Methods for treatment of APOC3-related diseases and disorders

Abstract

To provide methods for treatment of APOC3-related diseases and disorders.SOLUTION: Pharmaceutical compositions comprising APOC RNAi agents are administered to a human subject in need thereof according to methods disclosed herein to treat diseases and disorders associated with elevated triglyceride (TG) levels, such as familial chylomicronemia syndrome (FCS), hypertriglyceridemia, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, abnormal lipid and / or cholesterol metabolism, atherosclerosis, cardiovascular disease, coronary artery disease, hypertriglyceridemia induced pancreatitis, metabolic syndrome, type II diabetes mellitus, chylomicronemia, multifactorial chylomicronemia, or lipodystrophy syndromes including familial partial lipodystrophy.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 883,046, filed August 5, 2019, U.S. Provisional Patent Application No. 62 / 936,559, filed November 17, 2019, and U.S. Provisional Patent Application No. 62 / 970,613, filed February 5, 2020, the contents of each of which are hereby incorporated by reference in their entirety.

[0002] Sequence Listing This application contains a Sequence Listing which has been submitted in ASCII format and is hereby incorporated by reference in its entirety. This ASCII copy is entitled 30683_WO_SequenceListing.txt and is 3kb in size.

[0003] FIELD OF THEINVENTION Disclosed herein are methods for the treatment of diseases and disorders that may be mediated, at least in part, by silencing or inhibiting apolipoprotein C-III (also called APOC3, apoC-III, APOC-III, and APO C-III) gene expression using pharmaceutical compositions comprising an RNA interference (RNAi) agent that inhibits APOC3 gene expression. [Background technology]

[0004] background Apolipoprotein C-III (also called APOC3, apoC-III, APOC-III and APO C-III), encoded by the human apolipoprotein C-III gene, has recently emerged as a promising target for the treatment of diseases associated with hypertriglyceridemia. Elevated serum triglyceride (TG) levels have been identified as an independent risk factor for cardiovascular disease and as a contributing factor in the development of atherosclerosis. Individuals with severe hypertriglyceridemia (often >1000 mg / dL) are also at risk for recurrent pancreatitis. Triglycerides are primarily transported in the blood as major components of very low density lipoprotein cholesterol (VLDL-C) and chylomicron particles, known as TG-rich lipoproteins. Lipoproteins are composed of a hydrophobic triacylglycerol and cholesteryl ester core and a hydrophilic outer layer of phospholipids, cholesterol and apoproteins. APOC3 is one of these apoproteins.

[0005] APOC3 is mainly synthesized in the liver and plays an important role in the production, metabolism, and clearance of TG-rich lipoproteins from plasma. Several gain-of-function polymorphisms that are assumed to be contributing factors in the development of hypertriglyceridemia have been identified in the promoter region of the APOC3 gene (see, e.g., Wang, Y., et al., Association of Apolipoprotein C3 Genetic Polymorphisms with the Risk of Ischemic Stroke in the Northern Chinese Han Population, 11 PLoS One e0163910 (2016); Li, Y., et al., Apolipoprotein C3 gene variants and the risk of coronary heart disease: A meta-analysis 9 Meta Gene 104-109 (2016)). Increased APOC3 synthesis in the liver promotes the secretion of TG-rich VLDL-C. Moreover, the overabundance of APOC3 further increases serum TG levels by inhibiting the activity of lipoprotein lipase and hepatic lipase, slowing the catabolism of TG-rich lipoproteins. Moreover, elevated APOC3 also slows the hepatic clearance of TG-rich lipoproteins and their remnant particles by interfering with their binding to hepatic receptors.

[0006] Several large-scale genetic analysis studies have reported that individuals with loss-of-function mutations of APOC3 exhibit low levels of triglycerides and reduced incidence of cardiovascular disease. Furthermore, APOC3-deficient individuals also have increased HDL-C compared to individuals without APOC3 mutations (see, for example, Bernelot Moens, SJ, et al., Inhibition of ApoCIII: the next PCSK9? 25 Curr Opin Lipidol 418-422 (2014); Saleheen, D., et al., Human knockouts and phenotypic analysis in a cohort with a high rate of consanguinity, 544 Nature 235-239 (2017)). Currently, hypertriglyceridemia is often treated with one or more of niacin, fibrates, statins and fish oils in moderate cases, but in most cases, the reduction in serum TG is modest.In addition, available therapeutic drugs are often ineffective in patients with monogenic causes of very severe hypertriglyceridemia (e.g., patients with familial chylomicronemia syndrome (FCS)), because the majority of disease-causing mutations are in lipoprotein lipase (LPL), and mutations in cofactors or LPL-interacting proteins apolipoprotein C-II (APOC2), apolipoprotein AV (APOA5), lipase maturation factor 1 (LMF1), and glycosylphosphatidylinositol-anchored high density lipoprotein binding protein 1 (GPIHBP1) are observed less frequently. These mutations result in dysfunctional lipoprotein lipase, but functional lipoprotein lipase is required for optimal response to standard treatment.There is a need for effective therapeutic agents that can provide substantial TG-lowering effects for the treatment of diseases such as familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, lipodystrophy syndromes including familial partial lipodystrophy, obesity, dyslipidemia, nonalcoholic steatohepatitis, nonalcoholic fatty liver disease, hyperlipidemia, hypertriglyceridemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, and other dyslipidemia and metabolic-related disorders and diseases. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Wang, Y., et al., Association of Apolipoprotein C3 Genetic Polymorphisms with the Risk of Ischemic Stroke in the Northern Chinese Han Population, 11 PLoS One e0163910 (2016) [Non-Patent Document 2] Li, Y., et al., Apolipoprotein C3 gene variants and the risk of coronary heart disease: A meta-analysis 9 Meta Gene 104-109 (2016) [Non-Patent Document 3] Bernelot Moens, SJ, et al., Inhibition of ApoCIII: the next PCSK9? 25 Curr Opin Lipidol 418-422 (2014) [Non-Patent Document 4] Saleheen, D., et al., Human knockouts and phenotypic analysis in a cohort with a high rate of consanguinity, 544 Nature 235-239 (2017) Summary of the Invention [Means for solving the problem]

[0008] overview Described herein are methods of treating APOC3-related diseases and disorders in a human subject in need thereof, comprising administering to the human subject a pharmaceutical composition comprising a composition described in Table 2 (i.e., an APOC3 RNAi Drug Substance, also referred to herein as ADS-005), at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, wherein the pharmaceutical composition is administered subcutaneously and there is about one month between doses (i.e., monthly dosing). In some embodiments, the pharmaceutical composition used in the methods disclosed herein comprises, consists of, or consists essentially of a formulated APOC3 RNAi drug substance described in Table 3 (also referred to herein as ADS-005-1).

[0009] The APOC3 RNAi drug substance described herein provides unexpected efficacy and duration of effect.As shown in the data from human clinical studies presented herein, the APOC3 RNAi drug substance embodiment with a single dose of less than 100mg provides sustained inhibition of APOC3 gene expression, which lasts for several months and produces substantial reduction in triglycerides (TG) and other related lipid parameters, while maintaining a favorable safety profile.This has led to the novel discovery of low-dose and low-frequency dosing regimens for the treatment of APOC3-related diseases and disorders.

[0010] The methods of treatment described herein result in a reduction in APOC3 expression in a human subject, thereby resulting, inter alia, in a reduction in serum triglyceride (TG) levels in the subject.

[0011] Further described herein is a method of treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 (i.e., ADS-005) at a dose of between about 1 mg and about 50 mg, wherein the pharmaceutical composition is administered subcutaneously, and there is about one month between dose administrations (i.e., monthly dosing).

[0012] Further described herein is a method of treating an APOC3-associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein there are about 3 months between dose administrations (e.g., quarterly dosing or every 12 weeks dosing (q12w)).

[0013] Also described herein is a method of treating an APOC3-associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 50 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein there are about 3 months between dose administrations (e.g., quarterly dosing or every 12 weeks dosing (q12w)).

[0014] Described herein is a method of treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein a first dose is followed by a second dose about one month later, whereafter for subsequent doses there are about three months between dose administrations.

[0015] Described herein is a method of treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 50 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein a first dose is followed by a second dose about one month later, whereafter for subsequent doses there are about three months between dose administrations.

[0016] Further described herein is a method of treating an APOC3-associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein there are about 4 months between dose administrations.

[0017] Also described herein is a method of treating an APOC3-associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 50 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein there are about 4 months between dose administrations.

[0018] Described herein is a method of treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein a first dose is followed by a second dose about one month later, whereafter for subsequent doses there are about four months between dose administrations.

[0019] Described herein is a method of treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 50 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein a first dose is followed by a second dose about one month later, whereafter for subsequent doses there are about four months between dose administrations.

[0020] In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is between about 10 mg and about 100 mg. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is between about 10 mg and about 50 mg. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is between about 25 mg and about 50 mg. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is about 25 mg. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is about 50 mg. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is about 100 mg. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is about 10 mg. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is 100 mg or less. In some embodiments, the dose of the APOC3 RNAi drug substance administered in each dose is 50 mg or less. In some embodiments, the dose of APOC3 RNAi drug substance administered in each dose is 25 mg or less.

[0021] The treatment methods disclosed herein may provide substantial TG lowering effects for the treatment of APOC3-related diseases and disorders, such as hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, lipodystrophy syndromes including familial partial lipodystrophy, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, hypertriglyceridemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, and other dyslipidemia and metabolic-related disorders and diseases. The methods disclosed herein, in some embodiments, may treat APOC3-related diseases or disorders by substantially lowering TG levels, thereby reducing the risk of developing hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, lipodystrophy syndromes including familial partial lipodystrophy, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, hypertriglyceridemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, or other dyslipidemia or metabolic related disorders and diseases.

[0022] The pharmaceutical composition comprising APOC3 RNAi agent disclosed herein can be administered to human subject to inhibit the expression of APOC3 gene in the subject.In some embodiments, the subject is a human who has been previously diagnosed with elevated triglyceride level, overexpression of APOC3 protein, or one or more APOC3-related diseases or disorders.

[0023] Other objects, features, aspects and advantages of the present invention will become apparent from the following detailed description, the accompanying drawings and the claims. [Brief description of the drawings]

[0024] [Figure 1A]A chemical structure representation of the APOC3 RNAi drug substance described in Table 2 (referred to herein as ADS-005; i.e., an APOC3 RNAi agent conjugated to a tridentate N-acetyl-galactosamine targeting group at the 5' end of the sense strand) shown in free acid form. [Figure 1B] A chemical structure representation of the APOC3 RNAi drug substance described in Table 2 (referred to herein as ADS-005; i.e., an APOC3 RNAi agent conjugated to a tridentate N-acetyl-galactosamine targeting group at the 5' end of the sense strand) shown in free acid form. [Figure 1C] A chemical structure representation of the APOC3 RNAi drug substance described in Table 2 (referred to herein as ADS-005; i.e., an APOC3 RNAi agent conjugated to a tridentate N-acetyl-galactosamine targeting group at the 5' end of the sense strand) shown in free acid form. [Figure 1D] A chemical structure representation of the APOC3 RNAi drug substance described in Table 2 (referred to herein as ADS-005; i.e., an APOC3 RNAi agent conjugated to a tridentate N-acetyl-galactosamine targeting group at the 5' end of the sense strand) shown in free acid form.

[0025] [Figure 2A] Chemical structure representation of APOC3 RNAi drug substances listed in Table 2, shown in sodium salt form. [Figure 2B] Chemical structure representation of APOC3 RNAi drug substances listed in Table 2, shown in sodium salt form. [Figure 2C] Chemical structure representation of APOC3 RNAi drug substances listed in Table 2, shown in sodium salt form. [Figure 2D] Chemical structure representation of APOC3 RNAi drug substances listed in Table 2, shown in sodium salt form.

[0026] [Diagram 3]Schematic diagram of modified sense and antisense strands of the APOC3 RNAi drug substance described in Table 2 (referred to herein as ADS-005; i.e., an APOC3 RNAi agent conjugated to a tridentate N-acetyl-galactosamine targeting group at the 5' end of the sense strand). The following abbreviations are used in Figure 3: a, c, g, i, and u are 2'-O-methyl modified nucleotides (i represents inosine); Af, Cf, Gf, and Uf are 2'-fluoro (also referred to in the art as 2'-deoxy-2'-fluoro) modified nucleotides; o is a phosphodiester linkage; s is a phosphorothioate linkage; invAb is an inverted abasic residue or subunit; (NAG37)s has the following chemical structure: [ka] (shown in sodium salt form), [ka] (Shown in free acid form) is a tridentate N-acetyl-galactosamine targeting ligand having the formula:

[0027] [Figure 4] Phase I study design and dose escalation schedule (revised) for the Phase I clinical study described in Example 2.

[0028] [Diagram 5] Overview of the cohort for the Phase I clinical study (revised) described in Example 2.

[0029] [Figure 6]Graph showing serum APOC3 protein levels in healthy human volunteers administered placebo (all cohorts) or 25 mg of APOC3 RNAi drug substance (cohort 1) from the Phase I clinical study described in Example 2. As shown in Figures 6-16, the numbers in mg shown in the graphs for each respective figure refer to the dosed amount of APOC3 RNAi drug substance described in Table 2 (administered as the formulated APOC3 RNAi drug substance described in Table 3).

[0030] [Figure 7] Graph showing serum APOC3 protein levels in healthy human volunteers administered placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug substance (cohort 2) from the Phase I clinical study described in Example 2.

[0031] [Figure 8] Graph showing serum APOC3 protein levels in healthy human volunteers administered placebo (all cohorts) or a single 100 mg dose of APOC3 RNAi drug substance (cohort 3) from the Phase I clinical study described in Example 2.

[0032] [Figure 9] Graph showing serum APOC3 protein levels in healthy human volunteers administered placebo (all cohorts) or a single 10 mg dose of APOC3 RNAi drug substance (cohort 3) from the Phase I clinical study described in Example 2.

