Methods for the Management of APOC3-Related Diseases and Disorders

Abstract

The method described herein comprises the pharmaceutical composition of APOC3 RNAi agent for treating the disease and disorder that can be partially mediated by the reduction of APOC3 gene expression in the human subject that needs treatment.When the pharmaceutical composition is administered to the human subject that needs treatment according to the method disclosed herein, it treats the disease and disorder that is related to elevated triglyceride (TG) level, for example, lipodystrophy syndrome, including familial chylomicronemia syndrome (FCS), hypertriglyceridemia, obesity, dyslipidemia, non-alcoholic fatty liver disease, non-alcoholic fatty liver disease, hyperlipidemia, lipid and / or cholesterol metabolic disorder, atherosclerosis, cardiovascular disease, coronary artery disease, hypertriglyceridemia-induced pancreatitis, metabolic syndrome, type II diabetes, chylomicronemia, multifactorial chylomicronemia or familial partial lipodystrophy.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 883,046, filed Aug. 5, 2019; U.S. Provisional Patent Application No. 62 / 936,559, filed Nov. 17, 2019; and U.S. Provisional Patent Application No. 62 / 970,613, filed Feb. 5, 2020, the contents of each of which are hereby incorporated by reference in their entirety.

[0002] Sequence Listing This application includes a sequence listing submitted in ASCII format, which is hereby incorporated by reference in its entirety. This ASCII copy has a name of 30683_WO_SequenceListing.txt and a size of 3 kb.

[0003] Field of the Invention Disclosed herein are methods for the treatment of diseases and disorders that can be mediated, at least in part, by silencing or inhibiting apolipoprotein C - III (also referred to as APOC3, apoC - III, APOC - III, and APO C - III) gene expression using a pharmaceutical composition comprising an RNA interference (RNAi) agent that inhibits APOC3 gene expression.

Background Art

[0004] Background Apolipoprotein C-III (also known as 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 a contributing factor in the development of atherosclerosis. Individuals with severe hypertriglyceridemia (often >1000 mg / dL) are also at risk of 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 consist of a hydrophobic triacylglycerol and cholesteryl ester core, as well as a hydrophilic outer layer of phospholipids, cholesterol, and apolipoproteins. APOC3 is one of these apolipoproteins.

[0005] APOC3 is primarily synthesized in the liver and plays a crucial role in the production, metabolism, and clearance of TG-rich lipoproteins from plasma. Several gain-of-function polymorphisms (GMOs) that are thought to contribute to the development of hypertriglyceridemia have been identified in the promoter region of the APOC3 gene (see, for example, 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. Furthermore, the excess of APOC3 further increases serum TG levels by inhibiting the activity of lipoprotein lipases and hepatic lipases, and by delaying the catabolism of TG-rich lipoproteins. In addition, elevated APOC3 also delays 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 in 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 oil in moderate cases, but in most cases, the reduction in serum TG is modest. Furthermore, available therapeutics are often ineffective in patients with a monogenic cause 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-tethered high-density lipoprotein-binding protein 1 (GPIHBP1) are less frequently observed. These mutations result in dysfunctional lipoprotein lipase, but functional lipoprotein lipase is required for an optimal response to standard treatment. There is a need for effective therapeutic agents that can provide substantial triglyceride-reducing effects for the treatment of conditions 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, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, hypertriglyceridemia, lipid and / or cholesterol metabolism disorders, atherosclerosis, cardiovascular disease, coronary artery disease, and other dyslipidemia and metabolism-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) [Overview of the project] [Means for solving the problem]

[0008] overview A method for treating APOC3-related diseases and disorders in human subjects requiring such treatment is described herein, comprising the step of administering to a human subject a pharmaceutical composition containing the compositions listed in Table 2 (i.e., APOC3 RNAi drug substances, also referred to herein as ADS-005) in doses between approximately 1 mg and approximately 100 mg of the APOC3 RNAi drug substance, wherein the pharmaceutical composition is administered subcutaneously and there is approximately one month between doses (i.e., monthly dosing). In some embodiments, the pharmaceutical composition used in the method disclosed herein contains, consists of, or essentially consists of, the formulated APOC3 RNAi drug substance listed in Table 3 (also referred to herein as ADS-005-1).

[0009] The APOC3 RNAi drug substances described herein provided unexpected potency and duration of effect. As shown in the data from human clinical studies presented herein, embodiments of APOC3 RNAi drug substances at single doses of less than 100 mg provided sustained inhibition of APOC3 gene expression that lasted for several months, resulting in substantial reductions in triglycerides (TG) and other relevant lipid parameters, while maintaining a favorable safety profile. This has led to the discovery of novel low-dose and low-frequency dosing regimens for the treatment of APOC3-related diseases and disorders.

[0010] The treatment methods described herein result in a reduction in APOC3 expression in human subjects, which in turn leads to a reduction in serum triglyceride (TG) levels in the subjects.

[0011] Furthermore, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment for such a disease or disorder is described herein, comprising the step of administering to the human subject a pharmaceutical composition containing an APOC3 RNAi drug substance (i.e., ADS-005) listed in Table 2 in doses between approximately 1 mg and approximately 50 mg, wherein the pharmaceutical composition is administered subcutaneously and there is approximately one month between dose administrations (i.e., monthly dosing).

[0012] A method for treating APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is further described herein, comprising the step of administering to the human subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in a dose between approximately 1 mg and approximately 100 mg, wherein the pharmaceutical composition is administered subcutaneously and there is approximately 3 months between dose administrations (e.g., 4 doses per year or 12-week doses (q12w)).

[0013] A method for treating APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is also described herein, comprising the step of administering to the human subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in a dose between approximately 1 mg and approximately 50 mg, wherein the pharmaceutical composition is administered subcutaneously and there is approximately 3 months between dose administrations (e.g., 4 doses per year or 12-week doses (q12w)).

[0014] A method for treating APOC3-related disease or disorder in a human subject requiring treatment for such disease or disorder is described herein, comprising the step of administering to the human subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in doses between approximately 1 mg and approximately 100 mg, wherein the pharmaceutical composition is administered subcutaneously, and a second dose is administered approximately one month after the initial dose, with approximately three months between subsequent doses.

[0015] A method for treating APOC3-related disease or disorder in a human subject requiring treatment for such disease or disorder is described herein, comprising the step of administering to the human subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in a dose between approximately 1 mg and approximately 50 mg, wherein the pharmaceutical composition is administered subcutaneously, and the initial dose is followed by a second dose approximately one month later, with a dose interval of approximately three months between subsequent doses.

[0016] A method for treating an APOC3-related disease or disorder in a human subject who needs treatment for the APOC3-related disease or disorder, the method comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance at a dose between about 1 mg and about 100 mg as set forth in Table 2, the pharmaceutical composition being administered subcutaneously, and there being about 4 months between dose administrations, the method being further described herein.

[0017] A method for treating an APOC3-related disease or disorder in a human subject who needs treatment for the APOC3-related disease or disorder, the method comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance at a dose between about 1 mg and about 50 mg as set forth in Table 2, the pharmaceutical composition being administered subcutaneously, and there being about 4 months between dose administrations, the method also being described herein.

[0018] A method for treating an APOC3-related disease or disorder in a human subject who needs treatment for the APOC3-related disease or disorder, the method comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance at a dose between about 1 mg and about 100 mg as set forth in Table 2, the pharmaceutical composition being administered subcutaneously, and after the first dose, a second dose follows about 1 month later, and thereafter, for subsequent doses, there is about 4 months between dose administrations, the method being described herein.

[0019] A method for treating an APOC3-related disease or disorder in a human subject who needs treatment for the APOC3-related disease or disorder, the method comprising administering to the human subject a pharmaceutical composition comprising an APOC3 RNAi drug substance at a dose between about 1 mg and about 50 mg as set forth in Table 2, the pharmaceutical composition being administered subcutaneously, and after the first dose, a second dose follows about 1 month later, and thereafter, for subsequent doses, there is about 4 months between dose administrations, the method being described herein.

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

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

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

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

Brief Description of the Drawings

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

[0025] [Figure 2] Chemical structures of APOC3 RNAi drug substances listed in Table 2, shown in sodium salt form.

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

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

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

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

[0030] [Figure 7] Graph showing serum APOC3 protein levels in healthy human volunteers who received either placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug (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 who received either 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 who received either 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 who received either 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] A graph showing serum triglyceride (TG) levels in healthy human volunteers who received either placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug (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 who received either 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 who received either 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 who received either placebo (all cohorts) or a single 25 mg dose of APOC3 RNAi drug (cohort 1) from the Phase I clinical study described in Example 2.

[0038] [Figure 15]A graph showing serum VLDL-C levels in healthy human volunteers who received either placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug (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 who received either placebo (all cohorts) or a single 100 mg dose of APOC3 RNAi drug (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 who received either 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 ultra-high density lipoprotein cholesterol (HDL-C) levels in healthy human volunteers who received either 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 who received either placebo (all cohorts) or a single 50 mg dose of APOC3 RNAi drug (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 who received either 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 who received either placebo (all cohorts) or a single 10 mg dose of APOC3 RNAi drug (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 who received either placebo (all cohorts) or a single 25 mg dose of APOC3 RNAi drug (cohort 1) from the Phase I clinical study described in Example 2.

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

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

[0048] [Figure 25] Graph showing serum LDL-C levels in healthy human volunteers who received either 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]A graph showing serum APOC3 protein levels in patients with chylomicronemia who received a 50 mg dose of APOC3 RNAi drug (Cohort 5) from the Phase I clinical study described in Example 2.

