The lack of convenient synthetic methods is a significant roadblock to the broader use of 1,2,4,5-tetrazines in bioorthogonal chemistry and functional materials. Lewis acid metal catalysts-most notably divalent nickel and zinc salts-are described to catalyze the one-pot synthesis of 1,2,4,5-tetrazines directly from aliphatic nitriles (see scheme). Copyright
- Yang, Jun,Karver, Mark R.,Li, Weilong,Sahu, Swagat,Devaraj, Neal K.
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supporting information; experimental part
p. 5222 - 5225
(2012/07/14)
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