[0033] [Figure 10] Graph showing serum triglyceride (TG) levels in healthy human volunteers administered placebo (all cohorts) or a single 25 mg dose of APOC3 RNAi drug substance (cohort 1) from the Phase I clinical study described in Example 2.

[0034] [Figure 11]Graph showing serum triglyceride (TG) levels in healthy human volunteers administered placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug substance (cohort 2) from the Phase I clinical study described in Example 2.

[0035] [Figure 12] Graph showing serum triglyceride (TG) levels in healthy human volunteers administered placebo (all cohorts) or a single 100 mg dose of APOC3 RNAi drug substance (cohort 3) from the Phase I clinical study described in Example 2.

[0036] [Figure 13] Graph showing serum triglyceride (TG) levels in healthy human volunteers administered placebo (all cohorts) or a single 10 mg dose of APOC3 RNAi drug substance (cohort 4) from the Phase I clinical study described in Example 2.

[0037] [Figure 14] Graph showing serum very low density lipoprotein cholesterol (VLDL-C) levels in healthy human volunteers administered placebo (all cohorts) or a single 25 mg dose of APOC3 RNAi drug substance (cohort 1) from the Phase I clinical study described in Example 2.

[0038] [Figure 15] Graph showing serum VLDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug substance (cohort 2) from the Phase I clinical study described in Example 2.

[0039] [Figure 16] Graph showing serum VLDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 100 mg dose of APOC3 RNAi drug substance (cohort 3) from the Phase I clinical study described in Example 2.

[0040] [Figure 17] Graph showing serum VLDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 10 mg dose of APOC3 RNAi drug substance (cohort 4) from the Phase I clinical study described in Example 2.

[0041] [Figure 18] Graph showing serum very high density lipoprotein cholesterol (HDL-C) levels in healthy human volunteers administered placebo (all cohorts) or a single 25 mg dose of APOC3 RNAi drug substance (cohort 1) from the Phase I clinical study described in Example 2.

[0042] [Figure 19] Graph showing serum HDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug substance (cohort 2) from the Phase I clinical study described in Example 2.

[0043] [Figure 20] Graph showing serum HDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 100 mg dose of APOC3 RNAi drug substance (cohort 3) from the Phase I clinical study described in Example 2.

[0044] [Figure 21] Graph showing serum HDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 10 mg dose of APOC3 RNAi drug substance (cohort 4) from the Phase I clinical study described in Example 2.

[0045] [Figure 22]Graph showing serum low-density lipoprotein cholesterol (LDL-C) levels in healthy human volunteers administered placebo (all cohorts) or a single 25 mg dose of APOC3 RNAi drug substance (cohort 1) from the Phase I clinical study described in Example 2.

[0046] [Diagram 23] Graph showing serum LDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug substance (cohort 2) from the Phase I clinical study described in Example 2.

[0047] [Figure 24] Graph showing serum LDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 100 mg dose of APOC3 RNAi drug substance (cohort 3) from the Phase I clinical study described in Example 2.

[0048] [Diagram 25] Graph showing serum LDL-C levels in healthy human volunteers administered placebo (all cohorts) or a single 10 mg dose of APOC3 RNAi drug substance (cohort 4) from the Phase I clinical study described in Example 2.

[0049] [Figure 26] Graph showing serum APOC3 protein levels in chylomicronemia patients administered a 50 mg dose of APOC3 RNAi drug substance (cohort 5) from the Phase I clinical study described in Example 2.

[0050] [Figure 27] Graph showing serum triglyceride levels in chylomicronemic patients administered a 50 mg dose of APOC3 RNAi drug substance (Cohort 5) from the Phase I clinical study described in Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0051] Detailed Description RNAi agents The methods described herein include administering to a human subject a pharmaceutical composition, which includes a composition containing an RNA interference (RNAi) agent (referred to herein and in the art as an RNAi agent or RNAi trigger) capable of inhibiting the expression of the APOC3 gene. In some embodiments, the methods described herein include administering to a human subject a pharmaceutical composition, which includes an APOC3 RNAi drug substance (also referred to as ADS-005) described in Table 2. A suitable composition for use in the methods disclosed herein is composed of an RNAi agent that inhibits the expression of the APOC3 gene in a human subject and a targeting moiety or targeting group. In some embodiments, the RNAi agent includes a nucleotide sequence provided in Tables 1A and 1B, and the sense strand of the RNAi agent is further linked or conjugated to a targeting group that includes three N-acetyl-galactosamine targeting moieties (see, for example, Table B). An RNAi agent that inhibits the expression of the APOC3 gene in a human subject is referred to as an "APOC3 RNAi agent."

[0052] Generally, APOC3 RNAi agents include a sense strand (also called passenger strand) and an antisense strand (also called guide strand) that are annealed to form a duplex. APOC3 RNAi agents disclosed herein include RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecules that can degrade the messenger RNA (mRNA) transcript of APOC3 mRNA or inhibit its translation in a sequence-specific manner. APOC3 RNAi agents disclosed herein can operate via RNA interference mechanism (i.e., induce RNA interference via interaction with the RNA interference pathway machinery (RNA-induced silencing complex or RISC) of mammalian cells) or by any alternative mechanism(s) or pathway(s). Although APOC3 RNAi agents are believed to operate primarily via RNA interference mechanism as that term is used herein, the disclosed RNAi agents are not bound by or limited to any particular pathway or mechanism of action. RNAi agents are generally composed of a sense strand and an antisense strand, each 16-49 nucleotides in length, and include, but are not limited to, short or small interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), short hairpin RNA (shRNA), and Dicer substrates.

[0053] The length of the sense strand of an APOC3 RNAi agent is typically 16-49 nucleotides long, and the length of the antisense strand of an APOC3 RNAi agent is typically 18-49 nucleotides long. In some embodiments, the sense strand and the antisense strand are independently 17-26 nucleotides long. In some embodiments, the sense strand and the antisense strand are independently 21-26 nucleotides long. In some embodiments, the sense strand and the antisense strand are independently 21-24 nucleotides long. In some embodiments, the sense strand and / or the antisense strand are independently 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long. In some embodiments, the sense strand and the antisense strand are both 21 nucleotides long. The sense strand and the antisense strand can be either the same length or different lengths. The sense strand and the antisense strand can also form overhanging nucleotides at one or both ends of the APOC3 RNAi agent.

[0054] APOC3 RNAi agent inhibits, silences or knocks down APOC3 gene expression.As used herein, the terms "silencing", "reducing", "inhibiting", "downregulating" or "knockdown" refer to the expression of APOC3, and mean that the expression of the gene measured by the level of RNA transcribed from the gene or the level of polypeptide, protein or protein subunit translated from mRFNA in a cell, group of cells, tissue, organ or subject in which the gene is transcribed is reduced when the cell, group of cells, tissue, organ or subject is treated with RNAi agent, compared with a second cell, group of cells, tissue, organ or subject that is not treated in this way.In some examples, the reduction in gene expression is measured by comparing the baseline level of APOC3 mRNA or APOC3 protein in a human subject before administration of a composition comprising APOC3 RNAi agent with the APOC3 mRNA or APOC3 protein level after administration of a therapeutic agent.

[0055] The inhibition, silencing or knockdown of APOC3 gene can be measured by any suitable assay or method known in the art.The non-limiting examples shown herein, as well as the examples shown in International Patent Application Publication No. WO2019 / 051402 (Patent Application No. PCT / US2018 / 050248), the entirety of which is hereby incorporated by reference herein, provide certain examples of suitable assays for measuring APOC3 gene expression inhibition.The reference APOC3 mRNA gene transcript for normal humans (referred to as transcript variant 1; GenBank NM_000040.1) can be found in SEQ ID NO:1.

[0056] APOC3 RNAi agents suitable for use in the methods disclosed herein can be covalently linked or conjugated to a targeting group that includes one or more N-acetyl-galactosamine moieties. In some embodiments, APOC3 RNAi agents suitable for use in the methods disclosed herein are covalently linked or conjugated to a targeting group that includes one or more N-acetyl-galactosamine moieties, thereby forming an APOC3 RNAi drug substance as described in Table 2. In some embodiments, the methods described herein include administration of an APOC3 RNAi drug substance as described in Table 2. The APOC3 RNAi drug substances as described in Table 2 include the APOC3 RNAi agents shown in Table 1A (antisense strand) and Table 1B (sense strand). The N-acetyl-galactosamine moiety promotes targeting of the APOC3 RNAi agent to the asialoglycoprotein receptor (ASGPr) readily present on the surface of liver cells, which results in internalization of the APOC3 RNAi agent by endocytosis or other means.

[0057] APOC3 RNAi agents that may be suitable for use in the methods disclosed herein include antisense strands that have a region of complementarity to at least a portion of APOC3 mRNA. APOC3 RNAi agents and APOC3 RNAi drug substances suitable for use in the methods disclosed herein are described in the previously mentioned International Patent Application Publication No. WO2019 / 051402 (Patent Application No. PCT / US2018 / 050248), which is hereby incorporated by reference in its entirety.

[0058] As used herein, the terms "sequence" and "nucleotide sequence" refer to a sequence or order of nucleic acid bases or nucleotides described using a sequence of letters using standard terminology. As used herein, the terms "nucleobase" and "nucleotide" have the same meaning as commonly understood in the art.

[0059] As used herein, the term "complementary" when used to describe a first nucleotide sequence (e.g., the antisense strand of an RNAi agent) in relation to a second nucleotide sequence (e.g., the sense strand of an RNAi agent or a targeted mRNA sequence) refers to the ability of an oligonucleotide comprising the first nucleotide sequence to hybridize (form base pair hydrogen bonds) under mammalian physiological conditions (or other suitable conditions) and form a duplex or double helix structure with an oligonucleotide comprising the second nucleotide sequence under certain standard conditions. A person skilled in the art can select the most appropriate set of conditions for a hybridization test. Complementary sequences include Watson-Crick base pairs or non-Watson-Crick base pairs, and include natural or modified nucleotides or nucleotide mimetics, at least to the extent that the above hybridization requirements are met. Sequence identity or complementarity is independent of modification. For example, a and Af as defined herein are complementary to U (or T) and identical to A for purposes of determining identity or complementarity.

[0060] As used herein, "perfectly complementary" or "fully complementary" means that all (100%) of the bases in a contiguous sequence of a first oligonucleotide hybridize with the same number of nucleotides in a contiguous sequence of a second oligonucleotide. The contiguous sequence may comprise all or part of the first or second nucleotide sequence.

[0061] As used herein, "partially complementary" means that in a hybridized pair of nucleotide sequences, at least 70%, but not all, of the bases in a contiguous sequence of a first oligonucleotide hybridize to the same number of bases in a contiguous sequence of a second polynucleotide.

[0062] As used herein, "substantially complementary" means that in a hybridized pair of nucleotide sequences, at least 85% but not all of the bases in the contiguous sequence of the first oligonucleotide hybridize with the same number of bases in the contiguous sequence of the second polynucleotide.The terms "complementary", "fully complementary", "partially complementary" and "substantially complementary" are used herein with respect to the nucleotide correspondence between the sense strand and antisense strand of an RNAi agent, or between the antisense strand of an RNAi agent and the sequence of APOC3 mRNA.

[0063] As used herein, the term "substantially identical" or "substantially identical" when applied to a nucleic acid sequence means that a nucleic acid sequence comprises a sequence having at least about 85% or higher sequence identity compared to a reference sequence, for example, at least 90%, at least 95% or at least 99% identity. The percentage of sequence identity is determined by comparing two optimally aligned sequences over a comparison window. The percentage is calculated by determining the number of positions where identical nucleobases are present in both sequences to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the percentage of sequence identity. The invention disclosed herein encompasses nucleotide sequences that are substantially identical to those disclosed herein.

[0064] Modified Nucleotides and Modified Internucleoside Linkages APOC3 RNAi agents disclosed herein may be composed of modified nucleotides, which can maintain the activity of RNAi agents while simultaneously increasing serum stability, and can also minimize the possibility of activating interferon activity in humans.As used herein, "modified nucleotide" refers to nucleotides other than ribonucleotides (2'-hydroxyl nucleotides).In some embodiments, at least 50% of nucleotides (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100%) are modified nucleotides. As used herein, modified nucleotides include any known modified nucleotides known in the art, including, but not limited to, deoxyribonucleotides, nucleotide mimics, 2'-modified nucleotides, inverted nucleotides, modified nucleobase-containing nucleotides, bridged nucleotides, peptide nucleic acids (PNAs), 2',3'-seconucleotide mimics (unlocked nucleobase analogs), locked nucleotides, 3'-O-methoxy (2' internucleoside linked) nucleotides, 2'-F-arabinonucleotides, 5'-Me, 2'-fluoro nucleotides, morpholino nucleotides, vinylphosphonate-containing nucleotides, and cyclopropylphosphonate-containing nucleotides. In some embodiments, the modified nucleotides of the APOC3 RNAi agent are 2'-modified nucleotides (i.e., nucleotides having a group other than a hydroxyl group at the 2' position of the five-membered sugar ring). 2'-modified nucleotides include, but are not limited to, 2'-O-methyl nucleotides, 2'-deoxy-2'-fluoro nucleotides (commonly referred to simply as 2'-fluoro nucleotides), 2'-deoxy nucleotides, 2'-methoxyethyl (2'-O-2-methoxyethyl) nucleotides, 2'-amino nucleotides and 2'-alkyl nucleotides.Additional 2'-modified nucleotides are known in the art.Not all nucleotides in a given RNAi agent need to be uniformly modified.In addition, more than one modification can be incorporated into a single APOC3 RNAi agent, or even into that single nucleotide.The sense and antisense strands of an APOC3 RNAi agent can be synthesized and / or modified by methods known in the art such that a modification at one nucleotide is independent of a modification at another nucleotide.

