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Biocatalytic oxidative cross-coupling reactions for biaryl bond formation

Nature volume 603, pages 79–85 (2022)Cite this article

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Abstract

Biaryl compounds, with two connected aromatic rings, are found across medicine, materials science and asymmetric catalysis1,2. The necessity of joining arene building blocks to access these valuable compounds has inspired several approaches for biaryl bond formation and challenged chemists to develop increasingly concise and robust methods for this task3. Oxidative coupling of two C–H bonds offers an efficient strategy for the formation of a biaryl C–C bond; however, fundamental challenges remain in controlling the reactivity and selectivity for uniting a given pair of substrates4,5. Biocatalytic oxidative cross-coupling reactions have the potential to overcome limitations inherent to numerous small-molecule-mediated methods by providing a paradigm with catalyst-controlled selectivity6. Here we disclose a strategy for biocatalytic cross-coupling through oxidative C–C bond formation using cytochrome P450 enzymes. We demonstrate the ability to catalyse cross-coupling reactions on a panel of phenolic substrates using natural P450 catalysts. Moreover, we engineer a P450 to possess the desired reactivity, site selectivity and atroposelectivity by transforming a low-yielding, unselective reaction into a highly efficient and selective process. This streamlined method for constructing sterically hindered biaryl bonds provides a programmable platform for assembling molecules with catalyst-controlled reactivity and selectivity.

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Fig. 1: Biaryl bond formation through direct oxidative cross-coupling.
Fig. 2: Scope of oxidative cross-coupling reactions catalysed by wild-type P450 KtnC.
Fig. 3: Engineering P450 biocatalysts for improved activity and selectivity.
Fig. 4: Exploring alternative sequence space for catalysts with complementary reactivity.

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Data availability

Raw data associated with biocatalytic reactions and protein engineering efforts supporting the conclusions of this work are available from the corresponding author upon reasonable request.

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Acknowledgements

This research was supported by funds from the University of Michigan’s Life Sciences Institute and the Chemistry Department, Alfred P. Sloan Foundation, and Research Corporation Cottrell Scholars programme. Initial studies on the selectivity of dimerization reactions were supported by the National Institutes of Health (NIH; R35 GM124880), and protein engineering (rounds 4–7) and bioinformatic-based library generation were supported by generous funds from the Novartis Global Scholars Program. L.E.Z. is grateful to the NIH National Center for Complementary and Integrative Health (F31AT010973), J.A.Y. thanks the National Science Foundation Graduate Research Fellowship Program, A.L.L. acknowledges the Rackham Graduate School (University of Michigan) and the NIH (F31 NS111906) for funding. We thank M. Müller for supplying the expression vector containing KtnC and D. Sherman for supplying the expression vector containing RhFRed. We thank E. A. Meucci, E. C. Bornowski and J. B. Pyser for assistance with the synthesis of substrates and C.-H. Chiang for help with acquisition of circular dichroism spectra.

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  1. These authors contributed equally: Lara E. Zetzsche, Jessica A. Yazarians

Authors and Affiliations

  1. Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA

    Lara E. Zetzsche, Jessica A. Yazarians, Suman Chakrabarty, Meagan E. Hinze, Lauren A. M. Murray, April L. Lukowski & Alison R. H. Narayan

  2. Program in Chemical Biology, University of Michigan, Ann Arbor, MI, USA

    Lara E. Zetzsche, April L. Lukowski & Alison R. H. Narayan

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

    Jessica A. Yazarians & Alison R. H. Narayan

  4. Arrowhead Pharmaceuticals, Inc., Madison, WI, USA

    Leo A. Joyce

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Contributions

L.E.Z., J.A.Y., S.C., M.E.H. and A.R.H.N. designed experiments and wrote the manuscript with feedback from all authors. L.E.Z. and A.L.L. generated yeast strains used in this work. L.E.Z. performed protein engineering and generated SSNs presented in this work. L.E.Z., J.A.Y., M.E.H. and L.A.M.M. conducted biocatalytic reactions. J.A.Y., S.C. and M.E.H. synthesized substrates and racemic product standards. S.C. carried out product isolation from preparative-scale biocatalytic reactions. L.A.J. calculated circular dichroism spectra and assigned the absolute configuration of products.

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Correspondence to Alison R. H. Narayan.

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Supplementary Information

This file contains the following sections: I. Chemical synthesis; II. Protein sequences, expression, and purification; III. Biocatalytic reactions with fungal P450 KtnC; IV. Directed evolution of KtnC; V. Biocatalytic reactions with P450–RhFRed enzymes; VI. Preparative-scale biocatalytic reactions; VII. Assignment of absolute configurations; VIII. 1H NMR and 13C NMR spectra of compounds; and IX. References.

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Zetzsche, L.E., Yazarians, J.A., Chakrabarty, S. et al. Biocatalytic oxidative cross-coupling reactions for biaryl bond formation. Nature 603, 79–85 (2022). https://doi.org/10.1038/s41586-021-04365-7

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