Cargando…

Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation †

Dinucleotide analogs of the messenger RNA cap (m(7)GpppN) are useful research tools and have potential applications as translational inhibitors or reagents for modification of in vitro transcribed mRNAs. It has been previously reported that replacing the methyl group at the N7-position with benzyl (...

Descripción completa

Detalles Bibliográficos
Autores principales: Wojcik, Radoslaw, Baranowski, Marek R., Markiewicz, Lukasz, Kubacka, Dorota, Bednarczyk, Marcelina, Baran, Natalia, Wojtczak, Anna, Sikorski, Pawel J., Zuberek, Joanna, Kowalska, Joanna, Jemielity, Jacek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8623273/
https://www.ncbi.nlm.nih.gov/pubmed/34834356
http://dx.doi.org/10.3390/pharmaceutics13111941
_version_ 1784605893236621312
author Wojcik, Radoslaw
Baranowski, Marek R.
Markiewicz, Lukasz
Kubacka, Dorota
Bednarczyk, Marcelina
Baran, Natalia
Wojtczak, Anna
Sikorski, Pawel J.
Zuberek, Joanna
Kowalska, Joanna
Jemielity, Jacek
author_facet Wojcik, Radoslaw
Baranowski, Marek R.
Markiewicz, Lukasz
Kubacka, Dorota
Bednarczyk, Marcelina
Baran, Natalia
Wojtczak, Anna
Sikorski, Pawel J.
Zuberek, Joanna
Kowalska, Joanna
Jemielity, Jacek
author_sort Wojcik, Radoslaw
collection PubMed
description Dinucleotide analogs of the messenger RNA cap (m(7)GpppN) are useful research tools and have potential applications as translational inhibitors or reagents for modification of in vitro transcribed mRNAs. It has been previously reported that replacing the methyl group at the N7-position with benzyl (Bn) produces a dinucleotide cap with superior properties. Here, we followed up on this finding by synthesizing 17 novel Bn(7)GpppG analogs and determining their structure–activity relationship regarding translation and translational inhibition. The compounds were prepared in two steps, including selective N7-alkylation of guanosine 5′-monophosphate by arylmethyl bromide followed by coupling with imidazole-activated GDP, with total yields varying from 22% to 62%. The compounds were then evaluated by determining their affinity for eukaryotic translation initiation factor 4E (eIF4E), testing their susceptibility to decapping pyrophosphatase, DcpS—which is most likely the major cellular enzyme targeting this type of compound—and determining their translation inhibitory properties in vitro. We also synthesized mRNAs capped with the evaluated compounds and tested their translational properties in A549 cells. Our studies identified N7-(4-halogenbenzyl) substituents as promising modifications in the contexts of either mRNA translation or translational inhibition. Finally, to gain more insight into the consequences at the molecular level of N7-benzylation of the mRNA cap, we determined the crystal structures of three compounds with eIF4E.
format Online
Article
Text
id pubmed-8623273
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-86232732021-11-27 Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation † Wojcik, Radoslaw Baranowski, Marek R. Markiewicz, Lukasz Kubacka, Dorota Bednarczyk, Marcelina Baran, Natalia Wojtczak, Anna Sikorski, Pawel J. Zuberek, Joanna Kowalska, Joanna Jemielity, Jacek Pharmaceutics Article Dinucleotide analogs of the messenger RNA cap (m(7)GpppN) are useful research tools and have potential applications as translational inhibitors or reagents for modification of in vitro transcribed mRNAs. It has been previously reported that replacing the methyl group at the N7-position with benzyl (Bn) produces a dinucleotide cap with superior properties. Here, we followed up on this finding by synthesizing 17 novel Bn(7)GpppG analogs and determining their structure–activity relationship regarding translation and translational inhibition. The compounds were prepared in two steps, including selective N7-alkylation of guanosine 5′-monophosphate by arylmethyl bromide followed by coupling with imidazole-activated GDP, with total yields varying from 22% to 62%. The compounds were then evaluated by determining their affinity for eukaryotic translation initiation factor 4E (eIF4E), testing their susceptibility to decapping pyrophosphatase, DcpS—which is most likely the major cellular enzyme targeting this type of compound—and determining their translation inhibitory properties in vitro. We also synthesized mRNAs capped with the evaluated compounds and tested their translational properties in A549 cells. Our studies identified N7-(4-halogenbenzyl) substituents as promising modifications in the contexts of either mRNA translation or translational inhibition. Finally, to gain more insight into the consequences at the molecular level of N7-benzylation of the mRNA cap, we determined the crystal structures of three compounds with eIF4E. MDPI 2021-11-16 /pmc/articles/PMC8623273/ /pubmed/34834356 http://dx.doi.org/10.3390/pharmaceutics13111941 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Wojcik, Radoslaw
Baranowski, Marek R.
Markiewicz, Lukasz
Kubacka, Dorota
Bednarczyk, Marcelina
Baran, Natalia
Wojtczak, Anna
Sikorski, Pawel J.
Zuberek, Joanna
Kowalska, Joanna
Jemielity, Jacek
Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation †
title Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation †
title_full Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation †
title_fullStr Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation †
title_full_unstemmed Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation †
title_short Novel N7-Arylmethyl Substituted Dinucleotide mRNA 5′ cap Analogs: Synthesis and Evaluation as Modulators of Translation †
title_sort novel n7-arylmethyl substituted dinucleotide mrna 5′ cap analogs: synthesis and evaluation as modulators of translation †
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8623273/
https://www.ncbi.nlm.nih.gov/pubmed/34834356
http://dx.doi.org/10.3390/pharmaceutics13111941
work_keys_str_mv AT wojcikradoslaw noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT baranowskimarekr noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT markiewiczlukasz noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT kubackadorota noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT bednarczykmarcelina noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT barannatalia noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT wojtczakanna noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT sikorskipawelj noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT zuberekjoanna noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT kowalskajoanna noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation
AT jemielityjacek noveln7arylmethylsubstituteddinucleotidemrna5capanalogssynthesisandevaluationasmodulatorsoftranslation