[0050] [Figure 27] A graph showing serum triglyceride levels in patients with chylomicronemia who received a 50 mg dose of APOC3 RNAi drug (Cohort 5) from the Phase I clinical study described in Example 2. [Modes for carrying out the invention]

[0051] Detailed explanation RNAi agents The methods described herein include the administration of a pharmaceutical composition to a human subject, the pharmaceutical composition comprising 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 the administration of a pharmaceutical composition to a human subject, the pharmaceutical composition comprising an APOC3 RNAi drug substance (also referred to as ADS-005) as listed in Table 2. A suitable composition for use in the methods disclosed herein comprises 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 comprises a nucleotide sequence provided in Tables 1A and 1B, the sense strand of the RNAi agent is further ligated or conjugated to a targeting group comprising three N-acetyl-galactosamine targeting moieties (see, for example, Table B). The RNAi agent that inhibits the expression of the APOC3 gene in a human subject is referred to as the "APOC3 RNAi agent".

[0052] Generally, APOC3 RNAi agents include a sense strand (also called a passenger strand) and an antisense strand (also called a guide strand) that anneal to form a double helix. The APOC3 RNAi agents disclosed herein include RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecules capable of degrading or inhibiting the translation of messenger RNA (mRNA) transcripts of APOC3 mRNA in a sequence-specific manner. The APOC3 RNAi agents disclosed herein may act via RNA interference mechanisms (i.e., inducing RNA interference through interaction with the RNA interference pathway machine (RNA-induced silencing complex or RISC) in mammalian cells) or by any alternative mechanism or pathway. While APOC3 RNAi agents are considered to act primarily via RNA interference mechanisms as used herein, the disclosed RNAi agents are not constrained by or limited to any particular pathway or mechanism of action. RNAi agents generally consist of a sense strand and an antisense strand, each 16 to 49 nucleotides long, 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 sense strand of an APOC3 RNAi agent is typically 16–49 nucleotides long, and the antisense strand is typically 18–49 nucleotides long. In some embodiments, the sense and antisense strands are independently 17–26 nucleotides long. In some embodiments, the sense and antisense strands are independently 21–26 nucleotides long. In some embodiments, the sense and antisense strands are independently 21–24 nucleotides long. In some embodiments, the sense and / or antisense strands are independently 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long. In some embodiments, both the sense and antisense strands are 21 nucleotides long. The sense and antisense strands may be the same length or different lengths. The sense and antisense strands may also form overhanging nucleotides at one or both ends of the APOC3 RNAi agent.

[0054] APOC3 RNAi agents inhibit, silence, or knock down APOC3 gene expression. As used herein, the terms “silencing,” “reducing,” “inhibiting,” “downregulating,” or “knockdown” mean, when referring to APOC3 expression, that the expression of the gene, as measured by the level of RNA transcribed from the gene or the level of polypeptides, proteins, or protein subunits translated from mRNANA, is reduced when the cells, cell populations, tissues, organs, or subjects are treated with the RNAi agent compared to a second set of cells, cell populations, tissues, organs, or subjects that have not been treated in the same way. In some cases, 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 containing the APOC3 RNAi agent with the APOC3 mRNA or APOC3 protein level after administration of the therapeutic agent.

[0055] Inhibition, silencing, or knockdown of the 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), which is incorporated herein by reference in its entirety, provide some specific examples of suitable assays for measuring APOC3 gene expression inhibition. A reference APOC3 mRNA gene transcript for a normal human (referred to as transcript variant 1; GenBank NM_000040.1) can be found in Sequence 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 containing 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 containing one or more N-acetyl-galactosamine moieties, thereby forming the APOC3 RNAi drug substances listed in Table 2. In some embodiments, the methods described herein include the administration of the APOC3 RNAi drug substances listed in Table 2. The APOC3 RNAi drug substances listed in Table 2 include the APOC3 RNAi agents shown in Table 1A (antisense strand) and Table 1B (sense strand). The N-acetyl-galactosamine moieties facilitate the targeting of the APOC3 RNAi agent to asialoglycoprotein receptors (ASGPr) readily present on the surface of hepatocytes, resulting in the 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 an antisense strand having a region complementary to at least a portion of APOC3 mRNA. APOC3 RNAi agents and APOC3 RNAi drug substances suitable for use in the disclosed methods are described in their entirety in the previously mentioned International Patent Application Publication No. WO2019 / 051402 (Patent Application No. PCT / US2018 / 050248), which is thus incorporated herein by reference.

[0058] As used herein, the terms “sequence” and “nucleotide sequence” mean a sequence or order of nucleic acid bases or nucleotides, described using standard terminology and a sequence of letters. As used herein, the terms “nucleic acid base” and “nucleotide” have the same meanings as commonly understood in the art.

[0059] As used herein, the term “complementary” means, 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 the targeted mRNA sequence), the ability of an oligonucleotide containing a first nucleotide sequence to hybridize (form base-pair hydrogen bonds) under mammalian physiological conditions (or other suitable conditions) and to form a double-stranded or double-helical structure with an oligonucleotide containing a second nucleotide sequence under certain standard conditions. Those skilled in the art can select the set of conditions most appropriate for hybridization testing. The complementary sequence includes Watson-Crick base pairs or non-Watson-Crick base pairs and includes, at a minimum, natural or modified nucleotides or nucleotide mimeographs 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 the purpose of determining identity or complementarity.

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

[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 the contiguous sequence of the first oligonucleotide hybridize with the same number of bases in the contiguous sequence of the second polynucleotide.

[0062] As used herein, “substantially complementary” means that in a hybridized pair of nucleotide sequences, at least 85% of, 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 in reference to nucleotide matching between the sense and antisense strands of an RNAi agent, or between the antisense strand of an RNAi agent and the sequence of APOC3 mRNA.

[0063] As used herein, the terms “substantially identical” or “substantially identical,” when applied to nucleic acid sequences, mean that a given nucleic acid sequence contains a sequence with at least about 85% or higher sequence identity compared to a reference sequence, e.g., at least 90%, at least 95%, or at least 99%. The percentage of sequence identity is determined by comparing two optimally aligned sequences across a comparison window. The percentage is calculated by determining the number of positions in which identical nucleic acid bases exist in both sequences to obtain the number of matching positions, dividing the number of matching 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 inventions disclosed herein encompass nucleotide sequences that are substantially identical to those disclosed herein.

[0064] Linking between modified nucleotides and modified nucleosides The APOC3 RNAi agents disclosed herein may consist of modified nucleotides that can maintain RNAi agent activity while simultaneously increasing serum stability, and that can minimize the possibility of activating interferon activity in humans. As used herein, “modified nucleotide” refers to a nucleotide other than a ribonucleotide (2'-hydroxylnucleotide). In some embodiments, at least 50% (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%) of the nucleotides are modified nucleotides. As used herein, modified nucleotides include, but are not limited to, any known modified nucleotides known in the art, including, deoxyribonucleotides, nucleotide mimes, 2'-modified nucleotides, inverted nucleotides, modified nucleic acid base-containing nucleotides, cross-linked nucleotides, peptide nucleic acids (PNAs), 2',3'-seconucleotide mimes (unlocked nucleic acid base analogs), locked nucleotides, 3'-O-methoxy(2'-nucleoside-linked) nucleotides, 2'-F-arabinonucleotides, 5'-Me, 2'-fluoronucleotides, morpholinonucleotides, vinylphosphonate-containing nucleotides, and cyclopropylphosphonate-containing nucleotides. In some embodiments, the modified nucleotide of the APOC3 RNAi agent is a 2'-modified nucleotide (i.e., a nucleotide having a group other than a hydroxyl group at the 2' position of a five-membered sugar ring). 2'-modified nucleotides include, but are not limited to, 2'-O-methylnucleotides, 2'-deoxy-2'-fluoronucleotides (commonly referred to simply as 2'-fluoronucleotides), 2'-deoxynucleotides, 2'-methoxyethyl (2'-O-2-methoxyethyl) nucleotides, 2'-aminonucleotides, and 2'-alkylnucleotides. Further 2'-modified nucleotides are known in the art. Not all nucleotides in a given RNAi agent need to be uniformly modified. Furthermore, more than one modification may be incorporated into a single APOC3 RNAi agent, or even into its single nucleotide.The sense and antisense strands of APOC3 RNAi agents can be synthesized and / or modified by methods known in the art. Modifications at one nucleotide are independent of modifications at other nucleotides.

[0065] In some embodiments, nucleic acid bases (often simply referred to as "bases") can be modified. As commonly used in the art, natural nucleic acid bases include the major purine bases adenine and guanine, and the major pyrimidine bases cytosine, thymine, and uracil. Nucleic acid bases 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 nucleic acid bases (including phosphoramidite compounds containing modified nucleic acid bases) is known in the art.

[0066] Modified nucleic acid bases 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- This includes thiocytosine, 5-halouracil, cytosine, 5-propynyluracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothimine, 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 the 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 having four or fewer (i.e., 0, 1, 2, 3, or 4) nucleotides that are ribonucleotides (i.e., unmodified) in both the sense strand and the antisense strand. As used herein, a sense strand in which substantially all of the nucleotides present are modified nucleotides is a sense strand having two 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 having two 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 the APOC3 RNAi agent are linked by non-standard linkages or skeletons (i.e., modified nucleoside linkages or modified skeletons). Modified nucleoside linkages or skeletons include, but are not limited to, phosphorothioate groups, chiral phosphorothioates, thiophosphates, phosphorodithioates, phosphotriesters, aminoalkyl-phosphotriesters, alkylphosphonates (e.g., methylphosphonate or 3'-alkylenephosphonate), chiral phosphonates, phosphinates, phosphoramidates (e.g., 3'-aminophosphoramide, aminoalkylphosphoramide, or thionophosphoramide), thionoalkyl-phosphonates, thionoalkylphosphotriesters, morpholino linkages, boranophosphates having the usual 3'-5' linkage, 2'-5' linked analogs of boranophosphates, or boranophosphates having inverted polarity where adjacent pairs of nucleoside units are linked from 3'-5' to 5'-3' or 2'-5' to 5'-2'. In some embodiments, the modified internucleoside linkages or skeletons lack a phosphorus atom. Modified internucleoside linkages lacking a phosphorus atom include, but are not limited to, short-chain alkyl or cycloalkyl sugar linkages, mixed heteroatoms and alkyl or cycloalkyl sugar linkages, or one or more short-chain heteroatoms or heterocyclic sugar linkages. In some embodiments, the modified internucleoside skeletons include, but are not limited to, siloxane skeletons, sulfide skeletons, sulfoxide skeletons, sulfone skeletons, formacetyl and thioformacetyl skeletons, methyleneformacetyl and thioformacetyl skeletons, alkene-containing skeletons, sulfamate skeletons, methyleneimino and methylenehydrazino skeletons, sulfonate and sulfonamide skeletons, amide skeletons, and other skeletons having mixed N, O, S and CH2 components.

