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Enantioselective oxidation of unactivated C–H bonds in cyclic amines by iterative docking-guided mutagenesis of P450BM3 (CYP102A1)

Nature Synthesis volume 1, pages 936–945 (2022)Cite this article

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Abstract

Selective oxidation of ring C–H bonds is an attractive route to functionalized cyclic amines, which are versatile intermediates in drug synthesis and important fragment molecules in drug discovery. Here we report a combined substrate and enzyme engineering approach to achieve enantioselective functionalization of all unactivated C–H bonds of azepane, azocane, 7-azabicyclo[2.2.1]heptane and 8-azaspiro[4.5]decane by cytochrome P450BM3 (CYP102A1). Different N-modifying groups provide product diversity at high enantioselectivity (up to 99% e.e.) from a panel of just 48 variants of P450BM3. Substrate docking into molecular-dynamics-simulated structures of enzyme variants is shown to be useful for designing mutations to increase enantioselectivity by disfavouring binding poses leading to the unwanted enantiomer, and to increase enzymatic activity by disfavouring non-productive poses from ten or so variants per generation. The synthetic application of remote C–H activation within cyclic amines is exemplified by the synthesis of anisodamine via enantioselective hydroxylation of N-Boc-nortropinone.

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Fig. 1: Synthetic applications of enantioselective oxidation of cyclic amines by engineered P450BM3.
Fig. 2: Molecular structure of two cyclic amine oxidation products determined by single-crystal X-ray diffraction.
Fig. 3: IDGM of P450BM3 for the selective (S)-β oxidation of N-Ips-azepane (2).
Fig. 4: IDGM for (S)-endo oxidation of N-Boc-7-azabicyclo[2.2.1]heptane (5).

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Data availability

All data supporting the findings of this study are available within the paper and the Supplementary Information. Source data are provided with this paper. Crystallographic data for the structures reported in this Article have been deposited at the Cambridge Crystallographic Data Centre, under deposition numbers CCDC 2159261 ((R)-4b) and 2159262 ((S)-6a). Copies of the data can be obtained free of charge via https://www.ccdc.cam.ac.uk/structures/.

References

  1. Vitaku, E., Smith, D. T. & Njardarson, J. T. Analysis of the structural diversity, substitution patterns, and frequency of nitrogen heterocycles among US FDA approved pharmaceuticals. J. Med. Chem. 57, 10257–10274 (2014).

    Article  CAS  Google Scholar 

  2. Duke, S. O. et al. Natural toxins for use in pest management. Toxins 2, 1943–1962 (2010).

    Article  CAS  Google Scholar 

  3. Murray, C. W. & Rees, D. C. The rise of fragment-based drug discovery. Nat. Chem. 1, 187–192 (2009).

    Article  CAS  Google Scholar 

  4. Taylor, R. D., MacCoss, M. & Lawson, A. D. Rings in drugs. J. Med. Chem. 57, 5845–5859 (2014).

    Article  CAS  Google Scholar 

  5. Wijdeven, M. A., Willemsen, J. & Rutjes, F. P. J. T. The 3-hydroxypiperidine skeleton: key element in natural product synthesis. Eur. J. Org. Chem. 2010, 2831–2844 (2010).

    Article  Google Scholar 

  6. Nadin, A., Hattotuwagama, C. & Churcher, I. Lead-oriented synthesis: a new opportunity for synthetic chemistry. Angew. Chem. Int. Ed. 51, 1114–1122 (2012).

    Article  CAS  Google Scholar 

  7. Vo, C. V. & Bode, J. W. Synthesis of saturated N-heterocycles. J. Org. Chem. 79, 2809–2815 (2014).

    Article  CAS  Google Scholar 

  8. Allwood, D. M., Blakemore, D. C., Brown, A. D. & Ley, S. V. Metal-free coupling of saturated heterocyclic sulfonylhydrazones with boronic acids. J. Org. Chem. 79, 328–338 (2014).

    Article  CAS  Google Scholar 

  9. Yang, Y., Lan, J. & You, J. Oxidative C–H/C–H coupling reactions between two (hetero)arenes. Chem. Rev. 117, 8787–8863 (2017).

