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Short and scalable synthesis of an anhydride precursor of the environment-sensitive fluorophore 6-dimethylaminonaphthalimide

Nature Protocols volume 6, pages 1990–1997 (2011)Cite this article

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Abstract

Environment-sensitive fluorophores have different quantum yields in different solvents. 6-Dimethylaminonaphthalimide (6-DMN), for example, has a low quantum yield in aqueous solutions, but is highly fluorescent in nonpolar solvents or when bound to hydrophobic sites in proteins or membranes. 6-DMN has been used to investigate protein-protein interactions, as well as the in-tissue distribution and internalization of δ-receptors. This protocol describes a highly efficient three-stage synthesis of 6-dimethylamino-2,3-naphthalenedicarboxylic anhydride (compound 1), which is a stable precursor that can be converted to 6-DMN. The three stages are (i) photo-bromination of 4-nitro-o-xylene (yield 82%), (ii) Diels-Alder reaction followed by base hydrolysis, resulting in 6-nitro-2,3-naphthalenedicarboxylic acid (yield 87%) and (iii) reductive amination followed by dehydration (yield 76.5%) to form compound 1. The synthesis can be performed on a gram scale (in 55 h over 3 d) with an overall yield of about 55%. It can easily be modified to prepare related compounds by, for example, performing different reactions using 6-nitro-2,3-naphthalenedicarboxylic acid.

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Figure 1: 6-Dimethylamino-2,3-naphthalimide (6-DMN) derivatives and applications.
Figure 2: General approach to the synthesis of 6-substituted 2,3-naphthalimide analogs.
Figure 3: Previous synthesis.
Figure 4: Small- and large-scale preparation of 6-DMN precursor.
Figure 5: Equipment setup and appearance of the photo-bromination reaction.

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Acknowledgements

This work is supported by the National Basic Research Program of China (973 Program, 2010CB126100), the National High Technology Research and Development Program of China (863 Program 2011AA10A207), the Shanghai Leading Academic Discipline Project (B507) and the Fundamental Research Funds for the Central Universities. We thank Shanghai Municipal Government (SMGS) for funding and J. Srivani for valuable support.

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Authors and Affiliations

  1. Shanghai Key Laboratory of Chemical Biology,

    Kishore Baathulaa, Yufang Xu & Xuhong Qian

  2. , School of Pharmacy, East China University of Science and Technology, Shanghai, China

    Kishore Baathulaa, Yufang Xu & Xuhong Qian

Authors
  1. Kishore Baathulaa
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Contributions

K.B. conducted the synthesis and determined the synthetic routes, and prepared the first draft and final version of the manuscript. X.Q. supervised all of the work and is the corresponding author. Y.X. prepared the final version of the manuscript and provided senior authorship.

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Correspondence to Xuhong Qian.

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Baathulaa, K., Xu, Y. & Qian, X. Short and scalable synthesis of an anhydride precursor of the environment-sensitive fluorophore 6-dimethylaminonaphthalimide. Nat Protoc 6, 1990–1997 (2011). https://doi.org/10.1038/nprot.2011.415

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