[0069] In some embodiments, the sense strand of the APOC3 RNAi agent may contain 1, 2, 3, 4, 5, or 6 phosphorothioate linkages, the antisense strand of the 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 the APOC3 RNAi agent may contain 1, 2, 3, or 4 phosphorothioate linkages, the antisense strand of the 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 nucleoside linkages. In some embodiments, the at least two phosphorothioate nucleoside linkages are located between nucleotides 1-3 from the 3' end of the sense strand. In some embodiments, the at least two phosphorothioate nucleoside linkages are located 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, the phosphorothioate nucleoside linkages are used to link terminal nucleotides in the sense strand to capping residues present at the 5' end, 3' end, or both the 5' and 3' ends of the nucleotide sequence. In some embodiments, the phosphorothioate nucleoside linkages are used to link targeting groups to the sense strand.

[0071] In some embodiments, the antisense strand of the APOC3 RNAi agent contains three or four phosphorothioate nucleoside linkages. In some embodiments, the antisense strand of the APOC3 RNAi agent contains three phosphorothioate nucleoside linkages. In some embodiments, the three phosphorothioate nucleoside linkages are located 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. In some embodiments, the APOC3 RNAi agent contains at least two phosphorothioate nucleoside linkages in the sense strand and three or four phosphorothioate nucleoside linkages in the antisense strand.

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

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

[0074] In some embodiments, one or more inverted abasid residues (invAbs) are added to the 3' end of the sense strand. In some embodiments, one or more inverted abasid residues (invAbs) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasid residues or sites are inserted between the nucleotide sequence of the targeted ligand and the sense strand of the RNAi agent. In some embodiments, including one or more inverted abasid residues or sites at or near the end(s) of the sense strand of the RNAi agent enables enhanced activity or other desired properties of the RNAi agent.

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

[0076] [Table A]

[0077] Targeted portion and base 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 can enhance the targeting or delivery of the RNAi agent. 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 listed in Table 2 herein, including three N-acetyl-galactosamine targeting moieties, are provided in Table B. Targeting moieties or targeting groups may be covalently ligated to the 3' and / or 5' ends of either the sense strand and / or antisense strand. In some embodiments, the APOC3 RNAi agent contains a targeting group ligated to the 3' and / or 5' ends of the sense strand. In some embodiments, the targeting group is ligated to the 5' end of the sense strand of the APOC3 RNAi agent. In some embodiments, the targeting group comprises, essentially consists of, or is composed of, the structure (NAG37)s and is ligated to the 5' end of the sense strand of the APOC3 RNAi agent. The targeting group may be ligated to the RNAi agent directly or indirectly via a linker / binding group. In some embodiments, the targeting group is ligated to the RNAi agent via an unstable, cleavable, or reversible bond or linker. In some embodiments, the targeting group is ligated to an inverted debase residue at the 5' end of the sense strand.

[0078] Targeting groups or moieties can enhance the pharmacokinetic or in vivo distribution characteristics of the conjugate or RNAi agent to which they are bound, thereby improving the 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, divalent, trivalent, tetravalent, or have a higher valency depending on the target they are directed to. Typical 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 asialoclycoprotein receptor ligand. In some embodiments, the asialoclycoprotein receptor ligand comprises or consists of one or more galactose derivatives. As used herein, the term galactose derivative includes both galactose and derivatives of galactose having an affinity for the asialoclycoprotein receptor equal to or higher than that of galactose. Galactose derivatives include, but are not limited to, galactose, galactosamine, N-formylgalactosamine, N-acetylgalactosamine, N-propionylgalactosamine, Nn-butanoylgalactosamine, and N-isobutanoylgalactosamine (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 into the liver are well known in the field (see, for example, 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 via their binding to asialoglycoprotein receptors expressed on the surface of hepatocytes. Binding of asialoglycoprotein receptor ligands to asialoglycoprotein receptors(s) facilitates cell-specific targeting to hepatocytes and endocytosis of molecules into hepatocytes. Asialoglycoprotein receptor ligands can be monomeric (e.g., having a single galactose derivative) or multimeric (e.g., having multiple galactose derivatives). Galactose derivatives or galactose derivative "clusters" can be bound 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 2 to 4 terminal galactose derivatives. The terminal galactose derivatives are bonded to the molecule via their C-1 carbon. In some embodiments, the galactose derivative cluster is a galactose derivative trimer (also called a tri-antennary galactose derivative or trivalent galactose derivative). In some embodiments, the galactose derivative cluster comprises 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 called a tetra-antennary galactose derivative or 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. The galactose derivatives may be linked to the central branch point via the C-1 carbon of a saccharide. In some embodiments, the galactose derivatives are 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, e.g., U.S. Patent No. 5,885,968; Biessen et al. J. Med. Chem. 1995 Vol. 39 p. 1538-1546). The branch point can be any small molecule that enables the linkage of the three galactose derivatives and further enables the linkage of the branch point to an RNAi agent. Examples of branch point groups are di-lysine or di-glutamate. The binding of the branching point to the RNAi agent may occur via a linker or spacer. In some embodiments, the linker or spacer includes, but is not limited to, a flexible hydrophilic spacer, such as a PEG spacer. In some embodiments, the linker includes a rigid linker, such as a cyclic group. In some embodiments, the galactose derivative includes or consists of N-acetyl-galactosamine. In some embodiments, the galactose derivative cluster consists of a galactose derivative tetramer, which may be, for example, an N-acetyl-galactosamine tetramer.

[0083] Preparations of galactose derivative clusters containing a targeting group, such as N-acetyl-galactosamine, are 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 thus incorporated herein by reference in their entirety.

[0084] For example, the targeted ligands conjugated to the APOC3 RNAi agents listed in Tables 1A and 1B have the chemical structure of (NAG37)s shown in Table B below.

[0085] [Table B]

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

[0087] [Table 1A]

[0088] [Table 1B]

[0089] When used in Tables 1A, 1B, and 2 of this specification, the following notations are used to indicate modified nucleotides, targeting groups, and linking groups: A, C, G, I, and U represent adenosine, cytidine, guanosine, inosine, and uridine, respectively; a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasal deoxyribose residue (see Table A); (NAG37)s represents 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 the phosphorothioate linkage "s"), monomers are linked to each other by a 5'-3'-phosphodiester bond when present in a chain. Those skilled in the art will clearly understand that including a phosphorothioate linkage, as shown, in the modified nucleotide sequences disclosed herein replaces the phosphodiester linkage typically present in oligonucleotides (see, for example, Figures 1-3 showing all linkages). Furthermore, those skilled in the art will readily understand that the terminal nucleotide at the 3' end of a given oligonucleotide sequence typically has a hydroxyl (-OH) group at the 3' position of each given monomer in ex vivo, instead of a phosphate moiety. Furthermore, in the embodiments disclosed herein, when each chain is viewed 5'→3', the inverted debasic residue is inserted such that the 3' position of deoxyribose is linked at the 3' end of the preceding monomer on each chain. Furthermore, as will be readily understood and recognized by those skilled in the art, the phosphorothioate chemical structures shown herein typically represent an anion on a sulfur atom, but the inventions disclosed herein encompass all phosphorothioate tautomers (for example, when the sulfur atom has a double bond and the anion is an oxygen atom). Unless otherwise expressly indicated herein, such understanding by those skilled in the art is used when describing the APOC3 RNAi agents and compositions comprising APOC3 RNAi agents disclosed herein.

[0091] Each sense strand and / or antisense strand may have any of the above-listed targeting or linking groups, as well as other targeting or linking groups, conjugated at 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 human subjects, thereby silencing the translation of the APOC3 protein using an RNA interference mechanism for human subjects possessing APOC3.

[0093] In some embodiments, the methods disclosed herein use APOC3 RNAi drug substances 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 the complete chemical structure is shown in Figures 1A-1D (free acid form) and Figures 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 a sodium salt.

[0097] Pharmaceutical compositions and formulations APOC3 RNAi agents suitable for use in the methods disclosed herein may be prepared as pharmaceutical compositions or formulations for administration to human subjects. Pharmaceutical compositions may be used to treat subjects with diseases or disorders that would benefit from inhibition of APOC3 mRNA expression or reduction in APOC3 protein levels, e.g., human subjects with APOC3-related diseases or disorders. In some embodiments, the method includes the step of administering an APOC3 RNAi agent linked to a targeting group or targeting ligand described herein to a subject requiring treatment. In some embodiments, one or more pharmaceutically acceptable excipients (including vehicles, carriers, diluents, and / or delivery polymers) are added to the pharmaceutical composition containing the APOC3 RNAi agent to form a pharmaceutical formulation suitable for in vivo delivery to human subjects.

[0098] When administered to a human subject using the method disclosed herein, a pharmaceutical composition containing an APOC3 RNAi agent reduces or diminishes 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 the clinical presentation in subjects with APOC3-related disease or disorder. In some embodiments, one or more pharmaceutical compositions in a therapeutic or prophylactic dose are administered to subjects requiring such treatment. In some embodiments, administration of any of the disclosed APOC3 RNAi agents may be used to reduce the number, severity, and / or frequency of disease symptoms in a subject.