    Article  CAS  Google Scholar 

  10. Mitchell, E. A., Peschiulli, A., Lefevre, N., Meerpoel, L. & Maes, B. U. Direct α-functionalization of saturated cyclic amines. Chem. Eur. J. 18, 10092–10142 (2012).

    Article  CAS  Google Scholar 

  11. Shaw, M. H., Shurtleff, V. W., Terrett, J. A., Cuthbertson, J. D. & MacMillan, D. W. Native functionality in triple catalytic cross-coupling: sp3 C–H bonds as latent nucleophiles. Science 352, 1304–1308 (2016).

    Article  CAS  Google Scholar 

  12. Chen, W., Ma, L., Paul, A. & Seidel, D. Direct α-C–H bond functionalization of unprotected cyclic amines. Nat. Chem. 10, 165–169 (2018).

    Article  CAS  Google Scholar 

  13. Lennox, A. J. J. et al. Electrochemical aminoxyl-mediated α-cyanation of secondary piperidines for pharmaceutical building block diversification. J. Am. Chem. Soc. 140, 11227–11231 (2018).

    Article  CAS  Google Scholar 

  14. Millet, A., Larini, P., Clot, E. & Baudoin, O. Ligand-controlled β-selective C(sp3)–H arylation of N-Boc-piperidines. Chem. Sci. 4, 2241–2247 (2013).

    Article  CAS  Google Scholar 

  15. Millet, A., Dailler, D., Larini, P. & Baudoin, O. Ligand-controlled α- and β-arylation of acyclic N-Boc amines. Angew. Chem. Int. Ed. 53, 2678–2682 (2014).

    Article  CAS  Google Scholar 

  16. Topczewski, J. J., Cabrera, P. J., Saper, N. I. & Sanford, M. S. Palladium-catalysed transannular C–H functionalization of alicyclic amines. Nature 531, 220–224 (2016).

    Article  CAS  Google Scholar 

  17. Zhang, J., Park, S. & Chang, S. Catalytic access to bridged sila-N-heterocycles from piperidines via cascade sp3 and sp2 C–Si bond formation. J. Am. Chem. Soc. 140, 13209–13213 (2018).

    Article  CAS  Google Scholar 

  18. Chen, W., Paul, A., Abboud, K. A. & Seidel, D. Rapid functionalization of multiple C–H bonds in unprotected alicyclic amines. Nat. Chem. 12, 545–550 (2020).

    Article  CAS  Google Scholar 

  19. Oeschger, R. et al. Diverse functionalization of strong alkyl C–H bonds by undirected borylation. Science 368, 736–741 (2020).

    Article  CAS  Google Scholar 

  20. Trindade, A. F., Faulkner, E. L., Leach, A. G., Nelson, A. & Marsden, S. P. Fragment-oriented synthesis: β-elaboration of cyclic amine fragments using enecarbamates as platform intermediates. Chem. Commun. 56, 8802–8805 (2020).

    Article  CAS  Google Scholar 

  21. Holland, H. L., Morris, T. A., Nava, P. J. & Zabic, M. A new paradigm for biohydroxylation by Beauveria bassiana ATCC 7159. Tetrahedron 55, 7441–7460 (1999).

    Article  CAS  Google Scholar 

  22. Grogan, G. J. & Holland, H. L. The biocatalytic reactions of Beauveria spp. J. Mol. Catal. B 9, 1–32 (2000).

    Article  CAS  Google Scholar 

  23. de Raadt, A., Griengl, H. & Weber, H. The concept of docking and protecting groups in biohydroxylation. Chem. Eur. J. 7, 27–31 (2001).

    Article  Google Scholar 

  24. Sawayama, A. M. et al. A panel of cytochrome P450 BM3 variants to produce drug metabolites and diversify lead compounds. Chem. Eur. J. 15, 11723–11729 (2009).

    Article  CAS  Google Scholar 

  25. Reinen, J. et al. Efficient screening of cytochrome P450 BM3 mutants for their metabolic activity and diversity toward a wide set of drug-like molecules in chemical space. Drug Metab. Dispos. 39, 1568–1576 (2011).

