Cargando…

Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach

Steric blocking antisense oligonucleotides (ASO) are promising tools for splice modulation such as exon-skipping, although their therapeutic effect may be compromised by insufficient delivery. To address this issue, we investigated the synthesis of a 20-mer 2′-OMe PS oligonucleotide conjugated at 3′...

Descripción completa

Detalles Bibliográficos
Autores principales: Marchesi, Elena, Bovolenta, Matteo, Preti, Lorenzo, Capobianco, Massimo L., Mamchaoui, Kamel, Bertoldo, Monica, Perrone, Daniela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8707236/
https://www.ncbi.nlm.nih.gov/pubmed/34946743
http://dx.doi.org/10.3390/molecules26247662
_version_ 1784622387604488192
author Marchesi, Elena
Bovolenta, Matteo
Preti, Lorenzo
Capobianco, Massimo L.
Mamchaoui, Kamel
Bertoldo, Monica
Perrone, Daniela
author_facet Marchesi, Elena
Bovolenta, Matteo
Preti, Lorenzo
Capobianco, Massimo L.
Mamchaoui, Kamel
Bertoldo, Monica
Perrone, Daniela
author_sort Marchesi, Elena
collection PubMed
description Steric blocking antisense oligonucleotides (ASO) are promising tools for splice modulation such as exon-skipping, although their therapeutic effect may be compromised by insufficient delivery. To address this issue, we investigated the synthesis of a 20-mer 2′-OMe PS oligonucleotide conjugated at 3′-end with ursodeoxycholic acid (UDCA) involved in the targeting of human DMD exon 51, by exploiting both a pre-synthetic and a solution phase approach. The two approaches have been compared. Both strategies successfully provided the desired ASO 51 3′-UDC in good yield and purity. It should be pointed out that the pre-synthetic approach insured better yields and proved to be more cost-effective. The exon skipping efficiency of the conjugated oligonucleotide was evaluated in myogenic cell lines and compared to that of unconjugated one: a better performance was determined for ASO 51 3′-UDC with an average 9.5-fold increase with respect to ASO 51.
format Online
Article
Text
id pubmed-8707236
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-87072362021-12-25 Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach Marchesi, Elena Bovolenta, Matteo Preti, Lorenzo Capobianco, Massimo L. Mamchaoui, Kamel Bertoldo, Monica Perrone, Daniela Molecules Article Steric blocking antisense oligonucleotides (ASO) are promising tools for splice modulation such as exon-skipping, although their therapeutic effect may be compromised by insufficient delivery. To address this issue, we investigated the synthesis of a 20-mer 2′-OMe PS oligonucleotide conjugated at 3′-end with ursodeoxycholic acid (UDCA) involved in the targeting of human DMD exon 51, by exploiting both a pre-synthetic and a solution phase approach. The two approaches have been compared. Both strategies successfully provided the desired ASO 51 3′-UDC in good yield and purity. It should be pointed out that the pre-synthetic approach insured better yields and proved to be more cost-effective. The exon skipping efficiency of the conjugated oligonucleotide was evaluated in myogenic cell lines and compared to that of unconjugated one: a better performance was determined for ASO 51 3′-UDC with an average 9.5-fold increase with respect to ASO 51. MDPI 2021-12-17 /pmc/articles/PMC8707236/ /pubmed/34946743 http://dx.doi.org/10.3390/molecules26247662 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
Marchesi, Elena
Bovolenta, Matteo
Preti, Lorenzo
Capobianco, Massimo L.
Mamchaoui, Kamel
Bertoldo, Monica
Perrone, Daniela
Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach
title Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach
title_full Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach
title_fullStr Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach
title_full_unstemmed Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach
title_short Synthesis and Exon-Skipping Properties of a 3′-Ursodeoxycholic Acid-Conjugated Oligonucleotide Targeting DMD Pre-mRNA: Pre-Synthetic versus Post-Synthetic Approach
title_sort synthesis and exon-skipping properties of a 3′-ursodeoxycholic acid-conjugated oligonucleotide targeting dmd pre-mrna: pre-synthetic versus post-synthetic approach
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8707236/
https://www.ncbi.nlm.nih.gov/pubmed/34946743
http://dx.doi.org/10.3390/molecules26247662
work_keys_str_mv AT marchesielena synthesisandexonskippingpropertiesofa3ursodeoxycholicacidconjugatedoligonucleotidetargetingdmdpremrnapresyntheticversuspostsyntheticapproach
AT bovolentamatteo synthesisandexonskippingpropertiesofa3ursodeoxycholicacidconjugatedoligonucleotidetargetingdmdpremrnapresyntheticversuspostsyntheticapproach
AT pretilorenzo synthesisandexonskippingpropertiesofa3ursodeoxycholicacidconjugatedoligonucleotidetargetingdmdpremrnapresyntheticversuspostsyntheticapproach
AT capobiancomassimol synthesisandexonskippingpropertiesofa3ursodeoxycholicacidconjugatedoligonucleotidetargetingdmdpremrnapresyntheticversuspostsyntheticapproach
AT mamchaouikamel synthesisandexonskippingpropertiesofa3ursodeoxycholicacidconjugatedoligonucleotidetargetingdmdpremrnapresyntheticversuspostsyntheticapproach
AT bertoldomonica synthesisandexonskippingpropertiesofa3ursodeoxycholicacidconjugatedoligonucleotidetargetingdmdpremrnapresyntheticversuspostsyntheticapproach
AT perronedaniela synthesisandexonskippingpropertiesofa3ursodeoxycholicacidconjugatedoligonucleotidetargetingdmdpremrnapresyntheticversuspostsyntheticapproach