[0100] A pharmaceutical composition comprising an APOC3 RNAi agent may be used to treat at least one symptom in a subject having a disease or disorder that benefits from reduction or inhibition of APOC3 protein expression levels. In some embodiments, the subject is administered one or more pharmaceutical compositions comprising an APOC3 RNAi agent in a therapeutically effective dose to treat the symptom. In other embodiments, the subject is administered one or more APOC3 RNAi agents in a prophylactically effective dose to prevent at least one symptom.

[0101] The APOC3 RNAi agents disclosed herein may be administered via any suitable route in preparations appropriately modified for a specific route. Therefore, the pharmaceutical compositions described herein may be administered by injection, for example, intravenously or subcutaneously. In some embodiments, the pharmaceutical compositions described herein are administered by subcutaneous injection.

[0102] When used herein, a pharmaceutical composition or pharmaceutically acceptable excipient comprises a pharmacologically effective amount of at least one APOC3 RNAi agent and one or more pharmaceutically acceptable excipients. A pharmaceutically acceptable excipient 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. Excipients do not exert a therapeutic effect at the intended dosage, nor are they intended to exert a therapeutic effect. Excipients may act to a) assist in the processing of the drug delivery system during manufacturing, b) protect, support, or enhance the stability, bioavailability, or patient acceptability of the API, c) assist in the identification of the product, and / or d) enhance any other properties of the overall safety, efficacy, or delivery of the API during storage or use. A pharmaceutically acceptable excipient may be an inert substance or not.

[0103] Excipients may include, but are not limited to, absorption enhancers, anti-adhesives, defoamers, antioxidants, binders, buffers, carriers, coatings, pigments, delivery enhancers, delivery polymers, dextran, dextrose, diluents, disintegrants, emulsifiers, bulking agents, fillers, flavoring and deodorizing 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® EL™ (BASF, Parsippany, NJ), or phosphate-buffered saline (PBS). These must be stable under manufacturing and storage conditions and should be protected from contamination by microorganisms such as bacteria and fungi. Carriers may be solvents or dispersion media containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof.

[0105] Injectable sterile solutions can be prepared by incorporating the active compound in the required amount into a suitable solvent along with one or a combination thereof of the components listed above, and then sterilizing by filtration as necessary. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and any other required components from those listed above.

[0106] In some embodiments, a pharmaceutical composition suitable for use in the methods disclosed herein comprises components identified in the formulated APOC3 RNAi drug substance provided in Table 3 below.

[0107] APOC3 RNAi agents can be formulated in a dose unit form within a composition for ease of administration and uniformity of dosage. A dose unit form refers to a physically distinct unit suitable as a unit dose for the subject being treated; each unit contains a predetermined amount of the active compound calculated to produce the desired therapeutic effect in relation to the required pharmaceutical carrier. In some embodiments, the dose unit is between about 1 mg and about 100 mg of APOC3 RNAi drug substance. In some embodiments, the dose unit is between about 10 mg and about 100 mg of APOC3 RNAi drug substance. In some embodiments, the dose unit is between about 10 mg and about 50 mg of APOC3 RNAi drug substance. In some embodiments, the dose unit is about 10 mg of APOC3 RNAi drug substance. In some embodiments, the dose unit is about 25 mg of APOC3 RNAi drug substance. In some embodiments, the dose unit is about 50 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is approximately 100 mg of APOC3 RNAi drug substance. In some embodiments, the dosage unit is approximately 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. This refers to APOC3 RNAi drug substances in amounts of approximately 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.

[0108] The pharmaceutical composition may include other further components commonly found in pharmaceutical compositions. Such further components include, but are not limited to, antipruritic agents, astringents, topical anesthetics, or anti-inflammatory agents (e.g., antihistamines, diphenhydramine, etc.).

[0109] As used herein, “pharmacologically effective dose,” “therapeutic effective dose,” or simply “effective dose” refers to the amount of an RNAi agent that produces a pharmacological, therapeutic, or prophylactic effect.

[0110] The pharmaceutically acceptable formulations described herein may be packaged in kits, containers, packs, or dispensers. The pharmaceutical compositions described herein may be packaged in pre-filled syringes or vials.

[0111] Formulated APOC3 RNAi drug substance In some embodiments, the APOC3 RNAi drug substance (ADS-005) provided in Table 2 is formulated with one or more pharmaceutically acceptable excipients to form a pharmaceutical composition suitable for administration to a human subject. In some embodiments, the APOC3 RNAi drug substance listed 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 formulated APOC3 RNAi drug substances according to Table 3 are prepared as sterile formulations. In some embodiments, the formulated APOC3 RNAi drug substances are packaged in a container, for example, a glass vial. In some embodiments, the formulated APOC3 RNAi drug substances are packaged in a glass vial having a filling volume of about 1.2 mL, and the desired volume for administration may be calculated based on the desired dose level to be administered.

[0114] In some embodiments, the formulated APOC3 RNAi drug substances shown in Table 3 are administered to human subjects using the methods disclosed herein.

[0115] Human subjects with elevated triglyceride levels and / or APOC3 overexpression The methods disclosed herein include treating a disease, disorder, or other condition or symptom in a human subject requiring the use of a pharmaceutical composition containing an APOC3 RNAi drug substance as listed in Table 2, which can be at least partially ameliorated by reduction or silencing of APOC3 gene expression. In some embodiments, the methods disclosed herein include treating a disease or condition associated with elevated triglycerides in a human subject requiring the use of a pharmaceutical composition containing an APOC3 RNAi drug substance as listed 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, human subjects are 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 rare, severe genetic disorder with a prevalence of 1 in 1,000,000, often caused by various monogenic mutations (e.g., null mutations in LPL, GPIHBP1, APOC2, APOA5, or LMF1) that typically result in extremely high TG levels exceeding 900 mg / dL. As shown in the examples herein, the APOC3 RNAi drug substances listed in Table 2 provide substantial TG-reducing effects suitable for the treatment of diseases and disorders.

[0116] Drug administration and inhibition of APOC3 gene expression Generally, the effective dose of an APOC3 RNAi agent is in the range of approximately 0.1 to 10 mg per kg of body weight per dose, for example, approximately 0.25 to 5 mg per kg of body weight per dose. In some embodiments, the effective dose of an APOC3 RNAi agent is in the range of approximately 0.5 to 4 mg per kg of body weight per dose. In some embodiments, the effective dose is a fixed dose. In some embodiments, a fixed dose of an APOC3 RNAi drug substance between 1 mg and 100 mg is the effective dose. In some embodiments, a fixed dose of an APOC3 RNAi drug substance between 10 mg and 50 mg is the effective dose. In some embodiments, a fixed dose of an APOC3 RNAi drug substance between 20 mg and 50 mg is the effective dose. In some embodiments, a fixed dose of an APOC3 RNAi drug substance between 25 mg and 50 mg is the effective dose. The amount administered is likely to depend on variables such as the patient's overall age and health status, the relative bioefficacy 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 approximately 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 approximately 10 mg, approximately 25 mg, approximately 50 mg, or approximately 100 mg is an effective dose.

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

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

[0119] In some embodiments, the APOC3 mRNA levels of subjects administered with the 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 more than 99%. In some embodiments, APOC3 mRNA levels in subjects administered with the described APOC3 RNAi agent are reduced by at least approximately 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 subjects before administration of the APOC3 RNAi agent or subjects that have not received the APOC3 RNAi agent. Gene expression levels and / or mRNA levels in the subject are reduced in the subject's cells, cell populations, and / or tissues.

[0120] In some embodiments, APOC3 protein levels in subjects administered with the 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 more than 99% compared to subjects before administration of the APOC3 RNAi agent or subjects not receiving the APOC3 RNAi agent. In some embodiments, APOC3 protein levels in subjects administered with the described APOC3 RNAi agent are 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 subjects before administration of the APOC3 RNAi agent or subjects not receiving the APOC3 RNAi agent. Protein levels in the subject may be reduced in the subject's cells, cell populations, tissues, blood, and / or other bodily fluids.

[0121] Reductions in APOC3 gene expression, APOC3 mRNA, or APOC3 protein levels can be evaluated and quantified by common methods known in the art. The examples disclosed herein generally illustrate known methods for evaluating inhibition of APOC3 gene expression and reductions in APOC3 protein levels. Reductions or decreases in APOC3 mRNA and / or protein levels are collectively referred herein to as reductions or decreases in APOC3, or inhibition or reduction of APOC3 expression.

[0122] In some embodiments, triglyceride levels in subjects administered with the described APOC3 RNAi agent are reduced by at least approximately 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 subjects before administration of the APOC3 RNAi agent or subjects not receiving the APOC3 RNAi agent. Triglyceride levels in subjects are reduced in the subject's blood / serum.

[0123] In some embodiments, very low-density lipoprotein cholesterol (VLDL-C) levels in subjects administered with the described APOC3 RNAi agent are reduced by at least approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or more than 85% compared to subjects before or without the APOC3 RNAi agent. VLDL-C levels in subjects are reduced in the subject's blood / serum.

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

[0125] In some embodiments, high-density lipoprotein cholesterol (HDL-C) levels in subjects administered with the described APOC3 RNAi agent are increased by at least approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or more than 85% compared to subjects before administration of the APOC3 RNAi agent or subjects not receiving the APOC3 RNAi agent. Total HDL-C levels in the subjects are increased in the subjects' cells, cell populations, tissues, blood, and / or other bodily fluids.