    Article  CAS  Google Scholar 

  26. Ren, X. et al. Drug oxidation by cytochrome P450BM3: metabolite synthesis and discovering new P450 reaction types. Chem. Eur. J. 21, 15039–15047 (2015).

    Article  CAS  Google Scholar 

  27. Fasan, R. Tuning P450 enzymes as oxidation catalysts. ACS Catal. 2, 647–666 (2012).

    Article  CAS  Google Scholar 

  28. Roiban, G. D. & Reetz, M. T. Expanding the toolbox of organic chemists: directed evolution of P450 monooxygenases as catalysts in regio- and stereoselective oxidative hydroxylation. Chem. Commun. 51, 2208–2224 (2015).

    Article  CAS  Google Scholar 

  29. Wei, Y., Ang, E. L. & Zhao, H. Recent developments in the application of P450 based biocatalysts. Curr. Opin. Chem. Biol. 43, 1–7 (2018).

    Article  CAS  Google Scholar 

  30. Urlacher, V. B. & Girhard, M. Cytochrome P450 monooxygenases in biotechnology and synthetic biology. Trends Biotechnol. 37, 882–897 (2019).

    Article  CAS  Google Scholar 

  31. Pham, S. Q., Pompidor, G., Liu, J., Li, X. D. & Li, Z. Evolving P450pyr hydroxylase for highly enantioselective hydroxylation at non-activated carbon atom. Chem. Commun. 48, 4618–4620 (2012).

    Article  CAS  Google Scholar 

  32. Miura, Y. & Fulco, A. J. ω-1, ω-2 and ω-3 hydroxylation of long-chain fatty acids, amides and alcohols by a soluble enzyme system from Bacillus megaterium. Biochim. Biophys. Acta 388, 305–317 (1975).

    Article  CAS  Google Scholar 

  33. Whitehouse, C. J. C., Bell, S. G. & Wong, L. L. P450BM3 (CYP102A1): connecting the dots. Chem. Soc. Rev. 41, 1218–1260 (2012).

    Article  CAS  Google Scholar 

  34. Glieder, A., Farinas, E. T. & Arnold, F. H. Laboratory evolution of a soluble, self-sufficient, highly active alkane hydroxylase. Nat. Biotechnol. 20, 1135–1139 (2002).

    Article  CAS  Google Scholar 

  35. Reetz, M. T., Bocola, M., Carballeira, J. D., Zha, D. & Vogel, A. Expanding the range of substrate acceptance of enzymes: combinatorial active-site saturation test. Angew. Chem. Int. Ed. 44, 4192–4196 (2005).

    Article  CAS  Google Scholar 

  36. Jung, S. T., Lauchli, R. & Arnold, F. H. Cytochrome P450: taming a wild type enzyme. Curr. Opin. Biotechnol. 22, 809–817 (2011).

    Article  CAS  Google Scholar 

  37. Kille, S., Zilly, F. E., Acevedo, J. P. & Reetz, M. T. Regio- and stereoselectivity of P450-catalysed hydroxylation of steroids controlled by laboratory evolution. Nat. Chem. 3, 738–743 (2011).

    Article  CAS  Google Scholar 

  38. Seifert, A., Antonovici, M., Hauer, B. & Pleiss, J. An efficient route to selective bio-oxidation catalysts: an iterative approach comprising modeling, diversification, and screening, based on CYP102A1. ChemBioChem 12, 1346–1351 (2011).

    Article  CAS  Google Scholar 

  39. Zhang, K., El Damaty, S. & Fasan, R. P450 fingerprinting method for rapid discovery of terpene hydroxylating P450 catalysts with diversified regioselectivity. J. Am. Chem. Soc. 133, 3242–3245 (2011).

    Article  CAS  Google Scholar 

  40. Zhang, K., Shafer, B. M., Demars, M. D. 2nd, Stern, H. A. & Fasan, R. Controlled oxidation of remote sp3 C–H bonds in artemisinin via P450 catalysts with fine-tuned regio- and stereoselectivity. J. Am. Chem. Soc. 134, 18695–18704 (2012).

