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Biology

Микроволновая при содействии одного банка Синтез N-Сукцинимидил-4-[ 18 F] fluorobenzoate ([ 18 F] SFB)

Published: June 28, 2011
doi:
10.3791/2755
, , , , ,
1Department of Molecular and Medical Pharmacology, David Geffen School of Medicine,University of California at Los Angeles, 2Crump Institute for Molecular Imaging, David Geffen School of Medicine,University of California at Los Angeles, 3California NanoSystems Institute,University of California at Los Angeles, 4Nuclear Medicine, PET Center, Shanghai Medical Collegea,Fudan University, 5Electronics and Information Engineering, College of Electronics and Information Engineering,Wuhan Textile University

Summary

Поверхностное, с одним горшком синтез N-сукцинимидил-4-[<sup> 18</sup> F] fluorobenzoate ([<sup> 18</sup> F] SFB) был разработан на основе неводных, трехступенчатый радиохимического процесса. Использование микроволнового нагрева, вся процедура может быть завершена менее чем за 30 мин, или 60 мин с последующей очистки с помощью препаративной ВЭЖХ. Распад коррекцией радиохимического дает (RCYs) были 35-5% (п> 30).

Abstract

Biomolecules, including peptides,1-9 proteins,10,11 and antibodies and their engineered fragments,12-14 are gaining importance as both potential therapeutics and molecular imaging agents. Notably, when labeled with positron-emitting radioisotopes (e.g., Cu-64, Ga-68, or F-18), they can be used as probes for targeted imaging of many physiological and pathological processes.15-18 Therefore, significant effort has devoted to the synthesis and exploration of 18F-labeled biomolecules. Although there are elegant examples of the direct 18F-labeling of peptides,19-22 the harsh reaction conditions (i.e., organic solvent, extreme pH, high temperature) associated with direct radiofluorination are usually incompatible with fragile protein samples. To date, therefore, the incorporation of radiolabeled prosthetic groups into biomolecules remains the method of choice.23,24

N-Succinimidyl-4-[18F]fluorobenzoate ([18F]SFB),25-37 a Bolton-Hunter type reagent that reacts with the primary amino groups of biomolecules, is a very versatile prosthetic group for the 18F-labeling of a wide spectrum of biological entities, in terms of its evident in vivo stability and high radiolabeling yield. After labeling with [18F]SFB, the resulting [18F]fluorobenzoylated biomolecules could be explored as potential PET tracers for in vivo imaging studies.1 Most [18F]SFB radiosyntheses described in the current literatures require two or even three reactors and multiple purifications by using either solid phase extraction (SPE) or high-performance liquid chromatography (HPLC). Such lengthy processes hamper its routine production and widespread applications in the radiolabeling of biomolecules. Although several module-assisted [18F]SFB syntheses have been reported,29-32, 41-42 they are mainly based on complicated and lengthy procedures using costly commercially-available radiochemistry boxes (Table 1). Therefore, further simplification of the radiosynthesis of [18F]SFB using a low-cost setup would be very beneficial for its adaption to an automated process.

Herein, we report a concise preparation of [18F]SFB, based on a simplified one-pot microwave-assisted synthesis (Figure 1). Our approach does not require purification between steps or any aqueous reagents. In addition, microwave irradiation, which has been used in the syntheses of several PET tracers,38-41 can gives higher RCYs and better selectivity than the corresponding thermal reactions or they provide similar yields in shorter reaction times.38 Most importantly, when labeling biomolecules, the time saved could be diverted to subsequent bioconjugation or PET imaging step.28,43 The novelty of our improved [18F]SFB synthesis is two-fold: (1) the anhydrous deprotection strategy requires no purification of intermediate(s) between each step and (2) the microwave-assisted radiochemical transformations enable the rapid, reliable production of [18F]SFB.

Protocol

1. Первоначальная подготовка V-флаконе (5-мл) RV1 (с мешалкой) используется как основной реактор для выполнения микроволновой синтез. Это связано с PEEK адаптер с семью входа / выхода порта соединяется и размещены внутри микроволновой полости (см. Рисунок 2). RV2 подключен к SPE картридж (Я) д…

Discussion

Эта упрощенная три шага, с одним горшком radiosynthesis из 18 F-ацилирования реагента [18 F] SFB разработан на основе неводных химии. Этот процесс имеет превосходную воспроизводимость и могут быть использованы для надежного производства [18 F] SFB в автоматизированных модулей радиох…

Disclosures

The authors have nothing to disclose.

Acknowledgements

Это исследование было поддержано Министерством энергетики США (DE-FG02-09ER09-08 и DE-PS02-09ER09-18), рака Йонссон всеобъемлющем центр в Лос-Анджелесе, и в отрасли университета Совместная исследовательская программа (UC Discovery Грант, bio07 -10 665). Мы благодарим доктора Nagichettiar Satyamurthy и штабов на установке UCLA биомедицинских Циклотрон для обеспечения F-18 радиоизотопных и много дискуссий проницательными. Мы благодарим доктора. Майкл Коллинз, Грег Леблан, Джозеф Ламберт, и Келлер Барнхардт от CEM за технические консультации и поддержку. Мы благодарим Дирк Уильямс, Уильямс Дарин, д-ра. Иосиф Hong Дун Лин, и Майкл Ван Дам для проектирования и обработки деталей для изменения реактора CEM микроволновой печью и для SPE модули очистки.

Materials

Name of the reagent Company Catalogue number Comments (optional)
acetic acid in aqueous solution (5%, v/v) Fisher A38-500 Prepared in our lab
Acetonitrile Sigma-Aldrich 75-05-8  
Diethyl ether Sigma-Aldrich 14775  
Dimethyl Sulfoxide (DMSO) Sigma-Aldrich 472301  
Ethyl 4-(N,N,N-trimethylammonium)benzoate triflate Prepared in lab    
4,7,13,16,21,24-Hexaoxa-1,10-diazabicyclo[8.8.8]hexacosane (K222) Sigma-Aldrich 29,111-0  
O-(N-succinimidyl)-N,N,N‘,N‘-tetramethyluronium tetrafluoroborate (TSTU) Sigma-Aldrich 105832-38-0  
Potassium carbonate in aqueous solution (1M) Sigma-Aldrich 209619 Prepared in our lab
Potassium tert-butoxide Sigma-Aldrich 156671  
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Tags

Microwave-assistedOne-pot SynthesisN-succinimidyl-4-[18F]fluorobenzoate[18F]SFBBiomoleculesPeptidesProteinsAntibodiesEngineered FragmentsTherapeuticsMolecular Imaging AgentsPositron-emitting RadioisotopesCu-64Ga-68F-18Targeted ImagingPhysiological ProcessesPathological ProcessesSynthesisExploration18F-labeled BiomoleculesDirect 18F-labelingOrganic SolventExtreme PHHigh TemperatureRadiolabeled Prosthetic GroupsMethod Of ChoiceBolton-Hunter Type ReagentPrimary Amino GroupsIn Vivo StabilityRadiolabeling YieldPET Tracers

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Hou, S., Phung, D. L., Lin, W., Wang, M., Liu, K., Shen, C. K. Microwave-assisted One-pot Synthesis of N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB). J. Vis. Exp. (52), e2755, doi:10.3791/2755 (2011).
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