[0126] In some embodiments, apolipoprotein A1 (ApoA1) levels in subjects administered with the described APOC3 RNAi agent are 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 subjects before administration of the APOC3 RNAi agent or subjects not receiving the APOC3 RNAi agent. ApoA1 levels in subjects are reduced in the cells, cell populations, tissues, blood, and / or other bodily fluids of the subjects.

[0127] As used herein, the terms “to treat,” “treatment,” etc., mean a method or step taken to provide relief or mitigation of one or more symptoms, severity, and / or frequency of a disease in a subject. As used herein, “to treat” and “treatment” may include prevention, control, preventive measures, and / or inhibition of one or more symptoms, severity, and / or frequency of a disease in a subject. As used herein, unless otherwise expressly noted, the phrase “treatment of disease or disorder” includes treatment of the underlying condition and / or symptoms of the disease, as understood by those skilled in the art. Unless otherwise expressly noted herein, such phrase is not intended to be limiting.

[0128] As used herein, “APOC3-related disease or disorder” means any disease or disorder that can be treated at least in part by reduction or inhibition of APOC3 gene expression using APOC3 RNAi agents, including by the methods of treatment described herein. Where discussed herein, the methods of treatment disclosed herein, among other things, result in a reduction in TG levels in subjects administered with the APOC3 RNAi drug substance. 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 mellitus, lipodystrophy syndromes including familial partial lipodystrophy, obesity, dyslipidemia, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hyperlipidemia, hypertriglyceridemia, lipid and / or cholesterol metabolism disorders, atherosclerosis, cardiovascular disease, coronary artery disease, and other dyslipidemia and metabolism-related disorders and diseases.

[0129] As used herein, “monthly dosing” or “monthly” administration means every 28 days. As used herein, “quarterly dosing” or “quarterly” administration means every 84 days. The term “about” when used in conjunction with monthly dosing means monthly dosing + / - 5 days. The term “about” when used in conjunction with quarterly dosing means quarterly dosing + / - 14 days.

[0130] As used herein, the phrase “introduced into cells” means functionally delivering an RNAi agent into cells, when referring to an RNAi agent. The phrase “functional delivery” means delivering an RNAi agent to cells in a manner that enables the RNAi agent to have its expected biological activity, for example, sequence-specific inhibition of gene expression.

[0131] Unless otherwise stated, symbols [ka] The use of means, when used herein, means that any base(s) or any other base(s) that conform to the scope of the inventions described herein may be linked thereto.

[0132] Where used herein, unless specifically identified by a structure having a particular conformation, each structure disclosed herein is intended to represent all such possible isomers, including their optically pure and racemic forms, for each structure that contains a chiral center and thus results in enantiomers, diastereomers, or other stereoisomer configurations. 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 component not specified in the claims. As used in the claims herein, the phrase "essentially consisting of" limits the claims to those that do not substantially affect the specified material or step, and the fundamental and novel features(s) of the claimed invention.

[0134] Those skilled in the art will readily understand and recognize 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. Therefore, when used herein, the structures disclosed herein assume that certain functional groups, such as OH, SH, or NH, may be protonated or deprotonated. The disclosure herein is intended to cover the disclosed compounds and compositions regardless of their protonation state 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 those generally understood by those skilled in the art. Methods and materials similar to or equivalent to those described herein may 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 referenced herein are thus incorporated herein by reference in their entirety. In case of any conflict, this specification, including definitions, shall prevail. Furthermore, materials, methods and examples are illustrative and not intended to be limiting.

[0136] The embodiments and items provided above are illustrated herein by the following non-limiting embodiments. [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 is known in the art. Commercial oligonucleotide synthesizers (e.g., MerMade96E® (Bioautomation) or MerMade12® (Bioautomation)) may be used. Synthesis may be carried out on a solid support made of controlled pore glass (CPG, 500 Å or 600 Å, obtained from Prime Synthesis, Aston, PA, USA). Monomers located at the 3' end of each strand may be bound to the solid support as a starting point for synthesis. All RNAs, 2'-modified RNA phosphoramidites, and inverted debasic phosphoramidites can be purchased commercially. Targeting group-containing phosphoramidites suitable for addition to the 5' end of the sense strand can be synthesized. Standard cleavage, deprotection, purification, and annealing steps may be used, as is known in the art. Further descriptions relating to 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 thus incorporated herein by reference in whole. The APOC3 RNAi drug substances can then be formulated by dissolving them in standard pharmaceutically acceptable excipients commonly 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 APOC3 RNAi drug (ADS-005) in healthy human volunteers and patients with severe hypertriglyceridemia and familial chylomicronemia syndrome (FCS). Phase 1 single-dose and multi-dose escalation studies were initiated to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamic effects of the APOC3 RNAi drug (ADS-005) in healthy adult volunteers and patients with severe hypertriglyceridemia and FCS. The study subject population was 19.0 kg / m². 2 and 40.0 kg / m 2 This includes adult men and women aged 18-65 with a BMI between the following, and • Cohorts 1, 2, 3, and 4: All subjects had fasting triglycerides >80 mg / dL (>0.903 mmol / L) at screening and had not received any lipid-lowering or triglyceride-lowering treatments. Each double-blind cohort enrolled 10 subjects (6 receiving the active drug; 4 receiving placebo (PBO)), and all cohorts received a single escalating dose of APOC3 RNAi drug substance or PBO at escalating dose levels of 10, 25, 50, and 100 mg, as shown in Figures 4 and 5. • Cohorts 1b, 2b, 3b, and 4b: Each cohort will be double-blinded, with 8 active drug patients and 2 placebo patients per cohort, having a fasting serum triglyceride level of at least 300 mg / dL (3.38 mmol / L) at screening. All subjects will receive multiple escalating doses of either the APOC3 RNAi drug substance or placebo at dose levels of 10, 25, 50, or 100 mg. • Cohort 5: The cohort is open-label and includes up to eight patients with chylomicronemia. Each subject in Cohort 5 receives two 50 mg doses of APOC3 RNAi drug. • Cohorts 6, 7, and 8: All subjects had fasting triglycerides >80 mg / dL (>0.903 mmol / L) at screening and had not received any lipid-lowering or triglyceride-lowering treatments. Each double-blind cohort enrolled 10 subjects (6 receiving the active drug; 4 receiving placebo (PBO)), and all cohorts received a single escalating dose of APOC3 RNAi drug substance or PBO at escalating dose levels of 10, 25, 50, and 100 mg, as shown in Figures 4 and 5.

[0139] Subjects were separated into various cohorts based on the criteria discussed above (see also Figure 5). Cohorts 1-4 were randomized to receive either an APOC3 RNAi drug or a placebo, administered subcutaneously in a single dose of 25 mg (Cohort 1), 50 mg (Cohort 2), 100 mg (Cohort 3), or 10 mg (Cohort 4) (6 received the active drug; 4 received placebo). The other cohorts received two doses via subcutaneous injection: 10 mg (Cohorts 1b and 6), 50 mg (Cohorts 2b and 8), 100 mg (Cohort 3b), or 25 mg (Cohorts 4b and 7) (administered on days 1 and 29). In addition, two doses of 50 mg were administered to patients with chylomicronemia (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. The study parameters are summarized in Table 4 below.

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

[0141] Initially, the cohort was proposed as part of the clinical trial protocol with an APOC3 RNAi drug at a dose of 200 mg per dose. However, considering the unexpected and remarkable 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 administered at a lower dose of 10 mg. All doses—10 mg, 25 mg, 50 mg, and 100 mg—resulted in substantial serum APOC3 reduction. For example, both the 25 mg and 50 mg cohorts reached a mean serum APOC3 protein reduction of approximately 85% after the single dose in the Phase I study by day 22, and the 100 mg cohort reached a mean serum APOC3 protein reduction of approximately 85% after the single dose in the Phase I study by day 8. Figures 6–9 report on the serum APOC3 reductions of cohorts 1, 2, 3, and 4 in the Phase I study.

[0142] The serum APOC3 reduction results from the study indicate that administration of APOC3 RNAi drugs at doses of at least 10 mg, 25 mg, 50 mg, and 100 mg resulted in a deep and sustained reduction of serum APOC3 compared to placebo. As noted herein, APOC3 is primarily synthesized in hepatocytes in the liver, but the data suggest that as much as 20% of APOC3 protein can be expressed in intestinal cells in the gastrointestinal tract. Surprisingly and unexpectedly, a dose level of as little as 25 mg of APOC3 RNAi drug resulted in a substantial knockdown of APOC3 gene expression reaching approximately 85% (similar to the knockdown levels observed at higher doses of 50 mg and 100 mg), indicating the achievement of almost complete suppression of APOC3 expressed in hepatocytes. Also unexpectedly, a dose level of as little as 10 mg of APOC3 RNAi drug resulted in a substantial knockdown, showing a reduction of up to approximately 72% of APOC3 gene expression compared to baseline.

[0143] The duration of the serum APOC3 reduction (>80%) from a single dose of 25 mg also appears to be unexpectedly persistent. Up to 85 days after dose administration, an APOC3 reduction of ≥70% was observed at day 113 (end of the study). Even at the 10 mg dose, the duration of effect was long, with an APOC3 reduction of >60% observed up to 57 days after dose administration. These data suggest that monthly, quarterly, or even every six months dosing of 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 various cohorts after administration of 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) groups achieved a maximum TG reduction of approximately 60% compared to baseline. Up to day 113 (end of the study), both the 25 mg (cohort 1) and 50 mg (cohort 2) groups maintained a reduction of approximately 50% in serum TG. Furthermore, the 10 mg (cohort 4) group achieved a maximum TG reduction of approximately 50% and maintained a reduction of approximately 40% in serum TG at day 99.

[0145] A reduction in VLDL-C, similar to that observed in serum triglycerides (TG), was also observed. For example, on day 29, the 25 mg (cohort 1), 50 mg (cohort 2), and 100 mg (cohort 3) groups each achieved approximately 60% reductions in VLDL-C levels, while the 10 mg group (cohort 4) reported approximately 50% reductions on day 29. Furthermore, the data showed approximately 10%–20% reductions in total cholesterol and low-density lipoprotein cholesterol (LDL-C) in the subjects.

