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Biology

माइक्रोवेव की मदद से एक पॉट संश्लेषण एन Succinimidyl-4-[ एफ] ([ फा] 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

एक सतही, एक पॉट संश्लेषण एन succinimidyl चार – [<sup18></supFluorobenzoate> एफ] ([<sup18></sup> फा] SFB) एक गैर जलीय radiochemical तीन कदम प्रक्रिया के आधार पर विकसित किया गया था. माइक्रोवेव हीटिंग का प्रयोग, पूरी प्रक्रिया में कम से कम 30 मिनट, या प्रारंभिक HPLC द्वारा आगे शुद्धि के साथ 60 मिनट में पूरा किया जा सकता है. क्षय सही radiochemical पैदावार (RCYs) 35-5% (n> 30) थे.

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. प्रारंभिक तैयारी (5 एमएल) वि शीशी RV1 बार सरगर्मी के साथ माइक्रोवेव संश्लेषण प्रदर्शन करने के लिए मुख्य प्रतिक्रिया पोत के रूप में प्रयोग किया जाता है. यह सात / इनलेट आउटलेट बंदरगाहों के साथ एक तिरछी ?…

Discussion

यह सरल तीन कदम, एक बर्तन 18 एफ acylation अभिकर्मक के radiosynthesis [18 एफ] SFB गैर जलीय रसायन शास्त्र पर आधारित विकसित की है. इस प्रक्रिया को उत्कृष्ट reproducibility है और मज़बूती के उत्पादन के लिए इस्तेमाल किया जा सकता [18 …

Disclosures

The authors have nothing to disclose.

Acknowledgements

इस अध्ययन अमेरिकी ऊर्जा विभाग (de-FG02-09ER09-08 और de-PS02 – 09ER09 18), UCLA में Jonsson व्यापक कैंसर केंद्र, और उद्योग – विश्वविद्यालय सहकारी रिसर्च प्रोग्राम (यूसी डिस्कवरी अनुदान, bio07 द्वारा समर्थित किया गया -10,665). हम UCLA बायोमेडिकल साइक्लोट्रॉन सुविधा डा. Nagichettiar सत्यमूर्ति और कर्मचारी F-18 रेडियो आइसोटोप और कई व्यावहारिक चर्चा प्रदान करने के लिए धन्यवाद. हम डीआरएस धन्यवाद. माइकल कोलिन्स, ग्रेग Leblanc, यूसुफ Lambert, और उनकी तकनीकी सलाह और समर्थन के लिए CEM से केलर Barnhardt. हम एक प्रकार की कटार विलियम्स, डरिन विलियम्स, डीआरएस धन्यवाद. यूसुफ हांग डुन लिन, और माइकल डिजाइन और मशीनिंग भागों के लिए वैन बांध CEM माइक्रोवेव रिएक्टर को संशोधित करने के लिए और विशेष शोधन मॉड्यूल के लिए.

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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