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Click processes orthogonal to CuAAC and SuFEx forge selectively modifiable fluorescent linkers

Nature Chemistry volume 16, pages 426–436 (2024)Cite this article

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Abstract

The appeal of catalytic click chemistry is largely due to the copper-catalysed azide–alkyne cycloaddition (CuAAC) process, which is orthogonal to the more recently introduced sulfur–fluoride exchange (SuFEx). However, the triazole rings generated by CuAAC are not readily modifiable, and SuFEx connectors cannot be selectively functionalized, attributes that would be attractive in a click process. Here we introduce bisphosphine–copper-catalysed phenoxydiazaborinine formation (CuPDF), a link-and-in situ modify strategy for merging a nitrile, an allene, a diborane and a hydrazine. We also present copper- and palladium-catalysed quinoline formation (Cu/PdQNF), which is applicable in aqueous media, involving an aniline as the modifier. CuPDF and Cu/PdQNF are easy to perform and deliver robust, alterable and tunable fluorescent hubs. CuPDF and Cu/PdQNF are orthogonal to SuFEx and CuAAC, despite the latter and CuPDF also being catalysed by an organocopper species. These advantages were applied to protecting group-free syntheses of sequence-defined branched oligomers, a chemoselectively amendable polymer, three drug conjugates and a two-drug conjugate.

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Fig. 1: State of the art in catalytic click chemistry, and the potential impact of a link-and-in situ modify click strategy that is orthogonal to CuAAC and SuFEx.
Fig. 2: A link-and-in situ modify click process (CuPDF) that is copper-catalysed yet orthogonal to CuAAC.
Fig. 3: Efficient synthesis of uniform, sequence-defined branched oligomers by combining CuPDF, CuAAC and SuFEx.
Fig. 4: Efficient synthesis of uniform, sequence-defined branched oligomers by combining CuPDF, CuAAC and SuFEx.
Fig. 5: Protecting group-free synthesis of a polymer.
Fig. 6: Efficient bioconjugation/fluorescent marking.

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Data availability

All data in support of the findings of this study are available within the Article and its Supplementary Information.

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Acknowledgements

Funding was provided by the ANR (project PRACTACAL), CNRS and the Jean-Marie Lehn Research Foundation at the University of Strasbourg. The early stages of this work were supported by the National Institutes of Health (GM-130395). K.E.L. was supported by a Complex Systems Chemistry (CSC) graduate fellowship funded by the French National Research Agency (CSC-IGS ANR-17-EURE-0016). We thank S.-Y. Liu, J. Niu and A. Chatterjee for helpful discussions.

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Authors and Affiliations

  1. Supramolecular Science and Engineering Institute, University of Strasbourg, Strasbourg, France

    Paulo H. S. Paioti, Katherine E. Lounsbury, Filippo Romiti, Michele Formica, Valentin Bauer, Claudio Zandonella & Amir H. Hoveyda

  2. Department of Chemistry, Merkert Chemistry Center, Boston College, Chestnut Hill, MA, USA

    Michele Formica, Meagan E. Hackey, Juan del Pozo & Amir H. Hoveyda

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  1. Paulo H. S. Paioti
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Contributions

P.H.S.P., J.d.P., F.R. and A.H.H. conceived the concept. P.H.S.P., M.F. and C.Z. designed and performed the studies regarding the synthesis of sequence-defined oligomers and polymers, and K.E.L., F.R. and V.B. planned and carried out the bioconjugation studies. J.d.P. and M.E.H. investigated the mechanistic aspects of the CuPDF process. The investigations were directed by A.H.H., who composed the manuscript with revisions provided by the other authors.

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Correspondence to Amir H. Hoveyda.

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Nature Chemistry thanks Shengtao Deng, Andre Isaacs, Christopher Smedley and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 Merger of CuPDF, CuAAC and SuFEx for synthesis of a non-uniform, sequence-defined polymer.

As an example, subjection of two-pronged monomers iii and iv on the one hand, and v and vi on the other, to the envisioned link-and-in situ modify conditions would deliver complementary dimers VII and IX (via VI and VIII, respectively). While VII would contain a fluorosulfate and a terminal alkyne, IX would be equipped with an azide (for CuAAC, in red) and a silyl ether (for SuFEx, in brown). Simultaneous subjection of an equal mixture of VII and IX to copper and base catalysts needed for CuAAC and SuFEx, respectively, would afford sequence-defined polymer X with amendable and differentiable linkers, depending on the amino alcohols involved in the modify steps (namely, G1 and G2). Hence, after just four chemical steps, performed in three vessels, and without the need for protection/deprotection or any other type of functional group adjustments, four monomers iii-vi could be transformed to polymer X. CuAAC, Cu-catalysed azide–alkyne cycloaddition; SuFEx, sulfur-fluoride exchange; M, monomer.

Extended Data Fig. 2 Insight regarding the orthogonality of CuPDF and CuAAC and the uniqueness of the phos ligand.

a, CuAAC (in red) with phos–CuB(pin) is considerably less effective than the more established protocols (for example, condition A, Fig. 2c). b, Experimental evidence indicating that addition of phos–CuB(pin) to an allene is faster than an alkyne. c, With dppf as the ligand, the desired three-component process affording 6 is less efficient than when phos is used (compare to data in Fig. 2b). d, Additionally, with dppf as the ligand, chemoselectivity is lower than when phos is used: addition of the corresponding Cu–B(pin) complex to alkyne 2 is more competitive. Ts, p-toluenesulfonate; pin, pinacolato; dppf, 1,1’-bis(diphenylphosphino)ferrocene; CuAAC, copper(I)-catalysed alkyne-azide cycloaddition; Bz, benzoate.

Extended Data Fig. 3 A Cu/PdQNF process that furnishes a different linker (compared to 12a).

By altering the structure the aniline, the resulting quinoline can possess distinct physical (excitation and emission bands) and chemical attributes (clickable nitrile). Cu/PdQNF (in purple), copper- and palladium-catalysed quinoline formation. The colour of the linker corresponds to that of the fluorescent emission. Ac, acyl; Bn, benzyl; pin, pinacolato; Trt (trityl); dppf, diphenylphosphinoferrocene; TFA, trifluoroacetic acid; DTT, dithiothreitol; TFA, trifluoroacetic acid.

Supplementary information

Supplementary Information

Supplementary Figs. 1–60 and NMR spectra for all compounds.

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Paioti, P.H.S., Lounsbury, K.E., Romiti, F. et al. Click processes orthogonal to CuAAC and SuFEx forge selectively modifiable fluorescent linkers. Nat. Chem. 16, 426–436 (2024). https://doi.org/10.1038/s41557-023-01386-9

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