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Advanced siRNA Designs Further Improve In Vivo Performance of GalNAc-siRNA Conjugates
Open accessAbstract
Significant progress has been made in the advancement of RNAi therapeutics by combining a synthetic triantennary N-acetylgalactosamine ligand targeting the asialoglycoprotein receptor with chemically modified small interfering RNA (siRNA) designs, including the recently described Enhanced Stabilization Chemistry. This strategy has demonstrated robust RNAi-mediated gene silencing in liver after subcutaneous administration across species, including human. Here we demonstrate that substantial efficacy improvements can be achieved through further refinement of siRNA chemistry, optimizing the positioning of 2'-deoxy-2'-fluoro and 2'-O-methyl ribosugar modifications across both strands of the double-stranded siRNA duplex to enhance stability without compromising intrinsic RNAi activity. To achieve this, we employed an iterative screening approach across multiple siRNAs to arrive at advanced designs with low 2'-deoxy-2'-fluoro content that yield significantly improved potency and duration in preclinical species, including non-human primate. Liver exposure data indicate that the improvement in potency is predominantly due to increased metabolic stability of the siRNA conjugates.
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“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 [9]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
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“Next, we evaluated other siRNA design elements and con rmed that full chemical modi cation with strategic placement of 2'-uoro and 2'-O-methyl modi cations 9 is required for robust siRNA activity in the CNS.”
From Expanding RNAi therapeutics to extrahepatic tissues with lipophilic conjugates (2022) · cites this paper as [9]
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“2'-O-C16 was well tolerated across both strands, with potency similar to the unconjugated siRNA, except for a few positions known to be sensitive to sterically demanding 2'-modi cations 9 .”
From Expanding RNAi therapeutics to extrahepatic tissues with lipophilic conjugates (2022) · cites this paper as [9]
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Metadata
- Authors
- Foster, D. J., Brown, C. R., Shaikh, S., Trapp, C., Schlegel, M. K., Qian, K., Sehgal, A., Rajeev, K. G., Jadhav, V., Manoharan, M., Kuchimanchi, S., Maier, M. A., & Milstein, S.
- Affiliations
- Alnylam Pharmaceuticals (United States) (US)
- Corresponding author
- Milstein, Stuart
- ORCID
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2018-03
- Volume
- 26
- Issue
- 3
- Pages
- 708-717
- ISSN
- 1525-0016 (ISSN-L)
- PMC
- PMC5910670
- OpenAlex
- W2781906140
- Cited by
- 325
- References
- 32
- Citations by year
- 2026: 41 · 2025: 46 · 2024: 49 · 2023: 34 · 2022: 34 · 2021: 37 · 2020: 43 · 2019: 24 · 2018: 17
- FWCI
- 10.76
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- 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
- siRNA design, RNAi therapeutics, asialoglycoprotein receptor, siRNA stability, non-human primates, subcutaneous administration
- MeSH terms
- Acetylgalactosamine, Animals, Gene Expression Regulation, Liver, Male, Mice, Transgenic, RNA, Messenger, Gene Silencing, Hepatocytes, RNA Interference, RNA, Small Interfering, Mice, Argonaute Proteins
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