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Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates

Covalent attachment of a synthetic triantennary N-acetylagalactosamine (GalNAc) ligand to chemically modified siRNA has enabled asialoglycoprotein (ASGPR)-mediated targeted delivery of therapeutically active siRNAs to hepatocytes in vivo. This approach has become transformative for the delivery of R...

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Autores principales: Nair, Jayaprakash K., Attarwala, Husain, Sehgal, Alfica, Wang, Qianfan, Aluri, Krishna, Zhang, Xuemei, Gao, Minggeng, Liu, Ju, Indrakanti, Ramesh, Schofield, Sally, Kretschmer, Philip, Brown, Christopher R., Gupta, Swati, Willoughby, Jennifer L.S., Boshar, Julie A., Jadhav, Vasant, Charisse, Klaus, Zimmermann, Tracy, Fitzgerald, Kevin, Manoharan, Muthiah, Rajeev, Kallanthottathil G., Akinc, Akin, Hutabarat, Renta, Maier, Martin A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5737438/
https://www.ncbi.nlm.nih.gov/pubmed/28981809
http://dx.doi.org/10.1093/nar/gkx818
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author Nair, Jayaprakash K.
Attarwala, Husain
Sehgal, Alfica
Wang, Qianfan
Aluri, Krishna
Zhang, Xuemei
Gao, Minggeng
Liu, Ju
Indrakanti, Ramesh
Schofield, Sally
Kretschmer, Philip
Brown, Christopher R.
Gupta, Swati
Willoughby, Jennifer L.S.
Boshar, Julie A.
Jadhav, Vasant
Charisse, Klaus
Zimmermann, Tracy
Fitzgerald, Kevin
Manoharan, Muthiah
Rajeev, Kallanthottathil G.
Akinc, Akin
Hutabarat, Renta
Maier, Martin A.
author_facet Nair, Jayaprakash K.
Attarwala, Husain
Sehgal, Alfica
Wang, Qianfan
Aluri, Krishna
Zhang, Xuemei
Gao, Minggeng
Liu, Ju
Indrakanti, Ramesh
Schofield, Sally
Kretschmer, Philip
Brown, Christopher R.
Gupta, Swati
Willoughby, Jennifer L.S.
Boshar, Julie A.
Jadhav, Vasant
Charisse, Klaus
Zimmermann, Tracy
Fitzgerald, Kevin
Manoharan, Muthiah
Rajeev, Kallanthottathil G.
Akinc, Akin
Hutabarat, Renta
Maier, Martin A.
author_sort Nair, Jayaprakash K.
collection PubMed
description Covalent attachment of a synthetic triantennary N-acetylagalactosamine (GalNAc) ligand to chemically modified siRNA has enabled asialoglycoprotein (ASGPR)-mediated targeted delivery of therapeutically active siRNAs to hepatocytes in vivo. This approach has become transformative for the delivery of RNAi therapeutics as well as other classes of investigational oligonucleotide therapeutics to the liver. For efficient functional delivery of intact drug into the desired subcellular compartment, however, it is critical that the nucleic acids are stabilized against nucleolytic degradation. Here, we compared two siRNAs of the same sequence but with different modification pattern resulting in different degrees of protection against nuclease activity. In vitro stability studies in different biological matrices show that 5′-exonuclease is the most prevalent nuclease activity in endo-lysosomal compartments and that additional stabilization in the 5′-regions of both siRNA strands significantly enhances the overall metabolic stability of GalNAc–siRNA conjugates. In good agreement with in vitro findings, the enhanced stability translated into substantially improved liver exposure, gene silencing efficacy and duration of effect in mice. Follow-up studies with a second set of conjugates targeting a different transcript confirmed the previous results, provided additional insights into kinetics of RISC loading and demonstrated excellent translation to non-human primates.
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spelling pubmed-57374382018-01-09 Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates Nair, Jayaprakash K. Attarwala, Husain Sehgal, Alfica Wang, Qianfan Aluri, Krishna Zhang, Xuemei Gao, Minggeng Liu, Ju Indrakanti, Ramesh Schofield, Sally Kretschmer, Philip Brown, Christopher R. Gupta, Swati Willoughby, Jennifer L.S. Boshar, Julie A. Jadhav, Vasant Charisse, Klaus Zimmermann, Tracy Fitzgerald, Kevin Manoharan, Muthiah Rajeev, Kallanthottathil G. Akinc, Akin Hutabarat, Renta Maier, Martin A. Nucleic Acids Res Chemical Biology and Nucleic Acid Chemistry Covalent attachment of a synthetic triantennary N-acetylagalactosamine (GalNAc) ligand to chemically modified siRNA has enabled asialoglycoprotein (ASGPR)-mediated targeted delivery of therapeutically active siRNAs to hepatocytes in vivo. This approach has become transformative for the delivery of RNAi therapeutics as well as other classes of investigational oligonucleotide therapeutics to the liver. For efficient functional delivery of intact drug into the desired subcellular compartment, however, it is critical that the nucleic acids are stabilized against nucleolytic degradation. Here, we compared two siRNAs of the same sequence but with different modification pattern resulting in different degrees of protection against nuclease activity. In vitro stability studies in different biological matrices show that 5′-exonuclease is the most prevalent nuclease activity in endo-lysosomal compartments and that additional stabilization in the 5′-regions of both siRNA strands significantly enhances the overall metabolic stability of GalNAc–siRNA conjugates. In good agreement with in vitro findings, the enhanced stability translated into substantially improved liver exposure, gene silencing efficacy and duration of effect in mice. Follow-up studies with a second set of conjugates targeting a different transcript confirmed the previous results, provided additional insights into kinetics of RISC loading and demonstrated excellent translation to non-human primates. Oxford University Press 2017-11-02 2017-09-15 /pmc/articles/PMC5737438/ /pubmed/28981809 http://dx.doi.org/10.1093/nar/gkx818 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Chemical Biology and Nucleic Acid Chemistry
Nair, Jayaprakash K.
Attarwala, Husain
Sehgal, Alfica
Wang, Qianfan
Aluri, Krishna
Zhang, Xuemei
Gao, Minggeng
Liu, Ju
Indrakanti, Ramesh
Schofield, Sally
Kretschmer, Philip
Brown, Christopher R.
Gupta, Swati
Willoughby, Jennifer L.S.
Boshar, Julie A.
Jadhav, Vasant
Charisse, Klaus
Zimmermann, Tracy
Fitzgerald, Kevin
Manoharan, Muthiah
Rajeev, Kallanthottathil G.
Akinc, Akin
Hutabarat, Renta
Maier, Martin A.
Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates
title Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates
title_full Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates
title_fullStr Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates
title_full_unstemmed Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates
title_short Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc–siRNA conjugates
title_sort impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of galnac–sirna conjugates
topic Chemical Biology and Nucleic Acid Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5737438/
https://www.ncbi.nlm.nih.gov/pubmed/28981809
http://dx.doi.org/10.1093/nar/gkx818
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