siRNA Conjugates Carrying Sequentially Assembled Trivalent <i>N-</i>Acetylgalactosamine Linked Through Nucleosides Elicit Robust Gene Silencing <i>In Vivo</i> in Hepatocytes

Matsuda, Shigeo; Keiser, Kristofer; Nair, Jayaprakash K.; Charissé, Klaus; Manoharan, Rajar M; Kretschmer, Philip; Peng, Chang G.; Kel’in, Alexander V.; Pachamuthu, Kandasamy; Willoughby, Jennifer L. S.; Liebow, Abigail; Querbes, William; Yucius, Kristina; Nguyen, Tuyen; Milstein, Stuart; Maier, Martin A.; Rajeev, Kallanthottathil G.; Manoharan, Muthiah

doi:10.1021/cb501028c

Cited by 186 publications

(154 citation statements)

References 21 publications

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“…The Alnylam group has further optimized the chemistry of the glycoconjugates, providing simpler alternatives for synthesis (259,260). Multivalent glycoconjugates have also been used to target novel uncharged siRNA entities (47) with the attainment of similar functional effects in cells and in vivo .…”

Section: Approaches To Deliverymentioning

confidence: 99%

“…In this context, it is interesting to compare effects of the siRNA glycoconjugates described above with those of recent types of siRNA LNPs. While both approaches can effectively silence expression of hepatic genes, the LNP system can do so a doses of 0.1 mg/kg as compared to >5 mg/kg for the glycoconjugate (175,259). A final issue concerns stability.…”

Section: Approaches To Deliverymentioning

confidence: 99%

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The delivery of therapeutic oligonucleotides

2016

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The oligonucleotide therapeutics field has seen remarkable progress over the last few years with the approval of the first antisense drug and with promising developments in late stage clinical trials using siRNA or splice switching oligonucleotides. However, effective delivery of oligonucleotides to their intracellular sites of action remains a major issue. This review will describe the biological basis of oligonucleotide delivery including the nature of various tissue barriers and the mechanisms of cellular uptake and intracellular trafficking of oligonucleotides. It will then examine a variety of current approaches for enhancing the delivery of oligonucleotides. This includes molecular scale targeted ligand-oligonucleotide conjugates, lipid- and polymer-based nanoparticles, antibody conjugates and small molecules that improve oligonucleotide delivery. The merits and liabilities of these approaches will be discussed in the context of the underlying basic biology.

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Section: Approaches To Deliverymentioning

confidence: 99%

Section: Approaches To Deliverymentioning

confidence: 99%

The delivery of therapeutic oligonucleotides

2016

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“…siRNA conjugates have emerged as an alternative to nanocarrier-mediated delivery (9)(10)(11)(12)(13)(14), offering the possibility of improving siRNA pharmacokinetics without requiring a more complex delivery vehicle. Alnylam Pharmaceuticals has demonstrated high gene silencing potency of a trivalent N-Acetylgalactosamine (GalNAc) siRNA conjugate, which binds with high specificity and affinity to the asialoglycoprotein receptor on hepatocytes (15,16).…”

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confidence: 99%

Lipophilic siRNA targets albumin in situ and promotes bioavailability, tumor penetration, and carrier-free gene silencing

Sarett

Werfel

Lee

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

108

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Clinical translation of therapies based on small interfering RNA (siRNA) is hampered by siRNA's comprehensively poor pharmacokinetic properties, which necessitate molecule modifications and complex delivery strategies. We sought an alternative approach to commonly used nanoparticle carriers by leveraging the long-lived endogenous serum protein albumin as an siRNA carrier. We synthesized siRNA conjugated to a diacyl lipid moiety (siRNA-L2), which rapidly binds albumin in situ. siRNA-L2, in comparison with unmodified siRNA, exhibited a 5.7-fold increase in circulation half-life, an 8.6-fold increase in bioavailability, and reduced renal accumulation. Benchmarked against leading commercial siRNA nanocarrier in vivo jetPEI, siRNA-L2 achieved 19-fold greater tumor accumulation and 46-fold increase in per-tumor-cell uptake in a mouse orthotopic model of human triple-negative breast cancer. siRNA-L2 penetrated tumor tissue rapidly and homogeneously; 30 min after i.v. injection, siRNA-L2 achieved uptake in 99% of tumor cells, compared with 60% for jetPEI. Remarkably, siRNA-L2 achieved a tumor:liver accumulation ratio >40:1 vs. <3:1 for jetPEI. The improved pharmacokinetic properties of siRNA-L2 facilitated significant tumor gene silencing for 7 d after two i.v. doses. Proof-of-concept was extended to a patient-derived xenograft model, in which jetPEI tumor accumulation was reduced fourfold relative to the same formulation in the orthotopic model. The siRNA-L2 tumor accumulation diminished only twofold, suggesting that the superior tumor distribution of the conjugate over nanoparticles will be accentuated in clinical situations. These data reveal the immense promise of in situ albumin targeting for development of translational, carrier-free RNAi-based cancer therapies.

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“…Additionally, recent studies with double stranded siRNA GalNAc conjugates have also demonstrated the need for appropriate positioning of trivalent GalNAc sugars for optimal activity. 32,33 In conclusion, single stranded chemically modified ASOs can be effectively delivered to hepatocytes using two and even a single GalNAc sugar. This can significantly reduce the structural complexity associated with manufacturing such conjugates for clinical applications.…”

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confidence: 94%