[0146] Furthermore, 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 Phase 1 studies, it has been observed that the increase in HDL-C is slightly greater than that seen in APOC3-deficient individuals (homozygous or heterozygous).

[0147] Multiple dose data from patients in Cohort 5 (n=5, all chylomicronemia patients with data up to day 29) also showed reductions in serum APOC3 protein levels and lipid parameters. Regarding APOC3 protein levels, a maximum mean reduction of 96% (-62 mg / dL) was observed across three subjects at a mean baseline APOC3 level of 66 mg / dL. A maximum mean reduction of 92% (-2583 mg / dL) in triglycerides was observed across these five subjects at a mean baseline value of 2787 mg / dL ± 1444 mg / dL (standard deviation). In one subject, a maximum individual reduction of 99% (-86.7 mg / dL) was observed after a single dose at a starting concentration of 88 mg / dL. Furthermore, 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 level 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, the Phase I study has reported no deaths, no serious adverse events (SAEs), and no adverse events (AEs) classified as severe. Most AEs have been reported as mild.

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

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

[0151] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of: administering to the subject a pharmaceutical composition containing an APOC3 RNAi agent in a first dose between approximately 1 mg and approximately 100 mg of the APOC3 RNAi agent; and administering to the subject a second dose between approximately 1 mg and approximately 100 mg of the pharmaceutical composition containing an APOC3 RNAi agent in a second dose between approximately 4 months after the first dose, wherein the first and second doses are administered by subcutaneous injection.

[0152] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of: administering to the subject a pharmaceutical composition containing an APOC3 RNAi agent in a first dose between approximately 1 mg and approximately 100 mg of the APOC3 RNAi agent; and administering to the subject a second dose between approximately 1 mg and approximately 100 mg of the pharmaceutical composition containing an APOC3 RNAi agent in a second dose between approximately 1 mg and approximately 100 mg, about six months after the first dose, wherein the first and second doses are administered by subcutaneous injection.

[0153] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering a first dose of a pharmaceutical composition containing an APOC3 RNAi agent in a dose of APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg to a human subject by subcutaneous administration. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately one month after the first dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 3 to 6 months after the second dose. Methods including the above are disclosed herein.

[0154] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering a first dose of a pharmaceutical composition containing an APOC3 RNAi agent, in a dose of the APOC3 RNAi agent between approximately 1 mg and approximately 100 mg, to a human subject by subcutaneous administration. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately one month after the first dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 6 months after the second dose. Methods including the above are disclosed herein.

[0155] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering a first dose of a pharmaceutical composition containing an APOC3 RNAi agent, in a dose of the APOC3 RNAi agent between approximately 1 mg and approximately 100 mg, to a human subject by subcutaneous administration. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately 1 to 6 months after the first dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 3 months (e.g., 4 doses per year or every 12 weeks (q12w)) to approximately 6 months after the second dose. Methods including the above are disclosed herein.

[0156] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering a first dose of a pharmaceutical composition containing an APOC3 RNAi agent in a dose of APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg to a human subject by subcutaneous administration. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately 3 to 6 months after the first dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 6 months after the second dose. Methods including the above are disclosed herein.

[0157] In some embodiments, the methods disclosed herein involve each dose being between approximately 10 mg and approximately 100 mg of APOC3 RNAi agent.

[0158] In some embodiments, the methods disclosed herein involve each dose being between approximately 10 mg and approximately 50 mg of APOC3 RNAi agent.

[0159] In some embodiments, the methods disclosed herein involve each dose being between approximately 25 mg and approximately 100 mg of APOC3 RNAi agent.

[0160] In some embodiments, the methods disclosed herein involve each dose being between approximately 25 mg and approximately 50 mg of APOC3 RNAi agent.

[0161] In some embodiments, the methods disclosed herein involve a dose of approximately 10 mg of APOC3 RNAi agent.

[0162] In some embodiments, the methods disclosed herein involve a dose of approximately 25 mg of each APOC3 RNAi agent.

[0163] In some embodiments, the methods disclosed herein involve a dose of approximately 50 mg of the APOC3 RNAi agent.

[0164] In some embodiments, the methods disclosed herein involve a dose of approximately 100 mg of APOC3 RNAi agent.

[0165] In some embodiments, the methods disclosed herein involve administering further therapeutic agents to the subject for the treatment of APOC3-related disease or disorder.

[0166] In some embodiments, the methods disclosed herein refer to APOC3-related disorders or conditions as dyslipidemia.

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

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

[0169] In some embodiments, the methods disclosed herein define APOC3-related disorder or impairment as hypertriglyceridemia with or without a history of pancreatitis.

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

[0171] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of: administering to the subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg; and administering to the subject a second dose of the pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose between approximately 1 mg and approximately 100 mg, approximately one month after the initial dose, wherein the initial dose and the second dose are administered by subcutaneous injection.

[0172] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of: administering to the subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg; and administering to the subject a second dose of the pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose between approximately 1 mg and approximately 100 mg, approximately three months after the initial dose (for example, four doses per year or every 12 weeks (q12w)), wherein the initial and second doses are administered by subcutaneous injection.

[0173] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of: administering to the subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg; and administering to the subject a second dose of the pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose between approximately 1 mg and approximately 100 mg, approximately 4 months after the initial dose, wherein the initial and second doses are administered by subcutaneous injection.

[0174] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of: administering to the subject a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg; and administering to the subject a second dose of the pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2 in an initial dose of approximately 1 mg and approximately 100 mg, approximately 6 months after the initial dose, wherein the initial and second doses are administered by subcutaneous injection.

[0175] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering to a human subject by subcutaneous administration an initial dose of a pharmaceutical composition containing an APOC3 RNAi drug substance, with dosages ranging from approximately 1 mg to approximately 100 mg as listed in Table 2. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately one month after the initial dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 3 to 6 months after the second dose. Methods including the above are disclosed herein.

[0176] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering to a human subject by subcutaneous administration an initial dose of a pharmaceutical composition containing an APOC3 RNAi drug substance, with dosages ranging from approximately 1 mg to approximately 100 mg as listed in Table 2. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately one month after the initial dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 6 months after the second dose. Methods including the above are disclosed herein.

[0177] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering to a human subject by subcutaneous administration an initial dose of a pharmaceutical composition containing an APOC3 RNAi drug substance, with dosages ranging from approximately 1 mg to approximately 100 mg as listed in Table 2. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately 1 to 6 months after the initial dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 3 months (e.g., 4 doses per year or every 12 weeks (q12w)) to approximately 6 months after the second dose. Methods including the above are disclosed herein.

[0178] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering to a human subject by subcutaneous administration an initial dose of a pharmaceutical composition containing an APOC3 RNAi drug substance, with dosages ranging from approximately 1 mg to approximately 100 mg as listed in Table 2. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately 3 to 6 months after the initial dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 6 months after the second dose. Methods including the above are disclosed herein.

[0179] In some embodiments, the methods disclosed herein involve each dose being an APOC3 RNAi drug substance between approximately 10 mg and approximately 100 mg.

[0180] In some embodiments, the methods disclosed herein involve each dose being an APOC3 RNAi drug substance between approximately 10 mg and approximately 50 mg.

[0181] In some embodiments, the methods disclosed herein involve each dose being an APOC3 RNAi drug substance between approximately 25 mg and approximately 100 mg.

[0182] In some embodiments, the methods disclosed herein involve each dose being an APOC3 RNAi drug substance between approximately 25 mg and approximately 50 mg.

[0183] In some embodiments, the methods disclosed herein involve a dose of approximately 10 mg of the APOC3 RNAi drug substance.

[0184] In some embodiments, the methods disclosed herein involve a dose of approximately 25 mg of the APOC3 RNAi drug substance.

[0185] In some embodiments, the methods disclosed herein involve a dose of approximately 50 mg of the APOC3 RNAi drug substance.

[0186] In some embodiments, the methods disclosed herein involve a dose of approximately 100 mg of the APOC3 RNAi drug substance.

[0187] In some embodiments, the methods disclosed herein involve a pharmaceutical composition comprising, consisting of, or essentially consisting of, a formulated APOC3 RNAi drug substance as listed in Table 3.

[0188] In some embodiments, the method disclosed herein further includes a step of administering a further dose after the first dose, the further dose being administered at intervals of about one month, about three months (for example, every 12 weeks, or once every calendar quarter), about four months, or about six months.

[0189] In some embodiments, the method disclosed herein further includes the step of administering a further dose after a second dose, the further dose being administered at intervals of about one month, about three months (for example, every 12 weeks, or once every calendar quarter), about four months, or about six months.

[0190] In some embodiments, the method disclosed herein further includes a step of administering a further dose after a third dose, the further dose being administered at intervals of about one month, about three months (for example, every 12 weeks, or once every calendar quarter), about four months, or about six months.

[0191] In some embodiments, the method disclosed herein further includes the step of administering additional doses at intervals of about 3 to about 6 months.

[0192] In some embodiments, a method for inhibiting APOC3 expression in subjects requiring inhibition of APOC3 expression is disclosed herein, comprising the step of administering to the subject one or more doses of the APOC3 RNAi drug substances listed in Table 2, in amounts ranging from about 1 mg to about 100 mg, over a period of about one month.

[0193] In some embodiments, a method for inhibiting APOC3 expression in subjects requiring inhibition of APOC3 expression is disclosed herein, comprising the step of administering to the subject one or more doses of the APOC3 RNAi drug substances listed in Table 2, in amounts ranging from about 1 mg to about 100 mg, over a period of about 3 months.