[0065] In some embodiments, nucleobases (often simply referred to as "bases") can be modified. As commonly used in the art, natural nucleobases include the major purine bases adenine and guanine, and the major pyrimidine bases cytosine, thymine and uracil. Nucleobases can be modified to include, without limitation, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases and fluorinated bases (see, for example, Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008). The synthesis of such modified nucleobases (including phosphoramidite compounds that include modified nucleobases) is known in the art.

[0066] Modified nucleobases include, for example, 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, (e.g., 2-aminopropyladenine, 5-propynyluracil or 5-propynylcytosine), 5-methylcytosine (5-me-C), 5-hydroxymethylcytosine, inosine, xanthine, hypoxanthine, 2-aminoadenine, 6-alkyl (e.g., 6-methyl, 6-ethyl, 6-isopropyl or 6-n-butyl) derivatives of adenine and guanine, 2-alkyl (e.g., 2-methyl, 2-ethyl, 2-isopropyl or 2-n-butyl) and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine, 2- Included are thiocytosine, 5-halouracil, cytosine, 5-propynyluracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-sulfhydryl, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo (e.g., 5-bromo), 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine and 3-deazaadenine.

[0067] In some embodiments, all or substantially all of the nucleotides of APOC3 RNAi agent are modified nucleotides.As used herein, an RNAi agent in which substantially all of the nucleotides present are modified nucleotides is an RNAi agent that has 4 or fewer (i.e., 0, 1, 2, 3 or 4) nucleotides that are ribonucleotides (i.e., unmodified) in both sense strand and antisense strand.As used herein, a sense strand in which substantially all of the nucleotides present are modified nucleotides is a sense strand that has 2 or fewer (i.e., 0, 1 or 2) nucleotides that are ribonucleotides in the sense strand.As used herein, an antisense sense strand in which substantially all of the nucleotides present are modified nucleotides is an antisense strand that has 2 or fewer (i.e., 0, 1 or 2) nucleotides that are ribonucleotides in the sense strand.

[0068] In some embodiments, one or more nucleotides of an APOC3 RNAi agent are linked by a non-standard linkage or backbone (ie, a modified internucleoside linkage or modified backbone). Modified internucleoside linkages or backbones include, but are not limited to, phosphorothioate groups, chiral phosphorothioates, thiophosphates, phosphorodithioates, phosphotriesters, aminoalkyl-phosphotriesters, alkyl phosphonates (e.g., methyl phosphonates or 3'-alkylene phosphonates), chiral phosphonates, phosphinates, phosphoramidates (e.g., 3'-amino phosphoramidates, aminoalkyl phosphoramidates or thionophosphoramidates), thionoalkyl-phosphonates, thionoalkyl phosphotriesters, morpholino linkages, boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of boranophosphates, or boranophosphates having inverted polarity in which adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. In some embodiments, modified internucleoside linkages or backbones lack phosphorus atoms. Modified internucleoside linkages lacking phosphorus atoms include, but are not limited to, short chain alkyl or cycloalkyl intersugar linkages, mixed heteroatom and alkyl or cycloalkyl intersugar linkages, or one or more short chain heteroatom or heterocyclic intersugar linkages. In some embodiments, modified internucleoside backbones include, but are not limited to, siloxane backbones, sulfide backbones, sulfoxide backbones, sulfone backbones, formacetyl and thioformacetyl backbones, methyleneformacetyl and thioformacetyl backbones, alkene-containing backbones, sulfamate backbones, methyleneimino and methylenehydrazino backbones, sulfonate and sulfonamide backbones, amide backbones, and other backbones with mixed N, O, S, and CH2 components.

[0069] In some embodiments, the sense strand of an APOC3 RNAi agent may contain 1, 2, 3, 4, 5 or 6 phosphorothioate linkages, the antisense strand of an APOC3 RNAi agent may contain 1, 2, 3, 4, 5 or 6 phosphorothioate linkages, or both the sense strand and the antisense strand may independently contain 1, 2, 3, 4, 5 or 6 phosphorothioate linkages.In some embodiments, the sense strand of an APOC3 RNAi agent may contain 1, 2, 3 or 4 phosphorothioate linkages, the antisense strand of an APOC3 RNAi agent may contain 1, 2, 3 or 4 phosphorothioate linkages, or both the sense strand and the antisense strand may independently contain 1, 2, 3 or 4 phosphorothioate linkages.

[0070] In some embodiments, the sense strand of the APOC3 RNAi agent contains at least two phosphorothioate internucleoside linkages. In some embodiments, the at least two phosphorothioate internucleoside linkages are between nucleotides 1-3 from the 3' end of the sense strand. In some embodiments, the at least two phosphorothioate internucleoside linkages are between nucleotides 1-3, 2-4, 3-5, 4-6, 4-5, or 6-8 from the 5' end of the sense strand. In some embodiments, phosphorothioate internucleoside linkages are used to link the terminal nucleotide in the sense strand to a capping residue present at the 5' end, the 3' end, or both the 5' and 3' ends of the nucleotide sequence. In some embodiments, phosphorothioate internucleoside linkages are used to link a targeting group to the sense strand.

[0071] In some embodiments, the antisense strand of the APOC3 RNAi agent contains three or four phosphorothioate internucleoside linkages. In some embodiments, the antisense strand of the APOC3 RNAi agent contains three phosphorothioate internucleoside linkages. In some embodiments, the three phosphorothioate internucleoside linkages are between nucleotides 1-3 from the 5' end of the antisense strand and between nucleotides 19-21, 20-22, 21-23, 22-24, 23-25, or 24-26 from the 5' end of the antisense strand. In some embodiments, the APOC3 RNAi agent contains at least two phosphorothioate internucleoside linkages in the sense strand and three or four phosphorothioate internucleoside linkages in the antisense strand.

[0072] In some embodiments, an APOC3 RNAi agent contains one or more modified nucleotides and one or more modified internucleoside linkages. In some embodiments, a 2'-modified nucleoside is combined with a modified internucleoside linkage.

[0073] Capping residues or moieties In some embodiments, the sense strand may include one or more capping residues or moieties, which may be referred to in the art as "caps," "terminal caps," or "capping residues." As used herein, a "capping residue" is a non-nucleotide compound or other moiety that may be incorporated at one or more ends of the nucleotide sequence of an RNAi agent disclosed herein. The capping residue may, in some instances, provide the RNAi agent with certain beneficial properties, such as protection against exonuclease degradation. In some embodiments, an inverted abasic residue (invAb) (also referred to in the art as an "inverted abasic site") is added as a capping residue (see Table A) (see, e.g., F. Czauderna, Nucleic Acids Res., 2003, 31(11), 2705-16). Capping residues are generally known in the art and include, for example, inverted abasic residues as well as carbon chains, such as terminal C3H7 (propyl), C6H 13 (Hexyl) or C 12 H 25 (dodecyl) group. In some embodiments, the capping residue is present at either the 5'-end, the 3'-end, or both the 5'-end and the 3'-end of the sense strand. In some embodiments, the 5'-end and / or the 3'-end of the sense strand may contain more than one inverted abasic deoxyribose moiety as a capping residue.

[0074] In some embodiments, one or more inverted abasic residues (invAb) are added to the 3' end of the sense strand. In some embodiments, one or more inverted abasic residues (invAb) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasic residues or inverted abasic sites are inserted between the targeting ligand and the nucleotide sequence of the sense strand of the RNAi agent. In some embodiments, the inclusion of one or more inverted abasic residues or inverted abasic sites at or near the end(s) of the sense strand of the RNAi agent allows for enhanced activity or other desired properties of the RNAi agent.

[0075] In some embodiments, one or more inverted abasic residues (invAb) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasic residues can be inserted between the targeting ligand and the nucleotide sequence of the sense strand of the RNAi agent. The inverted abasic residues can be linked via phosphate, phosphorothioate (e.g., herein designated (invAb)s), or other internucleoside linkages. The chemical structure of an inverted abasic deoxyribose residue is shown in Table A below, and in the chemical structures shown in Figures 1A-1D and 2A-2D.

[0076] [Table A]

[0077] Targeting Moieties and Groups APOC3 RNAi agents may be conjugated to one or more non-nucleotide groups, including but not limited to targeting moieties or targeting groups. Targeting moieties or targeting groups may enhance targeting or delivery of RNAi agents. Examples of targeting moieties and targeting groups are known in the art. Specific examples of (NAG37)s targeting groups used in APOC3 RNAi drug substances described in Table 2 herein, including three N-acetyl-galactosamine targeting moieties, are provided in Table B. Targeting moieties or targeting groups may be covalently linked to the 3'-end and / or 5'-end of either the sense strand and / or the antisense strand. In some embodiments, APOC3 RNAi agents contain targeting groups linked to the 3'-end and / or 5'-end of the sense strand. In some embodiments, the targeting group is linked to the 5'-end of the sense strand of APOC3 RNAi agents. In some embodiments, the targeting group comprises, consists essentially of, or consists of the structure (NAG37)s and is linked to the 5' end of the sense strand of the APOC3 RNAi agent. The targeting group can be linked directly to the RNAi agent or indirectly through a linker / linking group. In some embodiments, the targeting group is linked to the RNAi agent through a labile, cleavable or reversible bond or linker. In some embodiments, the targeting group is linked to an inverted abasic residue at the 5' end of the sense strand.

[0078] Targeting groups or targeting moieties can enhance the pharmacokinetic or biodistribution properties of the conjugate or RNAi agent to which they are attached to improve cell-specific distribution and cell-specific uptake of the conjugate or RNAi agent. In some embodiments, targeting groups enhance the endocytosis of the RNAi agent. Targeting groups can be monovalent, bivalent, trivalent, tetravalent, or have higher valency for the target to which they are directed. Representative targeting groups include, without limitation, compounds with affinity for cell surface molecules, cell receptor ligands, haptens, antibodies, monoclonal antibodies, antibody fragments, and antibody mimics with affinity for cell surface molecules.

[0079] In some embodiments, the targeting group comprises an asialoglycoprotein receptor ligand. In some embodiments, the asialoglycoprotein receptor ligand comprises or consists of one or more galactose derivatives. As used herein, the term galactose derivative includes both galactose and the derivatives of galactose that have affinity for asialoglycoprotein receptors equal to or higher than that of galactose. Galactose derivatives include, but are not limited to, galactose, galactosamine, N-formylgalactosamine, N-acetyl-galactosamine, N-propionyl-galactosamine, Nn-butanoyl-galactosamine and N-iso-butanoyl-galactosamine (see, for example, ST Iobst and K. Drickamer, JBC, 1996, 271, 6686). Galactose derivatives, and clusters of galactose derivatives, useful for in vivo targeting of oligonucleotides and other molecules to the liver are known in the art (see, e.g., Baenziger and Fiete, 1980, Cell, 22, 611-620; Connolly et al., 1982, J. Biol. Chem., 257, 939-945).

[0080] Galactose derivatives have been used to target molecules to hepatocytes in vivo through their binding to asialoglycoprotein receptors expressed on the surface of hepatocytes. Binding of asialoglycoprotein receptor ligands to asialoglycoprotein receptor(s) promotes cell-specific targeting to hepatocytes and endocytosis of molecules into hepatocytes. Asialoglycoprotein receptor ligands can be monomeric (e.g., with a single galactose derivative) or multimeric (e.g., with multiple galactose derivatives). Galactose derivatives or galactose derivative "clusters" can be attached to the 3' or 5' end of the sense or antisense strand of an RNAi agent using methods known in the art.

[0081] In some embodiments, the targeting group comprises a galactose derivative cluster. As used herein, a galactose derivative cluster comprises a molecule having two to four terminal galactose derivatives. The terminal galactose derivative is attached to the molecule through its C-1 carbon. In some embodiments, the galactose derivative cluster is a galactose derivative trimer (also referred to as a tri-antennary galactose derivative or a trivalent galactose derivative). In some embodiments, the galactose derivative cluster comprises an N-acetyl-galactosamine. In some embodiments, the galactose derivative cluster comprises three N-acetyl-galactosamines. In some embodiments, the galactose derivative cluster is a galactose derivative tetramer (also referred to as a tetra-antennary galactose derivative or a tetravalent galactose derivative). In some embodiments, the galactose derivative cluster comprises four N-acetyl-galactosamines.

[0082] As used herein, a galactose derivative trimer contains three galactose derivatives, each linked to a central branch point. As used herein, a galactose derivative tetramer contains four galactose derivatives, each linked to a central branch point. A galactose derivative can be attached to the central branch point via the C-1 carbon of the saccharide. In some embodiments, a galactose derivative is linked to the branch point via a linker or spacer. In some embodiments, the linker or spacer is a flexible hydrophilic spacer, such as a PEG group (see, for example, U.S. Pat. No. 5,885,968; Biessen et al. J. Med. Chem. 1995 Vol. 39 p. 1538-1546). A branch point can be any small molecule that allows the attachment of three galactose derivatives and further allows the attachment of the branch point to an RNAi agent. An example of a branch point group is di-lysine or di-glutamate. The linkage of the branch point to the RNAi agent can occur through a linker or spacer. In some embodiments, the linker or spacer comprises a flexible hydrophilic spacer, for example but not limited to a PEG spacer. In some embodiments, the linker comprises a rigid linker, for example a cyclic group. In some embodiments, the galactose derivative comprises or consists of N-acetyl-galactosamine. In some embodiments, the galactose derivative cluster is composed of galactose derivative tetramers, which can be, for example, N-acetyl-galactosamine tetramers.

[0083] The preparation of galactose derivative clusters containing targeting groups, e.g., N-acetyl-galactosamine, is described, for example, in International Patent Application Publication No. WO2018 / 044350 (Patent Application No. PCT / US2017 / 021147) and International Patent Application Publication No. WO2017 / 156012 (Patent Application No. PCT / US2017 / 021175), the contents of both of which are hereby incorporated by reference in their entireties.

[0084] For example, the targeting ligand conjugated to the APOC3 RNAi agent described in Tables 1A and 1B has the chemical structure (NAG37)s shown in Table B below.

