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Synthesis of high-molar-activity [18F]6-fluoro-l-DOPA suitable for human use via Cu-mediated fluorination of a BPin precursor

Nature Protocols volume 15, pages 1742–1759 (2020)Cite this article

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Abstract

[18F]6-fluoro-l-DOPA ([18F]FDOPA) is a diagnostic radiopharmaceutical for positron emission tomography (PET) imaging that is used to image Parkinson’s disease, brain tumors, and focal hyperinsulinism of infancy. Despite these important applications, [18F]FDOPA PET remains underutilized because of synthetic challenges associated with accessing the radiotracer for clinical use; these stem from the need to radiofluorinate a highly electron-rich catechol ring in the presence of an amino acid. To address this longstanding challenge in the PET radiochemistry community, we have developed a one-pot, two-step synthesis of high-molar-activity [18F]FDOPA by Cu-mediated fluorination of a pinacol boronate (BPin) precursor. The method is fully automated, has been validated to work well at two separate sites (an academic facility with a cyclotron on site and an industry lab purchasing [18F]fluoride from an outside vendor), and provides [18F]FDOPA in reasonable radiochemical yield (2.44 ± 0.70 GBq, 66 ± 19 mCi, 5 ± 1%), excellent radiochemical purity (>98%) and high molar activity (76 ± 30 TBq/mmol, 2,050 ± 804 Ci/mmol), n = 26. Herein we report a detailed protocol for the synthesis of [18F]FDOPA that has been successfully implemented at two sites and validated for production of the radiotracer for human use.

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Fig. 1: Radiosyntheses of [18F]FDOPA and motivation for this work.
Fig. 2: Radiosyntheses of [18F]FDOPA and the TRACERLab automated synthesis module.
Fig. 3: Semi-preparative HPLC traces for [18F]FDOPA prepared using two different methods.
Fig. 4: Analytical HPLC traces of [18F]FDOPA using a Luna NH2 analytical column.
Fig. 5: Chiral HPLC trace of production of [18F]FDOPA, 6F-d,l-DOPA reference standard, and [6F]-l-DOPA reference standard using a Chirobiotic T analytical column.

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All data generated or analyzed during this study are included in this published article (and its Supplementary Information files).

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Acknowledgements

We acknowledge the NIH (R01EB021155 to M.S.S. and P.J.H.S.) and US DOE/NIBIB (DE-SC0012484 to P.J.H.S.) for financial support.

Author information

Author notes

  1. Andrew V. Mossine

    Present address: Curium Pharma, Nuclear Medicine Manufacturing, Noblesville, IN, USA

  2. Katarina J. Makaravage

    Present address: Gordon Center for Medical Imaging, Massachusetts General Hospital, Harvard Medical School, Cambridge, MA, USA

  3. Naoko Ichiishi

    Present address: Takeda Pharmaceuticals International Co., Process Chemistry, Boston, MA, USA

  4. Jason M. Miller

    Present address: Environmental Analysis Branch, US Army Corps of Engineers, Detroit, MI, USA

Authors and Affiliations

  1. Department of Radiology, University of Michigan, Ann Arbor, MI, USA

    Andrew V. Mossine, Allen F. Brooks, Bradford D. Henderson & Peter J. H. Scott

  2. Department of Medicinal Chemistry, University of Michigan, Ann Arbor, MI, USA

    Sean S. Tanzey, Jason M. Miller & Peter J. H. Scott

  3. Department of Chemistry, University of Michigan, Ann Arbor, MI, USA

    Katarina J. Makaravage, Naoko Ichiishi & Melanie S. Sanford

  4. AbbVie Deustschland GmbH & Co. KG Ludwigschafen, Ludwigshafen, Germany

    Thomas Erhard & Christian Bruetting

  5. AbbVie Translational Imaging, North Chicago, IL, USA

    Marc B. Skaddan

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Contributions

A.V.M., A.F.B., N.I., P.J.H.S., and M.S.S. conceived and developed the original radiofluorination of organoborons used herein to prepare [18F]FDOPA. T.E. and C.B. synthesized BPin precursor 1. A.V.M., S.S.T., A.F.B., K.J.M., N.I., J.M.M., and M.B.S. performed radiofluorination reactions. B.D.H. and M.B.S. performed QC testing. M.S.S. and P.J.H.S. provided supervision and funding. All authors analyzed data and participated in the writing and editing of the manuscript.

Corresponding authors

Correspondence to Melanie S. Sanford or Peter J. H. Scott.

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Peer review information Nature Protocols thanks Bernd Neumaier and the other, anonymous, reviewer(s) for their contribution to the peer review of this work

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Related links

Key reference using this protocol

Mossine A. V. et al. Org. Biomol. Chem. 17, 8701–8705 (2019): https://doi.org/10.1039/C9OB01758E

Key data used in this protocol

Mossine A. V. et al. Org. Biomol. Chem. 17, 8701–8705 (2019): https://doi.org/10.1039/C9OB01758E

Integrated supplementary information

Supplementary Figure 1

Standard TRACERLab FXFN Configuration for One-pot Synthesis of [18F]FDOPA (reproduced with permission of GE Healthcare).

Supplementary Figure 2

Modified TRACERLab FXFN Configuration for alternative synthesis of [18F]FDOPA with HLB purification between fluorination and deprotection (Adapted from Supplementary Fig. 1 and reproduced with permission of GE Healthcare).

Supplementary Figure 3 Analytical trace (RAD top, 282 nm UV bottom) of [18F]FDOPA at end-of-synthesis.

Column: Luna NH2 5 micron 4.6x150 mm column; mobile phase: 70% MeCN 10 mM KOAc, pH 5.2; flow rate: 1.5 mL/min. [18F]FDOPA prepared using alternative synthesis with HLB purification between fluorination and deprotection.

Supplementary Figure 4 Analytical trace (RAD top, 282 nm UV bottom) of [18F]FDOPA 4 h post-end-of-synthesis.

Column: Luna NH2 5 micron 4.6 × 150 mm column; mobile phase: 70% MeCN 10 mM KOAc, pH 5.2; flow rate: 1.5 mL/min. [18F]FDOPA prepared using alternative synthesis with HLB purification between fluorination and deprotection.

Supplementary information

Supplementary Information

Supplementary Figs. 1–4 and Supplementary Methods 1 and 2.

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Mossine, A.V., Tanzey, S.S., Brooks, A.F. et al. Synthesis of high-molar-activity [18F]6-fluoro-l-DOPA suitable for human use via Cu-mediated fluorination of a BPin precursor. Nat Protoc 15, 1742–1759 (2020). https://doi.org/10.1038/s41596-020-0305-9

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