[0194] In some embodiments, the methods disclosed herein involve a dose of an APOC3 RNAi drug substance between approximately 10 mg and approximately 100 mg.

[0195] In some embodiments, the methods disclosed herein involve a dose of an APOC3 RNAi drug substance between approximately 10 mg and approximately 50 mg.

[0196] In some embodiments, the methods disclosed herein involve a dose of an APOC3 RNAi drug substance between approximately 25 mg and approximately 100 mg.

[0197] In some embodiments, the methods disclosed herein involve a dose of an APOC3 RNAi drug substance between approximately 25 mg and approximately 50 mg.

[0198] In some embodiments, the method disclosed herein involves a dose of approximately 10 mg of APOC3 RNAi drug substance.

[0199] In some embodiments, the method disclosed herein involves a dose of approximately 25 mg of APOC3 RNAi drug substance.

[0200] In some embodiments, the method disclosed herein involves a dose of approximately 50 mg of APOC3 RNAi drug substance.

[0201] In some embodiments, the method disclosed herein involves a dose of approximately 100 mg of APOC3 RNAi drug substance.

[0202] In some embodiments, a pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2, preferably in a pharmaceutically acceptable salt form, and APOC3 The RNAi drug substance comprises an antisense strand containing the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand containing the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasic deoxyribose residue; (NAG37)s represents [ka] A pharmaceutical composition comprising the structure shown herein, wherein the pharmaceutical composition is administered to a subject in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg, and the pharmaceutical composition is administered to the subject in a second dose of an APOC3 RNAi drug substance listed in Table 2 between approximately 1 mg and approximately 100 mg, approximately one month after the initial dose, and the initial and second doses are administered by subcutaneous injection, is disclosed herein.

[0203] In some embodiments, a pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2, preferably in a pharmaceutically acceptable salt form, and APOC3 The RNAi drug substance comprises an antisense strand containing the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand containing the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasic deoxyribose residue; (NAG37)s represents [ka] A pharmaceutical composition comprising the structure shown herein is disclosed herein, wherein the pharmaceutical composition is administered to a subject in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg, and the pharmaceutical composition is administered to the subject in a second dose of an APOC3 RNAi drug substance listed in Table 2 between approximately 1 mg and approximately 100 mg, approximately 3 months after the initial dose, and the initial and second doses are administered by subcutaneous injection.

[0204] In some embodiments, a pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2, preferably in a pharmaceutically acceptable salt form, and APOC3 The RNAi drug substance comprises an antisense strand containing the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand containing the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasic deoxyribose residue; (NAG37)s represents [ka] A pharmaceutical composition comprising the structure shown herein is disclosed herein, wherein the pharmaceutical composition is administered to a subject in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg, and the pharmaceutical composition is administered to the subject in a second dose of an APOC3 RNAi drug substance listed in Table 2 between approximately 1 mg and approximately 100 mg, approximately 4 months after the initial dose, and the initial and second doses are administered by subcutaneous injection.

[0205] In some embodiments, a pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance listed in Table 2, preferably in a pharmaceutically acceptable salt form, and APOC3 The RNAi drug substance comprises an antisense strand containing the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand containing the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasic deoxyribose residue; (NAG37)s represents [ka] A pharmaceutical composition comprising the structure shown herein is disclosed herein, wherein the pharmaceutical composition is administered to a subject in an initial dose of an APOC3 RNAi drug substance between approximately 1 mg and approximately 100 mg, and the pharmaceutical composition is administered to the subject in a second dose of an APOC3 RNAi drug substance listed in Table 2 between approximately 1 mg and approximately 100 mg, approximately 6 months after the initial dose, and the initial and second doses are administered by subcutaneous injection.

[0206] In some embodiments, a pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance as described in Table 2. a. The pharmaceutical composition was administered to human subjects in an initial dose of APOC3 RNAi drug substance between approximately 1 mg and 100 mg. b. The pharmaceutical composition was administered to human subjects at a second dose of the APOC3 RNAi drug substance, between approximately 1 mg and 100 mg, approximately one month after the initial dose. c. The pharmaceutical composition is administered to human subjects at a third dose of APOC3 RNAi drug substance between approximately 1 mg and 100 mg, approximately 3 to 6 months after the second dose. Pharmaceutical compositions are disclosed herein.

[0207] In some embodiments, a pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance as described in Table 2. a. The pharmaceutical composition was administered to human subjects in an initial dose of APOC3 RNAi drug substance between approximately 1 mg and 100 mg. b. The pharmaceutical composition was administered to human subjects at a second dose of the APOC3 RNAi drug substance, between approximately 1 mg and 100 mg, approximately one month after the initial dose. c. The pharmaceutical composition is administered to human subjects at a third dose of the APOC3 RNAi drug substance, between approximately 1 mg and 100 mg, approximately 6 months after the second dose. Pharmaceutical compositions are disclosed herein.

[0208] In some embodiments, a pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance as described in Table 2. a. The pharmaceutical composition was administered to human subjects in an initial dose of APOC3 RNAi drug substance between approximately 1 mg and 100 mg. b. The pharmaceutical composition was administered to human subjects at a second dose of the APOC3 RNAi drug substance, between approximately 1 mg and 100 mg, approximately 1 to 6 months after the initial dose. c. The pharmaceutical composition is administered to human subjects at a third dose of APOC3 RNAi drug substance between approximately 1 mg and 100 mg, approximately 3 to 6 months after the second dose. Pharmaceutical compositions are disclosed herein.

[0209] A pharmaceutical composition for use in treating APOC3-related disease or disorder in human subjects requiring treatment of APOC3-related disease or disorder, wherein the use comprises the administration of a pharmaceutical composition containing an APOC3 RNAi drug substance as described in Table 2. a. The pharmaceutical composition was administered to human subjects in an initial dose of APOC3 RNAi drug substance between approximately 1 mg and 100 mg. b. The pharmaceutical composition was administered to human subjects at a second dose of the APOC3 RNAi drug substance, between approximately 1 mg and 100 mg, approximately 3 to 6 months after the initial dose. c. The pharmaceutical composition is administered to human subjects at a third dose of the APOC3 RNAi drug substance, between approximately 1 mg and 100 mg, approximately 6 months after the second dose. Pharmaceutical composition.

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

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

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

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

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

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

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

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

[0218] In some embodiments, the pharmaceutical composition, as used as disclosed herein, is administered to a human subject together with further therapeutic agents for the treatment of APOC3-related diseases or disorders.

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

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

[0221] In some embodiments, the pharmaceutical composition as used as disclosed herein is used to treat familial chylomicronemia syndrome (FCS), chylomicronemia or multiple factor chylomicronemia.

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

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

[0224] In some embodiments, the pharmaceutical composition as used as disclosed herein comprises, consists of or consists essentially of the formulated APOC3 RNAi drug substance described in Table 3.

[0225] In some embodiments, the pharmaceutical composition as used as disclosed herein is administered to a subject in a further dose administered about one month apart.

[0226] In some embodiments, the pharmaceutical composition as used as disclosed herein is administered to a subject in a further dose administered about three months apart.

[0227] In some embodiments, the pharmaceutical composition as used as disclosed herein is administered to a subject in a further dose administered about four months apart.

[0228] In some embodiments, the pharmaceutical composition as used as disclosed herein is administered to a subject in a further dose administered about six months apart.

[0229] In some embodiments, the pharmaceutical composition as used as disclosed herein is administered to a subject in a further dose administered about three to about six months apart.

[0230] In some embodiments, the pharmaceutical composition, as used as disclosed herein, is self-administered by the subject.

[0231] In some embodiments, the pharmaceutical composition, as used as disclosed herein, is self-administered by a healthcare professional.

[0232] In some embodiments, the pharmaceutical compositions, as used as disclosed herein, reduce serum APOC3 protein levels in human subjects by more than 60% compared to baseline levels.

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

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

[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 human subjects compared to baseline levels.

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

[0237] In some embodiments, the pharmaceutical composition, as used as disclosed herein, comprises an APOC3 RNAi drug substance in the form of a pharmaceutically acceptable sodium salt.

[0238] In some embodiments, pharmaceutical compositions are disclosed herein for inhibiting APOC3 expression in human subjects requiring inhibition of APOC3 expression, wherein the pharmaceutical composition comprises one or more doses of the APOC3 RNAi drug substances listed in Table 2, in amounts of about 1 mg to about 100 mg, administered over a period of about one month.

[0239] In some embodiments, pharmaceutical compositions are disclosed herein for inhibiting APOC3 expression in human subjects requiring inhibition of APOC3 expression by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance listed in Table 2, wherein the pharmaceutical composition is administered over a period of about 3 months in doses of one or more APOC3 RNAi drug substances listed in Table 2 in amounts of about 1 mg to about 100 mg.

[0240] In some embodiments, pharmaceutical compositions are disclosed herein for inhibiting APOC3 expression in human subjects requiring inhibition of APOC3 expression by administration of a pharmaceutical composition comprising an APOC3 RNAi drug substance listed in Table 2, wherein the pharmaceutical composition is administered over a period of about one month in doses of one or more APOC3 RNAi drug substances listed in Table 2 in amounts of about 10 mg to about 100 mg.

[0241] In some embodiments, pharmaceutical compositions are disclosed herein for inhibiting APOC3 expression in human subjects requiring inhibition of APOC3 expression, wherein the pharmaceutical composition comprises one or more doses of the APOC3 RNAi drug substances listed in Table 2, in amounts of about 10 mg to about 100 mg, administered over a period of about 3 months.

[0242] In some embodiments, a pharmaceutical composition for inhibiting the expression of APOC3 in a human subject that requires inhibition of the expression of APOC3, by administration of a pharmaceutical composition comprising the APOC3 RNAi drug substance described in Table 2, wherein 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 are administered over a period of about 6 months, is disclosed herein.