    Article  CAS  Google Scholar 

  41. Narayan, A. R. et al. Enzymatic hydroxylation of an unactivated methylene C–H bond guided by molecular dynamics simulations. Nat. Chem. 7, 653–660 (2015).

    Article  CAS  Google Scholar 

  42. Loskot, S. A., Romney, D. K., Arnold, F. H. & Stoltz, B. M. Enantioselective total synthesis of nigelladine A via late-stage C–H oxidation enabled by an engineered P450 enzyme. J. Am. Chem. Soc. 139, 10196–10199 (2017).

    Article  CAS  Google Scholar 

  43. Acevedo-Rocha, C. G. et al. P450-catalyzed regio- and diastereoselective steroid hydroxylation: efficient directed evolution enabled by mutability landscaping. ACS Catal. 8, 3395–3410 (2018).

    Article  CAS  Google Scholar 

  44. Li, Y. & Wong, L. L. Multi-functional oxidase activity of CYP102A1 (P450BM3) in the oxidation of quinolines and tetrahydroquinolines. Angew. Chem. Int. Ed. 58, 9551–9555 (2019).

    Article  CAS  Google Scholar 

  45. Li, A. et al. Regio- and stereoselective steroid hydroxylation at the C7-position by cytochrome P450 monooxygenase mutants. Angew. Chem. Int. Ed. 59, 12499–12505 (2020).

    Article  CAS  Google Scholar 

  46. Chen, W., Fisher, M. J., Leung, A., Cao, Y. & Wong, L. L. Oxidative diversification of steroids by nature-inspired scanning glycine mutagenesis of P450BM3 (CYP102A1). ACS Catal. 10, 8334–8343 (2020).

    Article  CAS  Google Scholar 

  47. Paulsen, M. D. & Ornstein, R. L. Dramatic differences in the motions of the mouth of open and closed cytochrome P450BM3 by molecular dynamics simulations. Proteins Struct. Funct. Genet. 21, 237–243 (1995).

    Article  CAS  Google Scholar 

  48. Pleiss, J. Systematic analysis of large enzyme families: identification of specificity- and selectivity-determining hotspots. ChemCatChem 6, 944–950 (2014).

    Article  CAS  Google Scholar 

  49. Dodani, S. C. et al. Discovery of a regioselectivity switch in nitrating P450s guided by molecular dynamics simulations and Markov models. Nat. Chem. 8, 419–425 (2016).

    Article  CAS  Google Scholar 

  50. Capoferri, L. et al. Insights into regioselective metabolism of mefenamic acid by cytochrome P450 BM3 mutants through crystallography, docking, molecular dynamics, and free energy calculations. Proteins Struct. Funct. Bioinf. 84, 383–396 (2016).

    Article  CAS  Google Scholar 

  51. Ebert, M. C. C. J. C., Espinola, J. G., Lamoureux, G. & Pelletier, J. N. Substrate-specific screening for mutational hotspots using biased molecular dynamics simulations. ACS Catal. 7, 6786–6797 (2017).

    Article  CAS  Google Scholar 

  52. Petrovic, D., Bokel, A., Allan, M., Urlacher, V. B. & Strodel, B. Simulation-guided design of cytochrome P450 for chemo- and regioselective macrocyclic oxidation. J. Chem. Inf. Model. 58, 848–858 (2018).

    Article  CAS  Google Scholar 

  53. Li, Z. et al. Engineering cytochrome P450 enzyme systems for biomedical and biotechnological applications. J. Biol. Chem. 295, 833–849 (2020).

    Article  Google Scholar 

  54. Peng, Y. Q. et al. A chemoenzymatic strategy for the synthesis of steroid drugs enabled by P450 monooxygenase-mediated steroidal core modification. ACS Catal. 12, 2907–2914 (2022).

    Article  CAS  Google Scholar 

  55. Whitehouse, C. J. C. et al. Evolved CYP102A1 (P450BM3) variants oxidise a range of non-natural substrates and offer new selectivity options. Chem. Commun., 966–968 (2008).