[0085] [Table B]

[0086] APOC3 RNAi Agents and APOC3 RNAi Drug Substances (ADS-005) In some embodiments, an APOC3 RNAi agent used in the methods disclosed herein has the nucleotide sequence of an APOC3 RNAi drug substance (ADS-005) shown in Table 2. The nucleotide sequences of the APOC3 RNAi agents found in APOC3 RNAi drug substances include the antisense strand nucleotide sequences shown in Table 1A below and the sense strand nucleotide sequences shown in Table 1B below.

[0087] [Table 1A]

[0088] [Table 1B]

[0089] As used in Tables 1A, 1B and 2 herein, the following notations are used to denote modified nucleotides, targeting groups and linking groups: A, C, G, I and U denote adenosine, cytidine, guanosine, inosine and uridine, respectively; a, c, g, i and u denote 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf denote 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s denotes a phosphorothioate linkage; (invAb) denotes an inverted abasic deoxyribose residue (see Table A); (NAG37)s denotes the structure shown in Table B above.

[0090] Those skilled in the art will readily understand that unless otherwise indicated by the sequence (e.g., by a phosphorothioate linkage "s"), the monomers, when present in a chain, are linked to one another by 5'-3'-phosphodiester bonds. Those skilled in the art will clearly understand that the inclusion of phosphorothioate linkages as shown in the modified nucleotide sequences disclosed herein replaces the phosphodiester linkages typically present in oligonucleotides (see, e.g., Figures 1-3, which show all linkages). Furthermore, those skilled in the art will readily understand that the terminal nucleotides at the 3' end of a given oligonucleotide sequence, ex vivo, typically have a hydroxyl (-OH) group at the 3' position of each of the given monomers, instead of a phosphate moiety. Furthermore, for the embodiments disclosed herein, when each chain is viewed 5'→3', an inverted abasic residue is inserted such that the 3' position of the deoxyribose is linked at the 3' end of the preceding monomer on each chain. Furthermore, as those skilled in the art will readily understand and appreciate, while the phosphorothioate chemical structures shown herein typically show an anion on the sulfur atom, the invention disclosed herein encompasses all phosphorothioate tautomers (e.g., when the sulfur atom has a double bond and the anion is an oxygen atom).Unless otherwise clearly indicated herein, such understanding of the artisan will be used when describing the APOC3 RNAi agents and compositions comprising APOC3 RNAi agents disclosed herein.

[0091] Each sense and / or antisense strand can have any of the targeting or linking groups listed above, as well as other targeting or linking groups, conjugated to the 5' and / or 3' ends of the sequence.

[0092] The antisense strand sequence of the APOC3 RNAi agent is designed to target mRNA transcripts from the APOC3 gene in a human subject, thereby silencing translation of the APOC3 protein using the RNA interference machinery for a human subject that has APOC3.

[0093] In some embodiments, the methods disclosed herein use an APOC3 RNAi drug substance as shown in Table 2 below:

[0094] [Table 2-1]

[0095] [Table 2-2]

[0096] A schematic representation of the APOC3 RNAi drug substance (ADS-005) is shown in Figure 3, and a complete chemical structure representation is shown in Figures 1A-1D (free acid form) and 2A-2D (sodium salt form). In some embodiments, the APOC3 RNAi drug substance is prepared or provided as a salt, mixed salt or free acid. In some embodiments, the form is the sodium salt.

[0097] Pharmaceutical Compositions and Formulations APOC3 RNAi agents suitable for use in the methods disclosed herein can be prepared as pharmaceutical compositions or formulations for administration to human subjects. Pharmaceutical compositions can be used to treat subjects with diseases or disorders that benefit from the inhibition of APOC3 mRNA expression or the reduction in APOC3 protein levels, such as human subjects with APOC3-related diseases or disorders. In some embodiments, the method comprises administering to a subject in need of treatment an APOC3 RNAi agent linked to a targeting group or targeting ligand as described herein. In some embodiments, one or more pharma- ceutical acceptable excipients, including vehicles, carriers, diluents and / or delivery polymers, are added to the pharmaceutical composition comprising the APOC3 RNAi agent, thereby forming a pharmaceutical formulation suitable for in vivo delivery to human subjects.

[0098] A pharmaceutical composition comprising an APOC3 RNAi agent, when administered to a human subject using the methods disclosed herein, decreases or reduces the level of APOC3 mRNA in the subject.

[0099] In some embodiments, the described pharmaceutical compositions comprising APOC3 RNAi agents are used to treat or manage clinical symptoms in subjects with APOC3-related diseases or disorders.In some embodiments, a therapeutically or prophylactically effective amount of one or more pharmaceutical compositions is administered to a subject in need of such treatment.In some embodiments, administration of any of the disclosed APOC3 RNAi agents can be used to reduce the number, severity and / or frequency of disease symptoms in a subject.

[0100] The described pharmaceutical compositions comprising APOC3 RNAi agents can be used to treat at least one symptom in a subject with a disease or disorder that benefits from the reduction or inhibition of APOC3 protein level expression.In some embodiments, the subject is administered a therapeutically effective amount of one or more of the pharmaceutical compositions comprising APOC3 RNAi agents, thereby treating the symptom.In other embodiments, the subject is administered a prophylactically effective amount of one or more APOC3 RNAi agents, thereby preventing at least one symptom.

[0101] APOC3 RNAi agent disclosed herein can be administered via any suitable route, in a preparation that is appropriately adjusted for a specific route.Therefore, pharmaceutical composition described herein can be administered by injection, for example, intravenously or subcutaneously.In some embodiments, pharmaceutical composition described herein is administered via subcutaneous injection.

[0102] As used herein, a pharmaceutical composition or medicament comprises a pharmacologically effective amount of at least one APOC3 RNAi agent and one or more pharma- ceutically acceptable excipients. A pharma-ceutically acceptable excipient(s) is a substance other than the active ingredient (API, therapeutic product, e.g., APOC3 RNAi agent) that is intentionally included in the drug delivery system. The excipient does not exert or is not intended to exert a therapeutic effect at the intended dosage. The excipient may act a) to aid in the processing of the drug delivery system during manufacture, b) to protect, support or enhance the stability, bioavailability or patient acceptability of the API, c) to aid in product identification, and / or d) to enhance any other aspect of the overall safety, efficacy, or delivery of the API during storage or use. A pharma-ceutically acceptable excipient may or may not be an inert substance.

[0103] Excipients may include, but are not limited to, absorption enhancers, anti-adhesives, anti-foaming agents, antioxidants, binders, buffering agents, carriers, coating agents, pigments, delivery enhancers, delivery polymers, dextran, dextrose, diluents, disintegrants, emulsifiers, bulking agents, fillers, flavoring agents, glidants, humectants, lubricants, oils, polymers, preservatives, saline solutions, salts, solvents, sugars, suspending agents, sustained release matrices, sweeteners, thickeners, tonicity agents, vehicles, water repellents, and wetting agents.

[0104] Suitable pharmaceutical compositions for injectable use include sterile aqueous solutions (if water soluble). For subcutaneous or intravenous administration, suitable carriers may include physiological saline, bacteriostatic water, Cremophor® ELTM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). It must be stable under the conditions of manufacture and storage and should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be, for example, a solvent or dispersion medium containing water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), and suitable mixtures thereof.

[0105] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in a suitable solvent with one or a combination of ingredients listed above, followed by filter sterilization as required. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those listed above.

[0106] In some embodiments, pharmaceutical compositions suitable for use in the methods disclosed herein contain the components identified in the formulated APOC3 RNAi drug substances provided in Table 3 below.

[0107] The APOC3 RNAi agent may be formulated in a composition in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form refers to physically discrete units suitable as unitary dosages for the subjects to be treated; each unit contains a predetermined amount of active compound calculated to produce a desired therapeutic effect in association with the required pharmaceutical carrier. In some embodiments, the dosage unit is between about 1 mg and about 100 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is between about 10 mg and about 100 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is between about 10 mg and about 50 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is about 10 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is about 25 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is about 50 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is about 100 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is about 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96 or is from about 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98 or 100 mg of APOC3 RNAi drug substance.

[0108] The pharmaceutical composition may contain other additional components commonly found in pharmaceutical compositions, including, but not limited to, antipruritic agents, astringents, local anesthetics, or anti-inflammatory agents (e.g., antihistamines, diphenhydramine, etc.).

[0109] As used herein, a "pharmacologically effective amount," "therapeutically effective amount," or simply "effective amount" refers to that amount of an RNAi agent to produce a pharmacological, therapeutic, or prophylactic result.

[0110] The pharma- ceutically acceptable formulations described herein can be packaged in a kit, container, pack, or dispenser.The pharmaceutical compositions described herein can be packaged in pre-filled syringes or vials.

[0111] Formulated APOC3 RNAi drug substances In some embodiments, the APOC3 RNAi drug substance provided in Table 2 (ADS-005) is formulated with one or more pharma- ceutically acceptable excipients to form a pharmaceutical composition suitable for administration to a human subject. In some embodiments, the APOC3 RNAi drug substance described in Table 2 is formulated at 200 mg / mL in aqueous sodium phosphate buffer (0.5 mM sodium dihydrogen phosphate, 0.5 mM disodium hydrogen phosphate) to form the formulated APOC3 RNAi drug substance (ADS-005-1) shown in Table 3:

[0112] [Table 3]

[0113] The APOC3 RNAi drug substance formulated according to Table 3 is prepared as a sterile formulation. In some embodiments, the APOC3 RNAi drug substance formulated is packaged in a container, for example, a glass vial. In some embodiments, the APOC3 RNAi drug substance formulated is packaged in a glass vial with a fill volume of about 1.2mL, and the desired volume for administration can be calculated based on the desired dose level to be administered.

[0114] In some embodiments, a formulated APOC3 RNAi drug substance as shown in Table 3 is administered to a human subject using the methods disclosed herein.

[0115] Human Subjects with Elevated Triglyceride Levels and / or Overexpression of APOC3 The method disclosed herein includes treating a disease, disorder, or other condition or symptom that can be at least partially ameliorated by reducing or silencing APOC3 gene expression in a human subject in need thereof using a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2. In some embodiments, the method disclosed herein includes treating a disease or condition associated with elevated triglycerides in a human subject in need thereof using a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2. In some embodiments, prior to administration, the human subject is diagnosed with an APOC3-related disease or disorder. In some embodiments, the human subject is diagnosed with cardiovascular disease, coronary artery disease, or atherosclerosis. In some embodiments, the human subject has been diagnosed with moderate hypertriglyceridemia (TG levels of 150-499 mg / dL according to the 2018 AHA / ACC Guidelines), severe hypertriglyceridemia (TG>500 mg / dL), and / or chylomicronemia and / or familial chylomicronemia syndrome (FCS) (often TG levels>1000 mg / dL). FCS is a severe rare genetic disease with a prevalence of 1 in 1,000,000, often caused by various single gene mutations (e.g., null mutations in LPL, GPIHBP1, APOC2, APOA5 or LMF1) that result in extremely high TG levels, typically exceeding 900 mg / dL. As shown in the Examples herein, the APOC3 RNAi drug substances described in Table 2 provide substantial TG lowering effects suitable for the treatment of diseases and disorders.

[0116] Medication and inhibition of APOC3 gene expression In general, an effective amount of an APOC3 RNAi agent ranges from about 0.1 to about 10 mg per kg of body weight per dose, for example, from about 0.25 to about 5 mg per kg of body weight per dose. In some embodiments, an effective amount of an APOC3 RNAi agent ranges from about 0.5 to about 4 mg per kg of body weight per dose. In some embodiments, the effective amount is a fixed dose. In some embodiments, a fixed dose of between 1 mg and 100 mg of APOC3 RNAi drug substance is an effective dose. In some embodiments, a fixed dose of between 10 mg and 50 mg of APOC3 RNAi drug substance is an effective dose. In some embodiments, a fixed dose of between 20 mg and 50 mg of APOC3 RNAi drug substance is an effective dose. In some embodiments, a fixed dose of between 25 mg and 50 mg of APOC3 RNAi drug substance is an effective dose. The amount administered is likely to depend on variables such as the patient's overall age and health, the relative biological availability of the compound being delivered, the formulation of the drug, the presence and type of excipients in the formulation, and the route of administration. In some embodiments, a fixed dose of about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg is an effective dose. In some embodiments, a fixed dose of about 10 mg, about 25 mg, about 50 mg, or about 100 mg is an effective dose.

[0117] It is also understood that the initial dosage administered may be increased beyond the upper level in some cases to quickly achieve the desired blood or tissue level, or the initial dosage may be lower than optimal in some cases. For example, in some embodiments, an initial or first dose of about 1 mg to about 100 mg of APOC3 RNAi drug substance is administered, followed by a second dose of about 1 to 100 mg of APOC3 RNAi drug substance approximately one month later, followed by a further dose (similar in concept to a "maintenance dose") once every three months (e.g., once per calendar quarter or once per 12 weeks (q12w)).

[0118] For the treatment of a disease or to form a medicament or composition for the treatment of a disease, the pharmaceutical compositions described herein comprising an APOC3 RNAi agent may be combined with an excipient or with a second therapeutic agent or treatment, including, but not limited to, a second or other RNAi agent, a small molecule drug, an antibody, an antibody fragment, a peptide and / or an aptamer.

[0119] In some embodiments, APOC3 mRNA levels in a subject to which a described APOC3 RNAi agent is administered are reduced in the liver by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or greater than 99%. In some embodiments, the APOC3 mRNA level of a subject administered with a described APOC3 RNAi agent is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, or more than 80% in the entire subject compared to the subject before administration of the APOC3 RNAi agent or the subject not receiving the APOC3 RNAi agent. Gene expression levels and / or mRNA levels in the subject are reduced in cells, groups of cells and / or tissues of the subject.

