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Synthesis of a cytotoxic amanitin for biorthogonal conjugation

Synthesis of a cytotoxic amanitin for biorthogonal conjugation
Synthesis of a cytotoxic amanitin for biorthogonal conjugation

Alpha-amanitin is an exceedingly toxic, naturally occurring, bicyclic octapeptide that inhibits RNA polymerase and results in cellular and organismal death. Here we report the straightforward synthesis of an amanitin analogue that exhibited near-native toxicity. A pendant alkyne was readily installed to enable copper-catalyzed alkyne-azide cycloaddition (CuAAC) to azido-rhodamine and two azide-bearing versions of the RGD peptide. The fluorescent toxin analogue entered cells and provoked morphological changes consistent with cell death. The latter two conjugates are as toxic as the parent alkyne precursor, which demonstrates that conjugation does not diminish toxicity. In addition, we showed that toxicity depends on a single diastereomer of the unnatural amino acid, dihydroxyisoleucine (DHIle), at position 3. The convenient synthesis of a heptapeptide precursor now provides access to bioactive amanitin analogues that may be readily conjugated to biomolecules of interest.

amanitin, bioconjugates, click chemistry, natural products, peptides
1439-4227
1420-1425
Zhao, Liang
8a8924b8-aa93-4e73-91ea-5e9ae1db8afc
May, Jonathan P.
b54a262b-9f32-4435-8866-3b9c495294f3
Blanc, Antoine
15a2db80-abd7-4732-b4d8-0badc019dce9
Dietrich, David J.
4840d792-6898-4a9a-8a66-6148e49ecd47
Loonchanta, Anastak
161211a4-d472-4059-9f25-7a2838004195
Matinkhoo, Kaveh
3e5a94da-427c-4431-82bd-c39dfa4b6dd2
Pryyma, Alla
b4f09589-a5bb-4e34-a358-07a63c88b7a1
Perrin, David M.
12feb705-d3df-4f49-acf5-4283867ca47c
Zhao, Liang
8a8924b8-aa93-4e73-91ea-5e9ae1db8afc
May, Jonathan P.
b54a262b-9f32-4435-8866-3b9c495294f3
Blanc, Antoine
15a2db80-abd7-4732-b4d8-0badc019dce9
Dietrich, David J.
4840d792-6898-4a9a-8a66-6148e49ecd47
Loonchanta, Anastak
161211a4-d472-4059-9f25-7a2838004195
Matinkhoo, Kaveh
3e5a94da-427c-4431-82bd-c39dfa4b6dd2
Pryyma, Alla
b4f09589-a5bb-4e34-a358-07a63c88b7a1
Perrin, David M.
12feb705-d3df-4f49-acf5-4283867ca47c

Zhao, Liang, May, Jonathan P., Blanc, Antoine, Dietrich, David J., Loonchanta, Anastak, Matinkhoo, Kaveh, Pryyma, Alla and Perrin, David M. (2015) Synthesis of a cytotoxic amanitin for biorthogonal conjugation. ChemBioChem, 16 (10), 1420-1425. (doi:10.1002/cbic.201500226).

Record type: Article

Abstract

Alpha-amanitin is an exceedingly toxic, naturally occurring, bicyclic octapeptide that inhibits RNA polymerase and results in cellular and organismal death. Here we report the straightforward synthesis of an amanitin analogue that exhibited near-native toxicity. A pendant alkyne was readily installed to enable copper-catalyzed alkyne-azide cycloaddition (CuAAC) to azido-rhodamine and two azide-bearing versions of the RGD peptide. The fluorescent toxin analogue entered cells and provoked morphological changes consistent with cell death. The latter two conjugates are as toxic as the parent alkyne precursor, which demonstrates that conjugation does not diminish toxicity. In addition, we showed that toxicity depends on a single diastereomer of the unnatural amino acid, dihydroxyisoleucine (DHIle), at position 3. The convenient synthesis of a heptapeptide precursor now provides access to bioactive amanitin analogues that may be readily conjugated to biomolecules of interest.

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More information

Published date: 1 July 2015
Additional Information: Publisher Copyright: © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.
Keywords: amanitin, bioconjugates, click chemistry, natural products, peptides

Identifiers

Local EPrints ID: 451879
URI: http://eprints.soton.ac.uk/id/eprint/451879
ISSN: 1439-4227
PURE UUID: 02a2d05a-1424-4fd3-9914-c6d2292f9d76
ORCID for Jonathan P. May: ORCID iD orcid.org/0000-0003-1651-130X

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Date deposited: 02 Nov 2021 17:42
Last modified: 09 Jan 2022 04:05

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Contributors

Author: Liang Zhao
Author: Jonathan P. May ORCID iD
Author: Antoine Blanc
Author: David J. Dietrich
Author: Anastak Loonchanta
Author: Kaveh Matinkhoo
Author: Alla Pryyma
Author: David M. Perrin

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