  56. Whitehouse, C. J. C. et al. Structural basis for the properties of two single-site proline mutants of CYP102A1 (P450BM3). ChemBioChem 11, 2549–2556 (2010).

    Article  CAS  Google Scholar 

  57. Whitehouse, C. J. C. et al. Structure, electronic properties and catalytic behaviour of an activity-enhancing CYP102A1 (P450BM3) variant. Dalton Trans. 40, 10383–10396 (2011).

    Article  CAS  Google Scholar 

  58. Ren, X., O’Hanlon, J. A., Morris, M., Robertson, J. & Wong, L. L. Synthesis of imidazolidin-4-ones via a cytochrome P450-catalyzed intramolecular C–H amination. ACS Catal. 6, 6833–6837 (2016).

    Article  CAS  Google Scholar 

  59. O’Hanlon, J. A., Ren, X., Morris, M., Wong, L. L. & Robertson, J. Hydroxylation of anilides by engineered cytochrome P450BM3. Org. Biomol. Chem. 15, 8780–8787 (2017).

    Article  Google Scholar 

  60. Li, C., Liu, Y., Pei, X.-Q. & Wu, Z.-L. Stereo-complementary bioreduction of saturated N-heterocyclic ketones. Process Biochem. 56, 90–97 (2017).

    Article  CAS  Google Scholar 

  61. Sheehan, J. C. & Bloom, B. M. The synthesis of teloidinone and 6-hydroxytropinone. J. Am. Chem. Soc. 74, 3825–3828 (1952).

    Article  CAS  Google Scholar 

  62. Stoll, A., Becker, B. & Jucker, E. Synthesis of α-hydroxysuccinaldehyde and 3,6-dihydroxytropane. Helv. Chim. Acta 35, 1263–1269 (1952).

    Article  CAS  Google Scholar 

  63. Nedenskov, P. & Clauson-Kaas, N. Simplified preparation of 6-hydroxytropinone. Acta Chem. Scand. 8, 1295–1295 (1954).

    Article  CAS  Google Scholar 

  64. Yu, Y. T. et al. Effectiveness of anisodamine for the treatment of critically ill patients with septic shock: a multicentre randomized controlled trial. Crit. Care 25, 349 (2021).

    Article  Google Scholar 

  65. Majewski, M. & Lazny, R. Stereoselective synthesis of tropane alkaloids: physoperuvine and dihydroxytropanes. Synlett 785–786 (1996).

  66. Majewski, M., Lazny, R. & Ulaczyk, A. Enantioselective ring opening of tropinone. A new entry into tropane alkaloids. Can. J. Chem. 75, 754–761 (1997).

    Article  CAS  Google Scholar 

  67. Cramer, N., Laschat, S., Baro, A. & Frey, W. Enantioselective desymmetrization of tropinone derivatives by hydroboration. Synlett 2175–2177 (2003).

  68. Kulkarni, K., Zhao, A. Y., Purcell, A. W. & Perlmutter, P. The enantioselective total synthesis and unambiguous proof of the absolute stereochemistry of pervilleine C. Synlett 2209–2212 (2008).

  69. Mao, Z. Y., Huang, S. Y., Gao, L. H., Wang, A. E. & Huang, P. Q. A novel and versatile method for the enantioselective syntheses of tropane alkaloids. Sci. China Chem. 57, 252–264 (2014).

    Article  CAS  Google Scholar 

  70. Fodor, G. & Kovacs, O. The stereochemistry of the tropane alkaloids. 3. The configuration of scopolamine and of valeroidine. J. Chem. Soc. 2341–2344 (1953).

  71. Cramer, N., Laschat, S. & Baro, A. Enzymatic resolution of tropinone derivatives. Synlett 2178–2181 (2003).

  72. Niu, Y. Y. et al. The absolute configuration plays an important role in muscarinic activity of BGT-A and its analogs. Bioorgan. Med. Chem. 16, 10251–10256 (2008).