[0120] In some embodiments, APOC3 protein levels in a subject administered a described APOC3 RNAi agent are reduced in the liver by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or greater than 99% compared to the subject prior to administration of the APOC3 RNAi agent or a subject not receiving the APOC3 RNAi agent. In some embodiments, the APOC3 protein level in a subject administered with the described APOC3 RNAi agent is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, or more than 80% in the whole subject compared to the subject before administration of the APOC3 RNAi agent or the subject not receiving the APOC3 RNAi agent. The protein level in the subject may be reduced in the subject's cells, groups of cells, tissues, blood and / or other bodily fluids.

[0121] The reduction in APOC3 gene expression, APOC3 mRNA or APOC3 protein level can be evaluated and quantified by common methods known in the art.The examples disclosed herein generally show the known methods for evaluating the inhibition of APOC3 gene expression and the reduction in APOC3 protein level.The reduction or reduction in APOC3 mRNA level and / or protein level is collectively referred to herein as the reduction or reduction in APOC3 or inhibiting or reducing the expression of APOC3.

[0122] In some embodiments, the triglyceride level in the subject administered with the described APOC3 RNAi agent is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more than 95% compared to the subject before the APOC3 RNAi agent is administered or the subject who does not receive the APOC3 RNAi agent. The triglyceride level in the subject is reduced in the blood / serum of the subject.

[0123] In some embodiments, the very low density lipoprotein cholesterol (VLDL-C) level in the subject administered with the described APOC3 RNAi agent is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or more than 85% compared to the subject before the APOC3 RNAi agent is administered or the subject who has not received the APOC3 RNAi agent. The VLDL-C level in the subject is reduced in the blood / serum of the subject.

[0124] In some embodiments, the total cholesterol level in a subject administered with the described APOC3 RNAi agent is reduced by at least about 5%, 10%, 15%, 20%, 25%, or more than 25% compared to the subject before administration of the APOC3 RNAi agent or the subject not receiving the APOC3 RNAi agent. The total cholesterol level in the subject is reduced in the subject's cells, groups of cells, tissues, blood and / or other bodily fluids.

[0125] In some embodiments, the high density lipoprotein cholesterol level (HDL-C) in a subject administered a described APOC3 RNAi agent is increased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or more than 85% compared to the subject before administration of the APOC3 RNAi agent or to a subject not receiving the APOC3 RNAi agent. The total HDL-C level in the subject is increased in the subject's cells, groups of cells, tissues, blood and / or other bodily fluids.

[0126] In some embodiments, the apolipoprotein A1 (ApoA1) level in a subject administered with the described APOC3 RNAi agent is increased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more than 95% compared to the subject before the APOC3 RNAi agent is administered or the subject does not receive the APOC3 RNAi agent. The ApoA1 level in the subject is reduced in the subject's cells, groups of cells, tissues, blood and / or other bodily fluids.

[0127] As used herein, the terms "treat", "treatment" and the like refer to a method or step taken to provide relief or a reduction from the number, severity and / or frequency of one or more symptoms of a disease in a subject. As used herein, "treat" and "treatment" can include prevention, management, prophylactic treatment and / or inhibition of the number, severity and / or frequency of one or more symptoms of a disease in a subject. As used herein, unless expressly indicated otherwise, the phrase "treatment of a disease or disorder" includes treatment of the underlying condition and / or symptoms of the disease, as understood by those skilled in the art. Unless expressly limited otherwise herein, such phrases are not intended to be limiting.

[0128] As used herein, "APOC3-related disease or disorder" refers to any disease or disorder that can be treated at least in part by reducing or inhibiting APOC3 gene expression using APOC3 RNAi agents, including by the method of treatment described herein.As discussed herein, the method of treatment disclosed herein, among others, results in a reduction in TG levels in the subject to whom the APOC3 RNAi drug substance is administered.APOC3-related diseases and disorders include, but are not limited to, familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, lipodystrophy syndrome including familial partial lipodystrophy, obesity, dyslipidemia, nonalcoholic steatohepatitis, nonalcoholic fatty liver disease, hyperlipidemia, hypertriglyceridemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, and other dyslipidemia and metabolic-related disorders and diseases.

[0129] As used herein, "monthly dosing" or "monthly" dosing means every 28 days. As used herein, "quarterly dosing" or "quarterly" dosing means every 84 days. When used in conjunction with monthly dosing, the term "about" means + / - 5 days of monthly dosing. When used in conjunction with quarterly dosing, the term "about" means + / - 14 days of quarterly dosing.

[0130] As used herein, the phrase "introduce into cell" refers to RNAi agent, and means to functionally deliver RNAi agent into cell.The phrase "functionally deliver" means to deliver RNAi agent into cell in such a manner that allows RNAi agent to have expected biological activity, for example, sequence-specific inhibition of gene expression.

[0131] Unless otherwise stated, the symbols [ka] The use of as used herein means that any group or groups according to the scope of the invention described herein may be linked thereto.

[0132] As used herein, unless specifically identified in a structure having a particular conformation, for each structure in which asymmetric centers exist and thus give rise to enantiomers, diastereomers or other stereoisomeric configurations, each structure disclosed herein is intended to represent all such possible isomers, including their optically pure and racemic forms. For example, the structures disclosed herein are intended to cover mixtures of diastereomers as well as single stereoisomers.

[0133] As used in the claims herein, the phrase "consisting of" excludes any element, step, or ingredient not specified in the claim. As used in the claims herein, the phrase "consisting essentially of" limits the claim to those materials or steps specified and that do not materially affect the basic and novel feature(s) of the claimed invention.

[0134] Those skilled in the art will readily understand and appreciate that the compounds and compositions disclosed herein may have certain atoms (e.g., N, O, or S atoms) in a protonated or deprotonated state depending on the environment in which the compound or composition is placed. Thus, as used herein, the structures disclosed herein assume that certain functional groups, such as OH, SH, or NH, may be protonated or deprotonated. The disclosure of the present specification is intended to cover the disclosed compounds and compositions regardless of the state of protonation based on the environment (e.g., pH), as will be readily understood by those skilled in the art.

[0135] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.Methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, but suitable methods and materials are described below.All publications, patent applications, patents and other references mentioned herein are hereby incorporated by reference in their entirety.In case of conflict, the present specification, including definitions, will control.In addition, materials, methods and examples are illustrative only and are not intended to be limiting.

[0136] The embodiments and provisions provided above will now be illustrated with the following non-limiting examples. EXAMPLES

[0137] Example 1 Synthesis and formulation of APOC3 RNAi drug substance (ADS-005) APOC3 RNAi drug substances suitable for use in the methods disclosed herein can be synthesized using standard phosphoramidite techniques in solid-phase oligonucleotide synthesis, as known in the art. Commercially available oligonucleotide synthesizers (e.g., MerMade96E® (Bioautomation) or MerMade12® (Bioautomation)) can be used. Synthesis can be carried out on solid supports made of controlled pore glass (CPG, 500 Å or 600 Å, obtained from Prime Synthesis, Aston, PA, USA). The monomers located at the 3' end of each strand can be attached to the solid support as starting points for synthesis. All RNA, 2'-modified RNA phosphoramidites and inverted abasic phosphoramidites can be purchased commercially. A targeting group-containing phosphoramidite suitable for addition to the 5' end of the sense strand can be synthesized. Standard cleavage, deprotection, purification and annealing steps can be utilized, as known in the art. Further description of the synthesis of APOC3 RNAi agents can be found, for example, in International Patent Application Publication No. WO2019 / 051402 (Patent Application No. PCT / US2018 / 050248) and WO2018 / 044350 (PCT / US2017 / 021147), each of which is hereby incorporated by reference in its entirety.The APOC3 RNAi drug substance can then be formulated by dissolving in a standard pharma- ceutically acceptable excipient generally known in the art.For example, Table 3 shows formulated APOC3 RNAi drug substances suitable for use in the methods disclosed herein.

[0138] Example 2 Phase I clinical trial of an APOC3 RNAi drug substance (ADS-005) in healthy human volunteers and in patients with severe hypertriglyceridemia and familial chylomicronemia syndrome (FCS) A Phase 1 single and multiple ascending dose study was undertaken to evaluate the safety, tolerability, pharmacokinetic and pharmacodynamic effects of an APOC3 RNAi drug substance (ADS-005) in healthy adult volunteers and in patients with severe hypertriglyceridemia and FCS. The study subject population consisted of subjects aged 19.0 kg / m 2 and 40.0 kg / m 2 Includes adult men and women aged 18-65 years with a BMI between Cohorts 1, 2, 3 and 4: All subjects have fasting screening triglycerides >80 mg / dL (>0.903 mmol / L) and are not receiving any lipid- or triglyceride-lowering treatment. Each double-blind cohort will enroll 10 subjects (6 active drug: 4 placebo (PBO)) and all cohorts will receive a single ascending dose of APOC3 RNAi drug substance or PBO at ascending dose levels of 10, 25, 50 and 100 mg as per Figures 4 and 5. Cohorts 1b, 2b, 3b and 4b: Cohorts will be double-blinded with 8 active drug and 2 placebo patients per cohort with fasting serum triglycerides of at least 300 mg / dL (3.38 mmol / L) at screening. All subjects will receive multiple ascending doses of APOC3 RNAi drug substance or placebo at dose levels of 10, 25, 50 or 100 mg. Cohort 5: The cohort will include up to eight chylomicronemia patients and will be open-label. Subjects in Cohort 5 will each receive two 50 mg doses of the APOC3 RNAi drug substance. Cohorts 6, 7, and 8: All subjects have fasting screening triglycerides >80 mg / dL (>0.903 mmol / L) and are not receiving any lipid- or triglyceride-lowering treatment. Each double-blind cohort will enroll 10 subjects (6 active drug: 4 placebo (PBO)) and all cohorts will receive a single ascending dose of APOC3 RNAi drug substance or PBO at ascending dose levels of 10, 25, 50, and 100 mg as per Figures 4 and 5.

[0139] Subjects are separated into different cohorts, taking into account the criteria discussed above (see also Figure 5). Cohorts 1 to 4 are randomized to receive APOC3 RNAi drug substance or placebo in a single dose of 25 mg (cohort 1), 50 mg (cohort 2), 100 mg (cohort 3) or 10 mg (cohort 4), administered as a subcutaneous injection (6 active drug: 4 placebo). In the other cohorts, two doses (administered on days 1 and 29) of 10 mg (cohort 1b and cohort 6), 50 mg (cohort 2b and cohort 8), 100 mg (cohort 3b) or 25 mg (cohort 4b and cohort 7) are administered via subcutaneous injection. In addition, two doses of 50 mg are administered to chylomicronemia patients (cohort 5). Cohorts 1, 2, 3, 4, 1b, 2b, 3b and 4b will be double-blinded. Cohorts 5, 6, 7 and 8 will be open-label. A total of up to 80 subjects may be enrolled in the study. Figure 4 shows the study design for the Phase I clinical trial, revised. Study parameters are summarized in Table 4 below.

[0140] [Table 4-1] [Table 4-2] [Table 4-3]

[0141] Initially, a cohort was proposed as part of the clinical trial protocol with 200 mg of APOC3 RNAi drug substance per dose. However, in light of the unexpected and surprising efficacy at the 25 mg and 50 mg doses, the study protocol was revised to remove the 200 mg cohort. The 200 mg cohort was replaced with a cohort dosing at a lower dose of 10 mg. The 10 mg, 25 mg, 50 mg and 100 mg doses all produced substantial serum APOC3 reduction. For example, both the 25 mg and 50 mg cohorts reached approximately 85% mean serum APOC3 protein reduction after a single dose in the Phase I study by day 22, and the 100 mg cohort reached approximately 85% mean serum APOC3 protein reduction after a single dose in the Phase I study by day 8. Figures 6-9 report on the serum APOC3 reduction for cohorts 1, 2, 3 and 4 in the Phase I study.

[0142] The serum APOC3 reduction results from the study show that administration of APOC3 RNAi drug substance from at least 10mg, 25mg, 50mg and 100mg doses produced a deep and sustained reduction in serum APOC3 when compared to placebo.As pointed out herein, APOC3 is mainly synthesized in hepatocytes in the liver, but data suggests that as much as 20% of APOC3 protein may be expressed in enterocytes in the gastrointestinal tract.Surprisingly and unexpectedly, a dose level of only 25mg APOC3 RNAi drug substance produced substantial knockdown of APOC3 gene expression, reaching approximately 85% (which was a similar knockdown level observed at the higher doses of 50mg and 100mg), indicating the achievement of nearly complete suppression of APOC3 expressed in hepatocytes.Also unexpectedly, a dose level of only 10mg APOC3 RNAi drug substance produced substantial knockdown, showing a maximum reduction of approximately 72% in APOC3 gene expression compared to baseline values.

[0143] The duration of serum APOC3 reduction (>80%) from a single dose of 25 mg also appears to be unexpectedly long-lasting. By 85 days after dose administration, ≧70% APOC3 reduction was observed at day 113 (end of study). Even at the 10 mg dose, the duration of effect was long, with >60% APOC3 reduction observed by 57 days after dose administration. These data indicate that monthly, quarterly, or even once every 6 months dosing with 10 mg, 25 mg, 50 mg, or 100 mg can achieve substantial APOC3 gene expression inhibition.

[0144] Substantial reductions in serum triglyceride (TG) levels were also observed across the various cohorts after administration of only a single dose. Figures 9-12 report on serum TG reductions in cohorts 1, 2, 3 and 4 in the Phase I study. Each of the 25 mg (cohort 1), 50 mg (cohort 2) and 100 mg (cohort 3) dosing groups reached a maximum TG reduction of approximately 60% compared to baseline. By day 113 (end of study), both the 25 mg (cohort 1) and 50 mg (cohort 2) maintained a reduction of approximately 50% in serum TG. Additionally, the 10 mg (cohort 4) dosing group reached a maximum TG reduction of approximately 50% and maintained a reduction of approximately 40% in serum TG at day 99.

