Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative Linkers
Co-delivery systems of siRNA and chemotherapeutic drugs have been developed as an attractive strategy to optimize the efficacy of chemotherapy towards cancer cells with multidrug resistance. In these typical systems, siRNAs are usually associated to drugs within a carrier but without covalent intera...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-06-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/11/2714 |
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| author | Florian Gauthier Jean-Rémi Bertrand Jean-Jacques Vasseur Christelle Dupouy Françoise Debart |
| author_facet | Florian Gauthier Jean-Rémi Bertrand Jean-Jacques Vasseur Christelle Dupouy Françoise Debart |
| author_sort | Florian Gauthier |
| collection | DOAJ |
| description | Co-delivery systems of siRNA and chemotherapeutic drugs have been developed as an attractive strategy to optimize the efficacy of chemotherapy towards cancer cells with multidrug resistance. In these typical systems, siRNAs are usually associated to drugs within a carrier but without covalent interactions with the risk of a premature release and degradation of the drugs inside the cells. To address this issue, we propose a covalent approach to co-deliver a siRNA-drug conjugate with a redox-responsive self-immolative linker prone to intracellular glutathione-mediated disulfide cleavage. Herein, we report the use of two disulfide bonds connected by a pentane spacer or a <i>p</i>-xylene spacer as self-immolative linker between the primary amine of the anticancer drug doxorubicin (Dox) and the 2′-position of one or two ribonucleotides in RNA. Five Dox-RNA conjugates were successfully synthesized using two successive thiol-disulfide exchange reactions. The Dox-RNA conjugates were annealed with their complementary strands and the duplexes were shown to form an A-helix sufficiently stable under physiological conditions. The enzymatic stability of Dox-siRNAs in human serum was enhanced compared to the unmodified siRNA, especially when two Dox are attached to siRNA. The release of native Dox and RNA from the bioconjugate was demonstrated under reducing conditions suggesting efficient linker disintegration. These results demonstrate the feasibility of making siRNA-drug conjugates via disulfide-based self-immolative linkers for potential therapeutic applications. |
| first_indexed | 2024-03-10T19:14:14Z |
| format | Article |
| id | doaj.art-a3fee217948b4875be220b3f2d45c708 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T19:14:14Z |
| publishDate | 2020-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-a3fee217948b4875be220b3f2d45c7082023-11-20T03:32:28ZengMDPI AGMolecules1420-30492020-06-012511271410.3390/molecules25112714Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative LinkersFlorian Gauthier0Jean-Rémi Bertrand1Jean-Jacques Vasseur2Christelle Dupouy3Françoise Debart4IBMM, University of Montpellier, CNRS, ENSCM, 34095 Montpellier, FranceMETSY UMR 9018 CNRS, Université Paris-Sud, Gustave Roussy, University Paris-Saclay, 94800 Villejuif Cedex, FranceIBMM, University of Montpellier, CNRS, ENSCM, 34095 Montpellier, FranceIBMM, University of Montpellier, CNRS, ENSCM, 34095 Montpellier, FranceIBMM, University of Montpellier, CNRS, ENSCM, 34095 Montpellier, FranceCo-delivery systems of siRNA and chemotherapeutic drugs have been developed as an attractive strategy to optimize the efficacy of chemotherapy towards cancer cells with multidrug resistance. In these typical systems, siRNAs are usually associated to drugs within a carrier but without covalent interactions with the risk of a premature release and degradation of the drugs inside the cells. To address this issue, we propose a covalent approach to co-deliver a siRNA-drug conjugate with a redox-responsive self-immolative linker prone to intracellular glutathione-mediated disulfide cleavage. Herein, we report the use of two disulfide bonds connected by a pentane spacer or a <i>p</i>-xylene spacer as self-immolative linker between the primary amine of the anticancer drug doxorubicin (Dox) and the 2′-position of one or two ribonucleotides in RNA. Five Dox-RNA conjugates were successfully synthesized using two successive thiol-disulfide exchange reactions. The Dox-RNA conjugates were annealed with their complementary strands and the duplexes were shown to form an A-helix sufficiently stable under physiological conditions. The enzymatic stability of Dox-siRNAs in human serum was enhanced compared to the unmodified siRNA, especially when two Dox are attached to siRNA. The release of native Dox and RNA from the bioconjugate was demonstrated under reducing conditions suggesting efficient linker disintegration. These results demonstrate the feasibility of making siRNA-drug conjugates via disulfide-based self-immolative linkers for potential therapeutic applications.https://www.mdpi.com/1420-3049/25/11/2714doxorubicinsiRNAconjugationdisulfide bondself-immolative linkerthiol-disulfide exchange |
| spellingShingle | Florian Gauthier Jean-Rémi Bertrand Jean-Jacques Vasseur Christelle Dupouy Françoise Debart Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative Linkers Molecules doxorubicin siRNA conjugation disulfide bond self-immolative linker thiol-disulfide exchange |
| title | Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative Linkers |
| title_full | Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative Linkers |
| title_fullStr | Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative Linkers |
| title_full_unstemmed | Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative Linkers |
| title_short | Conjugation of Doxorubicin to siRNA Through Disulfide-based Self-immolative Linkers |
| title_sort | conjugation of doxorubicin to sirna through disulfide based self immolative linkers |
| topic | doxorubicin siRNA conjugation disulfide bond self-immolative linker thiol-disulfide exchange |
| url | https://www.mdpi.com/1420-3049/25/11/2714 |
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