    Article  CAS  Google Scholar 

  73. Fodor, G. & Soti, F. Correlation of valeroidine with S-(−)-methoxysuccinic acid and of mono-tigloyltropane and ditigloyltropane-3.6-diol with its R-(+)-antimer. Tetrahedron Lett. 1917–1921 (1964).

  74. Munoz, M. A., Munoz, O. & Joseph-Nathan, P. Absolute configuration determination and conformational analysis of (−)-(3S,6S)-3α,6β-diacetoxytropane using vibrational circular dichroism and DFT techniques. Chirality 22, 234–241 (2010).

    CAS  Google Scholar 

  75. Fodor, G., Toth, J. & Vincze, I. W. Stereochemistry of tropane alkaloids. 13. The absolute configuration and a simplified syntheses of valeroidine. J. Chem. Soc. 3219–3221 (1961).

  76. Wu, T. et al. Preparative separation of four isomers of synthetic anisodamine by HPLC and diastereomer crystallization. Chirality 31, 11–20 (2019).

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the Biotechnology and Biological Sciences Research Council, UK (BB/V003445/1). Y.Z. acknowledges a University of Oxford–China Scholarship Council Graduate Scholarship. M.W. and P.H.-L. acknowledge the EPSRC Centre for Doctoral Training in Synthesis for Biology and Medicine (EP/L015838/1) for graduate studentships. For the purpose of open access, the authors have applied a creative commons attribution (CC BY) licence to any author-accepted manuscript version arising.

Author information

Authors and Affiliations

  1. Department of Chemistry, University of Oxford, Inorganic Chemistry Laboratory, Oxford, UK

    Yuan Zhang, Yushu Li, Ellie Jaques & Luet L. Wong

  2. Oxford Suzhou Centre for Advanced Research, Suzhou, P.R. China

    Ziyue Xiong, Jeremy Robertson & Luet L. Wong

  3. Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Oxford, UK

    Mary Wilson, Kirsten E. Christensen, Pol Hernández-Lladó & Jeremy Robertson

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Contributions

Y.Z. performed enzyme and substrate engineering, activity screening, preparative-scale substrate oxidations, product characterization, MD simulations, substrate docking and docking-guided mutagenesis, and wrote the paper. Z.X. performed the stereoselective synthesis of anisodamine and wrote the paper. Y.L., E.J. and P.H.-L. carried out activity screening and product characterization experiments. M.W. performed activity screening, product characterization and steps in the synthesis of anisodamine. K.E.C. determined the crystal structures to assign the absolute configurations of two products and wrote the paper. J.R. conceived and guided the project, designed the synthesis of anisodamine and wrote the paper. L.L.W. conceived and guided the project, designed the steps in IDGM and wrote the paper.

Corresponding authors

Correspondence to Kirsten E. Christensen, Jeremy Robertson or Luet L. Wong.

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Peer review information

Nature Synthesis thanks Rita Bernhardt, Rudi Fasan, Nicholas Turner and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Primary handling editor Alison Stoddart, in collaboration with the Nature Synthesis team.

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Supplementary information

Supplementary Information

Supplementary Methods, Figs. 1–27, Tables 1–17, product characterization data and NMR spectra.

Supplementary Data 1

Crystallographic data for compound 4b, CCDC 2159261.

Supplementary Data 2

Structure factors for 4b, CCDC 2159261.

Supplementary Data 3

Crystallographic data for compound 6a, CCDC 2159262.

Supplementary Data 4

Structure factors for compound 6a, CCDC 2159262.

Source data

Source Data Fig. 2

Original Ortep figures for crystal structures.

Source Data Fig. 3

Source data for Fig. 3a,b.

Source Data Fig. 3

Original ray-traced figures from Pymol for Fig. 3c–f.

Source Data Fig. 4

Source data for Fig. 4a and Fig. 4b.

Source Data Fig. 4

Original ray-traced figures from Pymol for Fig. 4c,d.

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Zhang, Y., Xiong, Z., Li, Y. et al. Enantioselective oxidation of unactivated C–H bonds in cyclic amines by iterative docking-guided mutagenesis of P450BM3 (CYP102A1). Nat. Synth 1, 936–945 (2022). https://doi.org/10.1038/s44160-022-00166-6

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