[0145] Reductions in VLDL-C similar to those of serum TG were also observed. For example, at day 29, the 25 mg (cohort 1), 50 mg (cohort 2) and 100 mg (cohort 3) dosing groups each achieved approximately 60% reduction in VLDL-C levels, and at day 29, the 10 mg dosing group (cohort 4) reported an approximately 50% reduction. Additionally, the data show approximately 10% to 20% reduction in total cholesterol and low-density lipoprotein cholesterol (LDL-C) in subjects.

[0146] Additionally, administration of APOC3 RNAi drug substances at the dose levels disclosed herein has shown approximately 50%-70% increases in high density lipoprotein cholesterol (HDL-C) and apolipoprotein A1 (APOA1) levels in Phase 1 studies, from which it has been observed that the HDL-C increases are slightly greater than those seen in APOC3 deficient individuals (homozygous or heterozygous).

[0147] Multiple dose data from patients in cohort 5 (n=5, all chylomicronemia patients with data through day 29) also showed reductions in serum APOC3 protein levels and lipid parameters. For APOC3 protein levels, a maximum mean reduction of 96% (-62 mg / dL) was observed across three subjects, with a mean baseline APOC3 level of 66 mg / dL. In these five subjects, a maximum mean reduction of 92% (-2583 mg / dL) was observed in triglycerides, with a mean baseline value of 2787 mg / dL ± 1444 mg / dL (standard deviation). In one of the subjects, a maximum individual reduction of 99% (-86.7 mg / dL) was observed after a single dose, with a starting concentration of 88 mg / dL. Additionally, a maximum individual reduction of 95% (-4410 mg / dL) was observed in one subject 28 days after the first dose, who had a starting triglyceride value of 4636 mg / dL. Figure 26 shows APOC3 protein levels from cohort 5, and Figure 27 shows triglyceride levels from cohort 5.

[0148] To date, there have been no deaths, no serious adverse events (SAEs), and no adverse events (AEs) rated as severe in intensity in the Phase I study. Most AEs have been reported as mild.

[0149] Other embodiments In some embodiments, disclosed herein is a method for treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising a first dose of the APOC3 RNAi agent of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi agent about one month after the first dose, wherein the first dose and second dose are administered by subcutaneous injection.

[0150] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising a first dose of the APOC3 RNAi agent of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi agent about three months after the first dose (e.g., quarterly dosing or every 12 weeks dosing (q12w)), wherein the first dose and second dose are administered by subcutaneous injection.

[0151] In some embodiments, disclosed herein is a method for treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising a first dose of the APOC3 RNAi agent of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi agent about four months after the first dose, wherein the first dose and second dose are administered by subcutaneous injection.

[0152] In some embodiments, disclosed herein is a method for treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising a first dose of the APOC3 RNAi agent of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi agent about 6 months after the first dose, wherein the first dose and second dose are administered by subcutaneous injection.

[0153] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject a first dose of a pharmaceutical composition comprising an APOC3 RNAi agent at a dose of between about 1 mg and about 100 mg of APOC3 RNAi drug substance by subcutaneous administration; b. administering to the human subject a second dose of the pharmaceutical composition about one month after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 3 to about 6 months after the second dose. Disclosed herein is a method comprising:

[0154] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject a first dose of a pharmaceutical composition comprising an APOC3 RNAi agent by subcutaneous administration, the pharmaceutical composition comprising an APOC3 RNAi agent at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent; b. administering to the human subject a second dose of the pharmaceutical composition about one month after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 6 months after the second dose. Disclosed herein is a method comprising:

[0155] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject a first dose of a pharmaceutical composition comprising an APOC3 RNAi agent by subcutaneous administration, the pharmaceutical composition comprising an APOC3 RNAi agent at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent; b. administering to the human subject a second dose of the pharmaceutical composition about 1 month to about 6 months after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 3 months (e.g., quarterly dosing or every 12 weeks (q12w) dosing) to about 6 months after the second dose. Disclosed herein is a method comprising:

[0156] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject a first dose of a pharmaceutical composition comprising an APOC3 RNAi agent at a dose of between about 1 mg and about 100 mg of APOC3 RNAi drug substance by subcutaneous administration; b. administering to the human subject a second dose of the pharmaceutical composition about 3 months to about 6 months after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 6 months after the second dose. Disclosed herein is a method comprising:

[0157] In some embodiments, in the methods disclosed herein, each dose is between about 10 mg and about 100 mg of an APOC3 RNAi agent.

[0158] In some embodiments, in the methods disclosed herein, each dose is between about 10 mg and about 50 mg of an APOC3 RNAi agent.

[0159] In some embodiments, in the methods disclosed herein, each dose is between about 25 mg and about 100 mg of an APOC3 RNAi agent.

[0160] In some embodiments, in the methods disclosed herein, each dose is between about 25 mg and about 50 mg of an APOC3 RNAi agent.

[0161] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi agent is about 10 mg.

[0162] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi agent is about 25 mg.

[0163] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi agent is about 50 mg.

[0164] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi agent is about 100 mg.

[0165] In some embodiments, in the methods disclosed herein, the subject is further administered an additional therapeutic agent for the treatment of the APOC3-related disease or disorder.

[0166] In some embodiments, in the methods disclosed herein, the APOC3-related disease or disorder is dyslipidemia.

[0167] In some embodiments, in the methods disclosed herein, the APOC3-related disease or disorder is hypertriglyceridemia, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, or familial partial lipodystrophy.

[0168] In some embodiments, in the methods disclosed herein, the APOC3-related disease or disorder is familial chylomicronemia syndrome (FCS), chylomicronemia, or multifactorial chylomicronemia.

[0169] In some embodiments, in the methods disclosed herein, the APOC3-related disease or disorder is hypertriglyceridemia, either with or without a history of pancreatitis.

[0170] In some embodiments, in the methods disclosed herein, the pharmaceutical compositions are packaged in a kit, container, pack, dispenser, pre-filled syringe or vial.

[0171] In some embodiments, disclosed herein is a method for treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at an initial dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of a pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance described in Table 2 about one month after the initial dose, wherein the initial dose and second dose are administered by subcutaneous injection.

[0172] In some embodiments, disclosed herein is a method for treating an APOC3-associated disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at an initial dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of a pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance described in Table 2 about three months after the initial dose (e.g., quarterly dosing or every 12 weeks dosing (q12w)), wherein the initial dose and second dose are administered by subcutaneous injection.

[0173] In some embodiments, disclosed herein is a method for treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at an initial dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of a pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance described in Table 2 about four months after the initial dose, wherein the initial dose and second dose are administered by subcutaneous injection.

[0174] In some embodiments, disclosed herein is a method for treating an APOC3 associated disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at an initial dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of a pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance described in Table 2 about 6 months after the initial dose, wherein the initial dose and second dose are administered by subcutaneous injection.

[0175] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject an initial dose of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance by subcutaneous administration; b. administering to the human subject a second dose of the pharmaceutical composition about one month after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 3 to about 6 months after the second dose. Disclosed herein is a method comprising:

[0176] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject an initial dose of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance by subcutaneous administration; b. administering to the human subject a second dose of the pharmaceutical composition about one month after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 6 months after the second dose. Disclosed herein is a method comprising:

[0177] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject an initial dose of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance by subcutaneous administration; b. administering to the human subject a second dose of the pharmaceutical composition about 1 month to about 6 months after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 3 months (e.g., quarterly dosing or every 12 weeks (q12w) dosing) to about 6 months after the second dose. Disclosed herein is a method comprising:

[0178] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject an initial dose of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance by subcutaneous administration; b. administering to the human subject a second dose of the pharmaceutical composition about 3 months to about 6 months after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 6 months after the second dose. Disclosed herein is a method comprising:

[0179] In some embodiments, in the methods disclosed herein, each dose is between about 10 mg and about 100 mg of the APOC3 RNAi drug substance.

[0180] In some embodiments, in the methods disclosed herein, each dose is between about 10 mg and about 50 mg of the APOC3 RNAi drug substance.

[0181] In some embodiments, in the methods disclosed herein, each dose is between about 25 mg and about 100 mg of the APOC3 RNAi drug substance.

[0182] In some embodiments, in the methods disclosed herein, each dose is between about 25 mg and about 50 mg of the APOC3 RNAi drug substance.

[0183] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi drug substance is about 10 mg.

[0184] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi drug substance is about 25 mg.

[0185] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi drug substance is about 50 mg.

[0186] In some embodiments, in the methods disclosed herein, each dose of the APOC3 RNAi drug substance is about 100 mg.

[0187] In some embodiments, in the methods disclosed herein, the pharmaceutical composition comprises, consists of, or consists essentially of a formulated APOC3 RNAi drug substance as described in Table 3.

[0188] In some embodiments, the methods disclosed herein further comprise administering additional doses after the first dose, where the additional doses are administered about 1 month apart, about 3 months apart (e.g., 12 weeks apart or once per calendar quarter), about 4 months apart, or about 6 months apart.

[0189] In some embodiments, the methods disclosed herein further comprise administering additional doses after the second dose, wherein the additional doses are administered about 1 month apart, about 3 months apart (e.g., 12 weeks apart or once per calendar quarter), about 4 months apart, or about 6 months apart.

[0190] In some embodiments, the methods disclosed herein further comprise administering additional doses after the third dose, wherein the additional doses are administered about 1 month apart, about 3 months apart (e.g., 12 weeks apart or once per calendar quarter), about 4 months apart, or about 6 months apart.

[0191] In some embodiments, the methods disclosed herein further comprise administering an additional dose about 3 to about 6 months apart.

[0192] In some embodiments, disclosed herein is a method of inhibiting expression of APOC3 in a subject in need thereof, comprising administering to the subject an APOC3 RNAi drug substance described in Table 2, wherein the subject is administered one or more doses of the APOC3 RNAi drug substance described in Table 2 in an amount of from about 1 mg to about 100 mg over the course of about one month.

[0193] In some embodiments, disclosed herein is a method of inhibiting expression of APOC3 in a subject in need thereof, comprising administering to the subject an APOC3 RNAi drug substance described in Table 2, wherein the subject is administered one or more doses of the APOC3 RNAi drug substance described in Table 2 in an amount of from about 1 mg to about 100 mg over the course of about 3 months.

[0194] In some embodiments, in the methods disclosed herein, the dose is between about 10 mg and about 100 mg of the APOC3 RNAi drug substance.

[0195] In some embodiments, in the methods disclosed herein, the dose is between about 10 mg and about 50 mg of the APOC3 RNAi drug substance.

[0196] In some embodiments, in the methods disclosed herein, the dose is between about 25 mg and about 100 mg of the APOC3 RNAi drug substance.

[0197] In some embodiments, in the methods disclosed herein, the dose is between about 25 mg and about 50 mg of the APOC3 RNAi drug substance.

[0198] In some embodiments, in the methods disclosed herein, the dose is about 10 mg of the APOC3 RNAi drug substance.

[0199] In some embodiments, in the methods disclosed herein, the dose is about 25 mg of the APOC3 RNAi drug substance.

[0200] In some embodiments, in the methods disclosed herein, the dose is about 50 mg of the APOC3 RNAi drug substance.

[0201] In some embodiments, in the methods disclosed herein, the dose is about 100 mg of the APOC3 RNAi drug substance.

[0202] In some embodiments, a pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance as set forth in Table 2, preferably in a pharma- ceutical acceptable salt form, wherein the APOC3 RNAi drug substance is an APOC3 RNAi drug substance. The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the pharmaceutical composition is administered to the subject at an initial dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance, and about one month after the initial dose, the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance set forth in Table 2, wherein the initial dose and second dose are administered by subcutaneous injection.

[0203] In some embodiments, a pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance as set forth in Table 2, preferably in a pharma- ceutical acceptable salt form, wherein the APOC3 RNAi drug substance is an APOC3 RNAi drug substance. The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the pharmaceutical composition is administered to the subject at an initial dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance, and about 3 months after the initial dose, the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance set forth in Table 2, wherein the initial dose and second dose are administered by subcutaneous injection.

[0204] In some embodiments, a pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance as set forth in Table 2, preferably in a pharma- ceutical acceptable salt form, wherein the APOC3 RNAi drug substance is an APOC3 RNAi drug substance. The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the pharmaceutical composition is administered to the subject at an initial dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance, and about 4 months after the initial dose, the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance set forth in Table 2, wherein the initial dose and second dose are administered by subcutaneous injection.

[0205] In some embodiments, a pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance as set forth in Table 2, preferably in a pharma- ceutical acceptable salt form, wherein the APOC3 RNAi drug substance is an APOC3 RNAi drug substance. The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the pharmaceutical composition is administered to the subject at an initial dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance, and about 6 months after the initial dose, the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance set forth in Table 2, wherein the initial dose and second dose are administered by subcutaneous injection.

[0206] In some embodiments, a pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administering a pharmaceutical composition comprising an APOC3 RNAi drug substance as described in Table 2, a. the pharmaceutical composition is administered to a human subject at an initial dose of between about 1 mg and about 100 mg of APOC3 RNAi drug substance; b. the pharmaceutical composition is administered to a human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about one month after the first dose; c. The pharmaceutical composition is administered to a human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 3 months to about 6 months after the second dose. Pharmaceutical compositions are disclosed herein.

[0207] In some embodiments, a pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administering a pharmaceutical composition comprising an APOC3 RNAi drug substance as described in Table 2, a. the pharmaceutical composition is administered to a human subject at an initial dose of between about 1 mg and about 100 mg of APOC3 RNAi drug substance; b. the pharmaceutical composition is administered to a human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about one month after the first dose; c. The pharmaceutical composition is administered to a human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 6 months after the second dose. Pharmaceutical compositions are disclosed herein.

[0208] In some embodiments, a pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administering a pharmaceutical composition comprising an APOC3 RNAi drug substance as described in Table 2, a. the pharmaceutical composition is administered to a human subject at an initial dose of between about 1 mg and about 100 mg of APOC3 RNAi drug substance; b. the pharmaceutical composition is administered to a human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 1 month to about 6 months after the first dose; c. The pharmaceutical composition is administered to a human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 3 months to about 6 months after the second dose. Pharmaceutical compositions are disclosed herein.

