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Cited by 278RNA Interference and Gene DeliveryDNA and Nucleic Acid ChemistryRNA and protein synthesis mechanisms

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

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Nair, Jayaprakash K., Attarwala, Husain, Sehgal, Alfica, Wang, Qianfan, Aluri, Krishna, Zhang, Xuemei, Gao, Minggeng, Liu, Ju +16 more·2017·Nucleic Acids Research
DOI 10.1093/nar/gkx818PMC PMC5737438
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Abstract

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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  • The naturally occurring RNAi pathway can be harnessed in extrahepatic organs by building on nearly two decades of liver siRNA design optimization, but novel delivery approaches are needed.

    “The naturally occurring RNAi pathway can also be harnessed in extrahepatic organs by building on nearly two decades of efforts on liver siRNA design optimization [9][10][11][12] ; however, novel delivery approaches are needed.”

    From Expanding RNAi therapeutics to extrahepatic tissues with lipophilic conjugates (2022) · cites this paper as [11]

    The Introduction describes advances in siRNA design and chemistry and explains how GalNAc conjugation enables delivery to the liver. It does not establish a nearly two-decade period of liver optimization or address whether those designs can be applied to extrahepatic organs. The statement that novel delivery approaches are needed follows the citation rather than being directly attributed to it. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • The most clinically-advanced siRNA conjugate is the multi-valent GalNAc, which drives delivery to and silencing in the liver.

    “Indeed, the most clinically-advanced siRNA conjugate is the multi-valent N-acetylgalactosamine (GalNAc)21, which drives delivery to and silencing in the liver.”

    From A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system (2019) · cites this paper as [21]

    The Introduction (page unavailable) describes triantennary GalNAc as enabling efficient siRNA delivery to the liver, and the Results and Discussion report liver gene silencing. The Conclusion mentions programs progressing into clinical development, but the paper does not establish that GalNAc is the most clinically advanced siRNA conjugate compared with others. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

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Authors
Nair, J. K., Attarwala, H., Sehgal, A., Wang, Q., Aluri, K., Zhang, X., Gao, M., Liu, J., Indrakanti, R., Schofield, S., Kretschmer, P., Brown, C. R., Gupta, S., Willoughby, J. L. S., Boshar, J. A., Jadhav, V., Charisse, K., Zimmermann, T., Fitzgerald, K., Manoharan, M., Rajeev, K. G., Akinc, A., Hutabarat, R., & Maier, M. A.
Affiliations
Alnylam Pharmaceuticals (United States) (US)
Corresponding author
Maier, Martin A.
ORCID
Venue
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Published
2017-09-15
Volume
45
Issue
19
Pages
10969-10977
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
DOI
10.1093/nar/gkx818
PMC
PMC5737438
OpenAlex
W2756425540
Cited by
278
References
1
Citations by year
2026: 32 · 2025: 37 · 2024: 45 · 2023: 28 · 2022: 25 · 2021: 32 · 2020: 37 · 2019: 22 · 2018: 20
FWCI
8.83
Type
journal article
Language
en
Open access
Licence
Locations
  • Nucleic Acids Research (publishedVersion, cc-by-nc)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
Funders
Alnylam Pharmaceuticals
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
Keywords
nuclease resistance, 5' exonuclease, siRNA modification, RNAi therapeutics, liver targeting, asialoglycoprotein receptor, gene silencing, oligonucleotide therapeutics, non-human primates
MeSH terms
Acetylgalactosamine, Animals, Humans, Kidney, Liver, Male, Metabolic Clearance Rate, Mice, Inbred C57BL, Drug Delivery Systems, Area Under Curve, RNA Interference, RNA, Small Interfering

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