Discovery of long-acting APOC3 siRNA for treating patients with hypertriglyceridemia

Figures

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  Figure 1.. A. Structure of the modified nucleic acids and 5’-N-acetylgalactosamine (GalNAc) cluster conjugated sense strand used for developing long-acting APOC3 siRNAs; B. Numbers indicating nucleoside and backbone positions from the 5’-end of the antisense strand (As) and the sense strand (Ss) of siRNA.

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  Figure 2.. Duration of activity of hAPOC3 siRNA plozasiran surrogate 1 and 2 to reduce plasma apoC3 protein (A) and triglyceride (B) in huAPOC3 transgenic mice (n = 3) after a single dose of 1 mg kg−1 for up to 105 days. Dose-response comparison of siRNA 1 and 2 following a single dose of 1.0, 0.3, and 0.1 mg kg−1 with plasma apoC3 protein (C) and triglycerides (D) evaluated on day 14. Error bars represent standard deviation.

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  Figure 3.. (A). Duration of activity of hApoC3 siRNA 2 containing 5’-deoxy-(E)-5’-vinyl phosphonate (VP) and 3 containing 5’-deoxy-5’-methylene phosphonate (MP) to reduce plasma apoC3 protein in huAPOC3 transgenic mice (n = 3) after a single dose of 0.1, 0.3, 1 mg kg−1 for up to 98 days. (B). Dose response comparison of siRNA 2 and 3 of plasma apoC3 reductions at day 14. Error bars represent standard deviation.

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  Figure 4.. Duration of activity of hAPOC3 siRNA 2, 4-8 to reduce plasma apoC3 protein (A and C) and triglyceride (B and D) in huAPOC3 transgenic mice (n = 3) after a single dose of 1 mg kg−1 for up to 28 to 105 days. Error bars represent standard deviation.

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  Figure 5.. Duration of activity hAPOC3 siRNA 2, 9-14 to reduce plasma apoC3 protein (A and C) and triglyceride (B and D) in huAPOC3 transgenic mice (n = 3) after a single dose 1 mg kg−1 for up to 28–84 days. Error bars represent standard deviation.

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  Figure 6.. Duration of activity of hAPOC3 siRNA 2, 15-18 to reduce plasma apoC3 protein (A) and triglyceride (B) in huAPOC3 transgenic mice (n = 3) after a single dose of 1 mg kg−1 for up to 84 days. Dose-responsive duration of activity of hAPOC3 siRNA 17 (C) and 18 (D) to reduce plasma apoC3 protein in transgenic mice after a single dose of 1, 2, and 3 mg kg−1. Error bars represent standard deviation.

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  Figure 7.. Duration of activity of hAPOC3 siRNA 2, 19-26 to reduce plasma apoC3 protein (A) and triglyceride (B) in huAPOC3 transgenic mice (n = 3) after a single dose of 1 mg kg−1 for up to 84 days. Error bars represent standard deviation.

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  Figure 8.. (A). Duration of activity of siRNA 1 compared with lead hAPOC3 siRNAs 6, 17-18, and 26 after a single dose of 1 mg kg−1, with plasma apoC3 protein evaluated in huAPOC3 transgenic mice for up to 12 weeks. Composite data from multiple studies representing n = 3–6 for each siRNA. B–D. Dose-responsive activity of hAPOC3 siRNA 26 (0.01, 0.03, 0.1, and 1 mg kg−1 single dose) to reduce plasma liver mRNA (B), plasma ApoC3 protein (C), and plasma triglyceride (D) in transgenic mice. (E). ED50 values for liver apoC3 mRNA, plasma apoC3, and plasma triglyceride reductions following siRNA 26 treatment. Error bars represent standard deviation.

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  Figure 9.. (A). Volcano plot depicting global gene expression changes in primary human hepatocytes at 96 h after free uptake of 200, 1000, and 5000 nM of hAPOC3 siRNAs 2 and 26. Enrichment of the seed region complementary in 3’-UTRs is tabulated N: 2 technical replicates; (B). Plot of relative IC50 values of down-regulated genes relative to on-target IC50 values after free uptake of hAPOC3 siRNA 2 and 26.

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  Figure 10.. (A) Potency of hAPOC3 siRNA 26 in primary human hepatocytes. Error bars represent standard deviation; (B) Potency of hAPOC3 siRNA 26 in monkey hepatocytes. Error bars represent standard deviation; (C) Activity of hAPOC3 siRNA 26 in monkey liver dosed subcutaneously at 4 mg kg−1 and 50 mg kg−1 (n = 4). Error bars represent standard deviation.