[0209] A pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need of such treatment, said use comprising administering a pharmaceutical composition comprising an APOC3 RNAi drug substance as described in Table 2; a. the pharmaceutical composition is administered to a human subject at an initial dose of between about 1 mg and about 100 mg of APOC3 RNAi drug substance; b. the pharmaceutical composition is administered to a human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 3 months to about 6 months after the first dose; c. The pharmaceutical composition is administered to a human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 6 months after the second dose. Pharmaceutical compositions.

[0210] In some embodiments, for pharmaceutical compositions when used as disclosed herein, each dose of an APOC3 RNAi drug substance is between about 10 mg and about 100 mg.

[0211] In some embodiments, for a pharmaceutical composition when used as disclosed herein, each dose is between about 10 mg and about 50 mg of the APOC3 RNAi drug substance.

[0212] In some embodiments, for a pharmaceutical composition when used as disclosed herein, each dose is between about 25 mg and about 100 mg of the APOC3 RNAi drug substance.

[0213] In some embodiments, for a pharmaceutical composition when used as disclosed herein, each dose is between about 25 mg and about 50 mg of the APOC3 RNAi drug substance.

[0214] In some embodiments, for a pharmaceutical composition when used as disclosed herein, each dose is about 10 mg of APOC3 RNAi drug substance.

[0215] In some embodiments, for a pharmaceutical composition when used as disclosed herein, each dose is about 25 mg of APOC3 RNAi drug substance.

[0216] In some embodiments, for a pharmaceutical composition when used as disclosed herein, each dose is about 50 mg of APOC3 RNAi drug substance.

[0217] In some embodiments, for a pharmaceutical composition when used as disclosed herein, each dose is about 100 mg of the APOC3 RNAi drug substance.

[0218] In some embodiments, the pharmaceutical composition when used as disclosed herein is administered to a human subject along with an additional therapeutic agent for the treatment of an APOC3-related disease or disorder.

[0219] In some embodiments, the pharmaceutical compositions when used as disclosed herein are used to treat dyslipidemia.

[0220] In some embodiments, the pharmaceutical compositions when used as disclosed herein are used to treat hypertriglyceridemia, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, or familial partial lipodystrophy.

[0221] In some embodiments, the pharmaceutical compositions when used as disclosed herein are used to treat familial chylomicronemia syndrome (FCS), chylomicronemia or multifactorial chylomicronemia.

[0222] In some embodiments, the pharmaceutical compositions when used as disclosed herein are used to treat hypertriglyceridemia with or without a history of pancreatitis.

[0223] In some embodiments, the pharmaceutical compositions when used as disclosed herein are packaged in a kit, container, pack, dispenser, pre-filled syringe or vial.

[0224] In some embodiments, the pharmaceutical compositions when used as disclosed herein comprise, consist of, or consist essentially of a formulated APOC3 RNAi drug substance as described in Table 3.

[0225] In some embodiments, the pharmaceutical compositions when used as disclosed herein are administered to the subject with additional doses administered about one month apart.

[0226] In some embodiments, the pharmaceutical composition when used as disclosed herein is administered to the subject with additional doses administered about 3 months apart.

[0227] In some embodiments, the pharmaceutical composition when used as disclosed herein is administered to the subject with additional doses administered about four months apart.

[0228] In some embodiments, the pharmaceutical composition when used as disclosed herein is administered to the subject with additional doses administered about 6 months apart.

[0229] In some embodiments, the pharmaceutical compositions when used as disclosed herein are administered to the subject with additional doses administered about 3 to about 6 months apart.

[0230] In some embodiments, the pharmaceutical compositions when used as disclosed herein are self-administered by the subject.

[0231] In some embodiments, the pharmaceutical compositions when used as disclosed herein are self-administered by a medical professional.

[0232] In some embodiments, the pharmaceutical composition when used as disclosed herein reduces serum APOC3 protein levels by more than 60% compared to baseline levels in a human subject.

[0233] In some embodiments, the pharmaceutical compositions when used as disclosed herein reduce serum triglyceride (TG) levels by more than 50% compared to baseline levels in a human subject.

[0234] In some embodiments, the pharmaceutical compositions when used as disclosed herein reduce serum triglyceride (TG) levels by more than 75% in a human subject.

[0235] In some embodiments, the pharmaceutical compositions when used as disclosed herein reduce very low density lipoprotein cholesterol (VLDL-C) levels, serum low density lipoprotein cholesterol (LDL-C) levels, or both serum VLDL-C and serum LDL-C levels in a human subject compared to baseline levels.

[0236] In some embodiments, a pharmaceutical composition when used as disclosed herein is comprised of an APOC3 RNAi drug substance in the form of a salt, mixed salt, free acid, or combinations thereof.

[0237] In some embodiments, the pharmaceutical compositions when used as disclosed herein are comprised of an APOC3 RNAi drug substance in the form of a pharma- ceutically acceptable sodium salt.

[0238] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of an APOC3 RNAi drug substance described in Table 2 in an amount of from about 1 mg to about 100 mg over the course of about one month.

[0239] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of the APOC3 RNAi drug substance described in Table 2 in an amount of from about 1 mg to about 100 mg over the course of about 3 months.

[0240] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of an APOC3 RNAi drug substance described in Table 2 in an amount of about 10 mg to about 100 mg over the course of about one month.

[0241] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of the APOC3 RNAi drug substance described in Table 2 in an amount of about 10 mg to about 100 mg over the course of about 3 months.

[0242] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of the APOC3 RNAi drug substance described in Table 2 in an amount of about 10 mg to about 100 mg over the course of about 6 months.

[0243] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of an APOC3 RNAi drug substance described in Table 2 in an amount of about 25 mg to about 50 mg over the course of about one month.

[0244] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of an APOC3 RNAi drug substance described in Table 2 in an amount of about 25 mg to about 50 mg over the course of about three months.

[0245] In some embodiments, disclosed herein is a pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2, wherein the pharmaceutical composition is administered in one or more doses of an APOC3 RNAi drug substance described in Table 2 in an amount of about 25 mg to about 50 mg over the course of about 6 months.

[0246] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 100 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least one month elapses between doses, and wherein the doses are administered by subcutaneous injection.

[0247] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 100 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least about 12 weeks elapse between doses (q12w dosing), and wherein the doses are administered by subcutaneous injection.

[0248] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 100 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least about 3 months lapse between doses, and wherein the doses are administered by subcutaneous injection.

[0249] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 100 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least about 6 months elapse between doses, and wherein the doses are administered by subcutaneous injection.

[0250] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 50 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least one month elapses between doses, and wherein the doses are administered by subcutaneous injection.

[0251] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 50 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least about 12 weeks elapse between doses (q12w dosing), and wherein the doses are administered by subcutaneous injection.

[0252] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 50 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least about 3 months lapse between doses, and wherein the doses are administered by subcutaneous injection.

[0253] In some embodiments, disclosed herein is a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance described in Table 2 at a dose of about 50 mg or less of the APOC3 RNAi drug substance, and administering a subsequent dose to the subject, wherein at least about 6 months elapse between doses, and wherein the doses are administered by subcutaneous injection.

[0254] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject a dose of about 100 mg or less of a pharmaceutical composition comprising an APOC3 RNAi drug substance; b. administering to the human subject a second dose of the pharmaceutical composition about 1 month to about 6 months after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 6 months after the second dose. and wherein the dose is administered by subcutaneous administration.

[0255] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject a dose of about 100 mg or less of a pharmaceutical composition comprising an APOC3 RNAi drug substance; b. administering to the human subject a second dose of the pharmaceutical composition about one month after the first dose; and c. Disclosed herein are methods comprising administering to a human subject a third dose of the pharmaceutical composition about 3 months (e.g., every calendar quarter, or once every 12 weeks (q12w)) to about 6 months after the second dose, wherein the doses are administered by subcutaneous administration.

[0256] In some embodiments, there is provided a method for treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising: a. administering to a human subject a dose of about 100 mg or less of a pharmaceutical composition comprising an APOC3 RNAi drug substance; b. administering to the human subject a second dose of the pharmaceutical composition about 3 months (e.g., once every calendar quarter or once every 12 weeks (q12w)) to about 6 months after the first dose; and c. administering to the human subject a third dose of the pharmaceutical composition about 6 months after the second dose. and wherein the dose is administered by subcutaneous administration.

