The Enzyme-Free Release of Nucleotides from Phosphoramidates Depends Strongly on the Amino Acid.


Journal

Angewandte Chemie (International ed. in English)
ISSN: 1521-3773
Titre abrégé: Angew Chem Int Ed Engl
Pays: Germany
ID NLM: 0370543

Informations de publication

Date de publication:
02 11 2020
Historique:
received: 19 06 2020
revised: 04 08 2020
pubmed: 7 8 2020
medline: 23 12 2020
entrez: 7 8 2020
Statut: ppublish

Résumé

Phosphoramidates composed of an amino acid and a nucleotide analogue are critical metabolites of prodrugs, such as remdesivir. Hydrolysis of the phosphoramidate liberates the nucleotide, which can then be phosphorylated to become the pharmacologically active triphosphate. Enzymatic hydrolysis has been demonstrated, but a spontaneous chemical process may also occur. We measured the rate of enzyme-free hydrolysis for 17 phosphoramidates of ribonucleotides with amino acids or related compounds at pH 7.5. Phosphoramidates of proline hydrolyzed fast, with a half-life time as short as 2.4 h for Pro-AMP in ethylimidazole-containing buffer at 37 °C; 45-fold faster than Ala-AMP and 120-fold faster than Phe-AMP. Crystal structures of Gly-AMP, Pro-AMP, βPro-AMP and Phe-AMP bound to RNase A as crystallization chaperone showed how well the carboxylate is poised to attack the phosphoramidate, helping to explain this reactivity. Our results are significant for the design of new antiviral prodrugs.

Identifiants

pubmed: 32757352
doi: 10.1002/anie.202008665
pmc: PMC7436718
doi:

Substances chimiques

Amides 0
Amino Acids 0
Antiviral Agents 0
Nucleotides 0
Phosphoric Acids 0
remdesivir 3QKI37EEHE
Adenosine Monophosphate 415SHH325A
phosphoramidic acid 9Q189608GB
Ribonuclease, Pancreatic EC 3.1.27.5
Alanine OF5P57N2ZX

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

20154-20160

Subventions

Organisme : Volkswagen Foundation
Pays : International
Organisme : Deutsche Forschungsgemeinschaft
ID : project ID 364653263 - TRR 235
Pays : International

Informations de copyright

© 2020 The Authors. Published by Wiley-VCH GmbH.

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Auteurs

Dejana Jovanovic (D)

Institut für Organische Chemie, Universität Stuttgart, 70569, Stuttgart, Germany.

Peter Tremmel (P)

Institut für Organische Chemie, Universität Stuttgart, 70569, Stuttgart, Germany.

Pradeep S Pallan (PS)

Department of Biochemistry, Vanderbilt University, School of Medicine, Nashville, TN, 37232, USA.

Martin Egli (M)

Department of Biochemistry, Vanderbilt University, School of Medicine, Nashville, TN, 37232, USA.

Clemens Richert (C)

Institut für Organische Chemie, Universität Stuttgart, 70569, Stuttgart, Germany.

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Classifications MeSH