[0257] Although the present invention has been described in conjunction with its detailed description, it should be understood that the above description is intended to illustrate, but not to limit, the scope of the invention as defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. In certain embodiments, for example, the following items are provided: (Item 1) A method of treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the APOC3 RNAi drug substance is administered in a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about one month after the first dose, wherein the first dose and the second dose are administered by subcutaneous injection. (Item 2) A method of treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the APOC3 RNAi drug substance is administered in a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 3 months after the first dose, wherein the first dose and the second dose are administered by subcutaneous injection. (Item 3) A method of treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the APOC3 RNAi drug substance is administered in a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 4 months after the first dose, wherein the first dose and the second dose are administered by subcutaneous injection. (Item 4) A method of treating an APOC3-related disease or disorder in a human subject in need of such treatment, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, comprising administering to said subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the APOC3 RNAi drug substance is administered in a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance, and administering to the subject a second dose of the pharmaceutical composition comprising between about 1 mg and about 100 mg of the APOC3 RNAi drug substance about 6 months after the first dose, wherein the first dose and the second dose are administered by subcutaneous injection. (Item 5) 5. The method of any one of items 1 to 4, further comprising administering an additional dose after the second dose, wherein the additional doses are administered about one month apart. (Item 6) 5. The method of any one of items 1 to 4, further comprising administering an additional dose after the second dose, wherein the additional doses are administered about 3 months apart. (Item 7) 5. The method of any one of items 1 to 4, further comprising administering an additional dose after the second dose, wherein the additional doses are administered about 4 months apart. (Item 8) 5. The method of any one of items 1 to 4, further comprising administering an additional dose after the second dose, wherein the additional doses are administered about 6 months apart. (Item 9) 1. A method of treating an APOC3-related disease or disorder in a human subject in need thereof, comprising: a. administering to said human subject a first dose of a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharma- ceutical acceptable salt form, said APOC3 The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] wherein the APOC3 RNAi drug substance is administered at a dose of between about 1 mg and about 100 mg of the APOC3 RNAi drug substance; b. administering to the human subject a second dose of the pharmaceutical composition about 1 month to about 6 months after the first dose; and c. administering to said human subject a third dose of said pharmaceutical composition about 3 months to about 6 months after said second dose. wherein the first dose, the second dose and the third dose are each administered by subcutaneous injection. (Item 10) 6. The method of claim 5, wherein the second dose is administered about one month after the first dose. (Item 11) 6. The method of claim 5, wherein the second dose is administered about 3 months after the first dose. (Item 12) 6. The method of claim 5, wherein the second dose is administered about 4 months after the first dose. (Item 13) 6. The method of claim 5, wherein the second dose is administered about 6 months after the first dose. (Item 14) 14. The method of any one of items 5 to 13, wherein the third dose is administered about 3 months after the second dose. (Item 15) 14. The method of any one of items 5 to 13, wherein the third dose is administered about 4 months after the second dose. (Item 16) 14. The method of any one of items 5 to 13, wherein the third dose is administered about 6 months after the second dose. (Item 17) 14. The method of any one of items 5 to 13, further comprising administering an additional dose after the third dose, wherein the additional dose is administered about 1 month to about 6 months apart. (Item 18) 14. The method of any one of items 5 to 13, further comprising administering an additional dose after the third dose, wherein the additional dose is administered about 3 months to about 6 months apart. (Item 19) 14. The method of any one of items 5 to 13, further comprising administering an additional dose after the third dose, wherein the additional doses are administered about 6 months apart. (Item 20) 20. The method of any one of items 1 to 19, wherein each dose is between about 10 mg and about 100 mg of the APOC3 RNAi drug substance. (Item 21) 20. The method of any one of items 1 to 19, wherein each dose is between about 10 mg and about 50 mg of the APOC3 RNAi drug substance. (Item 22) 20. The method of any one of items 1 to 19, wherein each dose is between about 25 mg and about 100 mg of the APOC3 RNAi drug substance. (Item 23) 20. The method of any one of items 1 to 19, wherein each dose is between about 25 mg and about 50 mg of the APOC3 RNAi drug substance. (Item 24) 20. The method of any one of items 1 to 19, wherein each dose of the APOC3 RNAi drug substance is about 10 mg. (Item 25) 20. The method of any one of items 1 to 19, wherein each dose of the APOC3 RNAi drug substance is about 25 mg. (Item 26) 20. The method of any one of items 1 to 19, wherein each dose of the APOC3 RNAi drug substance is about 50 mg. (Item 27) 20. The method of any one of items 1 to 19, wherein each dose of the APOC3 RNAi drug substance is about 100 mg. (Item 28) 28. The method of any one of items 1 to 27, wherein the subject is further administered an additional therapeutic agent for the treatment of an APOC3-related disease or disorder. (Item 29) 29. The method of any one of items 1 to 28, wherein the APOC3-related disease or disorder is dyslipidemia. (Item 30) 29. The method of any one of items 1 to 28, wherein the APOC3-related disease or disorder is hypertriglyceridemia, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, lipodystrophy syndrome or familial partial lipodystrophy. (Item 31) 29. The method of any one of items 1 to 28, wherein the APOC3-related disease or disorder is chylomicronemia. (Item 32) 29. The method of any one of items 1 to 28, wherein the APOC3-related disease or disorder is hypertriglyceridemia, with or without a history of pancreatitis. (Item 33) 33. The method according to any one of the preceding claims, wherein the pharmaceutical composition is packaged in a kit, container, pack, dispenser, pre-filled syringe or vial. (Item 34) 34. The method of any one of items 1 to 33, wherein said pharmaceutical composition comprises, consists of, or consists essentially of a formulated APOC3 RNAi drug substance as described in Table 3, consisting of said APOC3 RNAi drug substance in a concentration of about 200 mg / mL, sodium dihydrogen phosphate monohydrate in a concentration of about 0.062 mg / mL, disodium hydrogen phosphate anhydrous in a concentration of about 0.063 mg / mL, and water in a concentration of about 890 mg / mL. (Item 35) 35. The method according to any one of the preceding claims, wherein administration of one or more doses of the pharmaceutical composition is performed by the subject. (Item 36) 36. The method according to any one of the preceding items, wherein the administration of one or more doses of the pharmaceutical composition is performed by a medical professional. (Item 37) 37. The method of any one of items 1 to 36, wherein serum APOC3 protein levels are reduced by more than 60% from baseline levels in the human subject. (Item 38) 38. The method of any one of items 1 to 37, wherein serum triglyceride (TG) levels are reduced by more than 50% from baseline levels in the human subject. (Item 39) 39. The method of claim 38, wherein serum TG levels are reduced by more than 75% from baseline levels in the human subject. (Item 40) 40. The method of any one of items 1 to 39, wherein the serum very low density lipoprotein cholesterol (VLDL-C) level, the serum low density lipoprotein cholesterol (LDL-C) level, or both the serum VLDL-C level and the serum LDL-C level are reduced in the human subject compared to baseline levels. (Item 41) 41. The method of any one of items 1 to 40, wherein the subject has a fasting or postprandial baseline triglyceride level greater than 150 mg / dL. (Item 42) 42. The method of any one of items 1 to 41, wherein the subject has a fasting or postprandial baseline triglyceride level greater than 500 mg / dL. (Item 43) 43. The method of any one of items 1 to 42, wherein the subject has a fasting or postprandial baseline triglyceride level greater than 1000 mg / dL. (Item 44) The method of any one of items 1 to 33 or 35 to 43, wherein the APOC3 RNAi drug substance is administered as a pharma- ceutically acceptable salt, a pharma- ceutically acceptable mixed salt, a free acid, or a combination thereof. (Item 45) 45. The method of claim 44, wherein the APOC3 RNAi drug substance is administered as a pharma- ceutically acceptable sodium salt. (Item 46) A unit dosage form comprising an amount of between about 1 mg and about 100 mg of an APOC3 RNAi drug substance, preferably in a pharma- ceutically acceptable salt form, The RNAi drug substance comprises an antisense strand comprising the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO:2) and a sense strand comprising the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO:6), where a, c, g, i and u represent 2'-O-methyl adenosine, 2'-O-methyl cytidine, 2'-O-methyl guanosine, 2'-O-methyl inosine and 2'-O-methyl uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, 2'-fluoro cytidine, 2'-fluoro guanosine and 2'-fluoro uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue; (NAG37)s represents an inverted abasic deoxyribose residue; [ka] A unit dosage form comprising the structure shown by: (Item 47) 47. The unit dosage form according to item 46, wherein the amount of APOC3 RNAi drug substance is from about 10 mg to about 50 mg. (Item 48) 47. The unit dosage form of item 46, wherein the amount of APOC3 RNAi drug substance is from about 10 mg to about 25 mg. (Item 49) 1. A method of inhibiting expression of APOC3 in a subject in need thereof, comprising administering to the subject an APOC3 RNAi drug substance as set forth in Table 2, comprising administering to the subject one or more doses of the APOC3 RNAi drug substance as set forth in Table 2 in an amount of from about 1 mg to about 100 mg; a. Over the course of about one month; or b. Over the course of about three months; or c. Over the course of approximately six months Administer, method. (Item 50) 50. The method of claim 49, wherein the dose is between about 10 mg and about 100 mg of the APOC3 RNAi drug substance. (Item 51) 50. The method of claim 49, wherein the dose is between about 10 mg and about 50 mg of the APOC3 RNAi drug substance. (Item 52) 50. The method of claim 49, wherein the dose is between about 25 mg and about 100 mg of the APOC3 RNAi drug substance. (Item 53) 50. The method of claim 49, wherein the dose is between about 25 mg and about 50 mg of the APOC3 RNAi drug substance. (Item 54) 50. The method of claim 49, wherein the dose is about 10 mg of the APOC3 RNAi drug substance. (Item 55) 50. The method of claim 49, wherein the dose is about 25 mg of the APOC3 RNAi drug substance. (Item 56) 50. The method of claim 49, wherein the dose is about 50 mg of the APOC3 RNAi drug substance. (Item 57) 50. The method of claim 49, wherein the dose is about 100 mg of the APOC3 RNAi drug substance. (Item 58) A pharmaceutical composition for use in treating an APOC3-related disease or disorder in a human subject in need of such treatment, said use comprising administration of the pharmaceutical composition comprising an APOC3 RNAi agent, wherein the pharmaceutical composition is administered to the subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about one month after the first dose, and wherein the first dose and the second dose are administered by subcutaneous injection. (Item 59) A pharmaceutical composition for use in treating an APOC3-related disease or disorder in a human subject in need of such treatment, wherein the use comprises administration of the pharmaceutical composition comprising an APOC3 RNAi agent, wherein the pharmaceutical composition is administered to the subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and wherein the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 3 months after the first dose, and wherein the first dose and the second dose are administered by subcutaneous injection. (Item 60) A pharmaceutical composition for use in treating an APOC3-related disease or disorder in a human subject in need of such treatment, wherein the use comprises administration of the pharmaceutical composition comprising an APOC3 RNAi agent, wherein the pharmaceutical composition is administered to the subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 4 months after the first dose, wherein the first dose and the second dose are administered by subcutaneous injection. (Item 61) A pharmaceutical composition for use in treating an APOC3-related disease or disorder in a human subject in need of such treatment, wherein the use comprises administration of the pharmaceutical composition comprising an APOC3 RNAi agent, wherein the pharmaceutical composition is administered to the subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent, and wherein the pharmaceutical composition is administered to the subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 6 months after the first dose, and wherein the first dose and the second dose are administered by subcutaneous injection. (Item 62) 1. A pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need thereof, said use comprising administration of said pharmaceutical composition comprising an APOC3 RNAi agent; a. the pharmaceutical composition is administered to the human subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent; b. the pharmaceutical composition is administered to the human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about one month after the first dose; c. The pharmaceutical composition is administered to the human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 3 months to about 6 months after the second dose. (Item 63) 1. A pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need thereof, said use comprising administration of said pharmaceutical composition comprising an APOC3 RNAi agent; a. the pharmaceutical composition is administered to the human subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent; b. the pharmaceutical composition is administered to the human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about one month after the first dose; c. the pharmaceutical composition is administered to the human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 6 months after the second dose. Pharmaceutical compositions. (Item 64) 1. A pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need thereof, said use comprising administration of said pharmaceutical composition comprising an APOC3 RNAi agent; a. the pharmaceutical composition is administered to the human subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent; b. the pharmaceutical composition is administered to the human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 1 month to about 6 months after the first dose; c. The pharmaceutical composition is administered to the human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 3 months to about 6 months after the second dose. (Item 65) 1. A pharmaceutical composition for use in treating an APOC3 associated disease or disorder in a human subject in need thereof, said use comprising administration of said pharmaceutical composition comprising an APOC3 RNAi agent; a. the pharmaceutical composition is administered to the human subject at a first dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent; b. the pharmaceutical composition is administered to the human subject at a second dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 3 months to about 6 months after the first dose; c. the pharmaceutical composition is administered to the human subject at a third dose of between about 1 mg and about 100 mg of the APOC3 RNAi agent about 6 months after the second dose. Pharmaceutical compositions. (Item 66) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose of the APOC3 RNAi agent is between about 10 mg and about 100 mg. (Item 67) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose is between about 10 mg and about 50 mg of the APOC3 RNAi agent. (Item 68) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose is between about 25 mg and about 100 mg of the APOC3 RNAi agent. (Item 69) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose is between about 25 mg and about 50 mg of the APOC3 RNAi agent. (Item 70) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose is about 10 mg of the APOC3 RNAi agent. (Item 71) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose is about 25 mg of the APOC3 RNAi agent. (Item 72) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose is about 50 mg of the APOC3 RNAi agent. (Item 73) 66. The pharmaceutical composition of any one of items 58 to 65, wherein each dose is about 100 mg of the APOC3 RNAi agent. (Item 74) 74. The pharmaceutical composition of any one of items 58 to 73, wherein a further therapeutic agent for the treatment of an APOC3-related disease or disorder is administered to the human subject. (Item 75) 75. The pharmaceutical composition according to any one of items 58 to 74, wherein the APOC3-related disease or disorder is dyslipidemia. (Item 76) 75. The pharmaceutical composition according to any one of items 58 to 74, wherein the APOC3 related disease or disorder is hypertriglyceridemia, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, lipid and / or cholesterol metabolic disorders, atherosclerosis, cardiovascular disease, coronary artery disease, hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes mellitus, familial chylomicronemia syndrome (FCS), chylomicronemia, multifactorial chylomicronemia, lipodystrophy syndrome or familial partial lipodystrophy. (Item 77) 75. The pharmaceutical composition of any one of items 58 to 74, wherein the APOC3 related disease or disorder is familial chylomicronemia syndrome, chylomicronemia or polychylomicronemia. (Item 78) 75. The pharmaceutical composition of any one of items 58 to 74, wherein the APOC3 related disease or disorder is hypertriglyceridemia, either with or without a history of pancreatitis. (Item 79) 79. The pharmaceutical composition according to any one of items 58 to 78, packaged in a kit, container, pack, dispenser, pre-filled syringe or vial. (Item 80) 80. The pharmaceutical composition of any one of items 58 to 79, comprising, consisting of, or consisting essentially of a formulated APOC3 RNAi drug substance as described in Table 3, consisting of APOC3 RNAi drug substance in a concentration of about 200 mg / mL, sodium dihydrogen phosphate monohydrate in a concentration of about 0.062 mg / mL, disodium hydrogen phosphate anhydrous in a concentration of about 0.063 mg / mL, and water in a concentration of about 890 mg / mL. (Item 81) 81. The pharmaceutical composition of any one of items 48 to 80, further administered to the subject with further doses administered about one month apart. (Item 82) 81. The pharmaceutical composition of any one of items 48 to 80, further administered to the subject with further doses administered about 3 months apart. (Item 83) 81. The pharmaceutical composition of any one of items 48 to 80, further administered to the subject with further doses administered about 4 months apart. (Item 84) 81. The pharmaceutical composition of any one of items 48 to 80, wherein the pharmaceutical composition is further administered to the subject in an additional dose after the second dose, wherein the additional doses are administered about 6 months apart. (Item 85) 81. The pharmaceutical composition according to any one of items 48 to 80, wherein the pharmaceutical composition is further administered to the subject in an additional dose after the third dose, wherein the additional doses are administered about 3 to about 6 months apart. (Item 86) 86. The pharmaceutical composition according to any one of items 48 to 85, wherein administration of one or more doses of the pharmaceutical composition is performed by the subject. (Item 87) 86. The pharmaceutical composition according to any one of items 48 to 85, wherein administration of the one or more doses of the pharmaceutical composition is performed by a medical professional. (Item 88) 88. The pharmaceutical composition according to any one of items 48 to 87, wherein serum APOC3 protein levels are reduced by more than 60% in the human subject compared to baseline levels. (Item 89) 89. The pharmaceutical composition of any one of items 48 to 88, wherein serum triglyceride (TG) levels are reduced by more than 50% compared to baseline levels in the human subject. (Item 90) 90. The pharmaceutical composition of item 89, wherein serum TG levels are reduced by more than 75% compared to baseline levels in the human subject. (Item 91) 91. The pharmaceutical composition of any one of items 48 to 90, wherein the serum very low density lipoprotein cholesterol (VLDL-C) level, the serum low density lipoprotein cholesterol (LDL-C) level, or both the serum VLDL-C level and the serum LDL-C level are reduced in the human subject compared to baseline levels. (Item 92) 92. The pharmaceutical composition of any one of paragraphs 48-79 or 81-91, wherein the APOC3 RNAi agent is administered as a salt, a mixed salt, a free acid, or a combination thereof. (Item 93) 93. The pharmaceutical composition of claim 92, wherein the APOC3 RNAi agent is administered as a pharma- ceutical acceptable sodium salt. (Item 94) 1. A pharmaceutical composition for inhibiting expression of APOC3 in a human subject in need thereof by administration of a pharmaceutical composition comprising an APOC3 RNAi agent, comprising one or more doses of the APOC3 RNAi agent in an amount of about 1 mg to about 100 mg, a. Over the course of about one month, or b. Over the course of about three months; or c. Over the course of approximately six months The pharmaceutical composition is administered. (Item 95) 95. The pharmaceutical composition of item 94, wherein the dose is between about 10 mg and about 100 mg of the APOC3 RNAi agent. (Item 96) 95. The pharmaceutical composition of item 94, wherein the dose is between about 10 mg and about 50 mg of the APOC3 RNAi agent. (Item 97) 95. The pharmaceutical composition of item 94, wherein the dose is between about 25 mg and about 100 mg of the APOC3 RNAi agent. (Item 98) 95. The pharmaceutical composition of item 94, wherein the dose is between about 25 mg and about 50 mg of the APOC3 RNAi agent. (Item 99) 95. The pharmaceutical composition of claim 94, wherein the dose is about 10 mg of the APOC3 RNAi agent. (Item 100) 95. The pharmaceutical composition of claim 94, wherein the dose is about 25 mg of the APOC3 RNAi agent. (Item 101) 95. The pharmaceutical composition of claim 94, wherein the dose is about 50 mg of the APOC3 RNAi agent. (Item 102) 95. The pharmaceutical composition of claim 94, wherein the dose is about 100 mg of the APOC3 RNAi agent.

Claims

[Claim 1] The invention described in the specification.

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