[0243] In some embodiments, a pharmaceutical composition for inhibiting the expression of APOC3 in a human subject that requires inhibition of the expression of APOC3, by administration of a pharmaceutical composition comprising the APOC3 RNAi drug substance described in Table 2, wherein one or more doses of the APOC3 RNAi drug substance described in Table 2 in an amount of about 25 mg to about 50 mg are administered over a period of about 1 month, is disclosed herein.

[0244] In some embodiments, a pharmaceutical composition for inhibiting the expression of APOC3 in a human subject that requires inhibition of the expression of APOC3, by administration of a pharmaceutical composition comprising the APOC3 RNAi drug substance described in Table 2, wherein one or more doses of the APOC3 RNAi drug substance described in Table 2 in an amount of about 25 mg to about 50 mg are administered over a period of about 3 months, is disclosed herein.

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

[0246] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of administering to the subject a pharmaceutical composition containing an APOC3 RNAi drug substance as listed in Table 2 for doses of approximately 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 the doses are administered by subcutaneous injection.

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

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

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

[0250] In some embodiments, a method for treating an APOC3-related disease or disorder in a human subject requiring treatment of such disease or disorder is disclosed herein, comprising the steps of administering to the subject a pharmaceutical composition containing an APOC3 RNAi drug substance as listed in Table 2 for doses of approximately 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 the doses are administered by subcutaneous injection.

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

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

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

[0254] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering a dose of a pharmaceutical composition containing an APOC3 RNAi drug substance of approximately 100 mg or less to a human subject. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately 1 to 6 months after the initial dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 6 months after the second dose. A method of administration, including the use of a substance and administered by subcutaneous injection, is disclosed herein.

[0255] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering a dose of a pharmaceutical composition containing an APOC3 RNAi drug substance of approximately 100 mg or less to a human subject. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately one month after the initial dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 3 months (e.g., once every calendar quarter or every 12 weeks (q12w)) to approximately 6 months after the second dose. A method of administration, including the use of a substance and administered by subcutaneous injection, is disclosed herein.

[0256] In some embodiments, a method for treating APOC3-related disease or disorder in a human subject requiring treatment for APOC3-related disease or disorder, a. A step of administering a dose of a pharmaceutical composition containing an APOC3 RNAi drug substance of approximately 100 mg or less to a human subject. b. A step of administering a second dose of the pharmaceutical composition to a human subject approximately 3 months (e.g., once every calendar quarter or every 12 weeks (q12w)) to approximately 6 months after the initial dose, and c. A step in which a third dose of the pharmaceutical composition is administered to a human subject approximately 6 months after the second dose. A method of administration, including the use of a substance and administered by subcutaneous injection, is disclosed herein.

[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, and 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 for treating APOC3-related disease or disorder in a human subject requiring treatment for such disease or disorder, comprising the steps of administering to the subject a pharmaceutical composition comprising, preferably, a pharmaceutically acceptable salt form of APOC3 RNAi drug substance, the APOC3 The RNAi drug substance comprises an antisense strand containing the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand containing the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasic deoxyribose residue; (NAG37)s represents [ka] A method comprising the steps of administering a first dose of the APOC3 RNAi drug substance, comprising a structure shown by, between approximately 1 mg and approximately 100 mg, to the subject, and administering a second dose of the pharmaceutical composition comprising the APOC3 RNAi drug substance, between approximately 1 mg and approximately 100 mg, to the subject about one month after the first dose, wherein the first and second doses are administered by subcutaneous injection. (Item 2) A method for treating APOC3-related disease or disorder in a human subject requiring treatment for such disease or disorder, comprising the steps of administering to the subject a pharmaceutical composition comprising, preferably, a pharmaceutically acceptable salt form of APOC3 RNAi drug substance, the APOC3 The RNAi drug substance comprises an antisense strand containing the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand containing the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasic deoxyribose residue; (NAG37)s represents [ka] A method comprising the steps of: administering the APOC3 RNAi drug substance, comprising the structure shown by, in a first dose between approximately 1 mg and approximately 100 mg of the APOC3 RNAi drug substance; and administering to the subject a second dose of the pharmaceutical composition comprising the APOC3 RNAi drug substance, between approximately 1 mg and approximately 100 mg, about three months after the first dose, wherein the first and second doses are administered by subcutaneous injection. (Item 3) A method for treating APOC3-related disease or disorder in a human subject requiring treatment for such disease or disorder, comprising the steps of administering to the subject a pharmaceutical composition comprising, preferably, a pharmaceutically acceptable salt form of APOC3 RNAi drug substance, the APOC3 The RNAi drug substance comprises an antisense strand containing the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand containing the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a, c, g, i, and u represent 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine, respectively; Af, Cf, Gf, and Uf represent 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted debasic deoxyribose residue; (NAG37)s represents

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Claims

1. A pharmaceutical composition for use in treating an APOC3-related disease or disorder in a human subject requiring treatment for such disease or disorder, wherein the use comprises the step of administering to the subject a pharmaceutical composition comprising an APOC3 RNAi drug substance, preferably in a pharmaceutically acceptable salt form, wherein the APOC3 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), wherein a, c, g, i, and u are 2'-O-methyladenosine, 2'- O-methylcytidine, 2'-O-methylguanosine, 2'-O-methylinosine, and 2'-O-methyluridine are represented; Af, Cf, Gf, and Uf are represented by 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s indicates a phosphorothioate linkage; (invAb) indicates an inverted debasic deoxyribose residue; (NAG37)s is, 【Chemistry 9】 A pharmaceutical composition comprising the steps of: administering the APOC3 RNAi drug substance in a first dose between approximately 1 mg and approximately 100 mg, wherein the APOC3 RNAi drug substance comprises the steps of: administering the subject a second dose of the pharmaceutical composition comprising the APOC3 RNAi drug substance in a second dose between approximately 1 mg and approximately 100 mg, approximately three months after the first dose, wherein the first and second doses are administered by subcutaneous injection.

2. The pharmaceutical composition according to claim 1, further comprising the step of administering a further dose after the second dose, wherein the further dose is administered at intervals of approximately three months.

3. The pharmaceutical composition according to any one of claims 1 to 2, wherein the third dose is administered approximately three months after the second dose.

4. The pharmaceutical composition according to claim 3, further comprising the step of administering a further dose after the third dose, wherein the further dose is administered at intervals of approximately 3 to 6 months.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein each dose is the APOC3 RNAi drug substance between approximately 10 mg and approximately 100 mg.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein each dose is the APOC3 RNAi drug substance between approximately 25 mg and approximately 50 mg.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein each dose of the APOC3 RNAi drug substance is approximately 25 mg.

8. The pharmaceutical composition according to any one of claims 1 to 6, wherein each dose of the APOC3 RNAi drug substance is approximately 50 mg.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the subject is further administered a further therapeutic agent for the treatment of APOC3-related disease or disorder.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the APOC3-related disease or disorder is dyslipidemia.

11. The pharmaceutical composition according to any one of claims 1 to 9, wherein the APOC3-related disease or disorder is hypertriglyceridemia, obesity, dyslipidemia, non-alcoholic fatty liver disease, hyperlipidemia, lipid and / or cholesterol metabolism disorder, 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.

12. The pharmaceutical composition according to any one of claims 1 to 9, wherein the APOC3-related disease or disorder is chylomicronemia.

13. The pharmaceutical composition according to any one of claims 1 to 9, wherein the APOC3-related disease or disorder is hypertriglyceridemia with or without a history of pancreatitis.

14. The pharmaceutical composition according to any one of claims 1 to 9, wherein the APOC3-related disease or disorder is familial chylomicronemia syndrome (FCS).

15. The pharmaceutical composition according to any one of claims 1 to 14, wherein the pharmaceutical composition is packaged in a kit, container, pack, dispenser, pre-filled syringe or vial.

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein one or more doses of the pharmaceutical composition are administered by the subject.

17. The pharmaceutical composition according to any one of claims 1 to 16, wherein one or more doses of the pharmaceutical composition are administered by a medical professional.

18. The pharmaceutical composition according to any one of claims 1 to 17, wherein the serum APOC3 protein level is reduced by more than 60% from the baseline level in the human subject.

19. The pharmaceutical composition according to any one of claims 1 to 18, wherein serum triglyceride (TG) levels are reduced by more than 50% from baseline levels in the human subject.

20. The pharmaceutical composition according to any one of claims 1 to 19, wherein serum 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 are reduced in the human subject compared to baseline levels.

21. The pharmaceutical composition according to any one of claims 1 to 20, wherein the subject has a fasting or postprandial baseline triglyceride level higher than 500 mg / dL.

22. The pharmaceutical composition according to any one of claims 1 to 21, wherein the APOC3 RNAi drug substance is administered as a pharmaceutically acceptable salt, a pharmaceutically acceptable mixed salt, a free acid, or a combination thereof.

23. The pharmaceutical composition according to claim 22, wherein the APOC3 RNAi drug substance is administered as a pharmaceutically acceptable sodium salt.

24. A unit dosage form for treating an APOC3-related disease or disorder, wherein the unit dosage form comprises an amount between about 1 mg and about 100 mg of an APOC3 RNAi drug substance, preferably in a pharmaceutically acceptable salt form, wherein the APOC3 RNAi drug substance comprises an antisense strand having the nucleotide sequence: usCfsasCfuGfagaauAfcUfgUfcCfcGfsu (SEQ ID NO: 2) and a sense strand having the nucleotide sequence: (NAG37)s(invAb)sacgggacaGfUfAfuucucaguias(invAb) (SEQ ID NO: 6), where a represents 2'-O-methyladenosine, c represents 2'-O-methylcytidine, and g represents 2'-O-methylguanosine, i represents 2'-O-methylinosine, u represents 2'-O-methyluridine; Af represents 2'-fluoroadenosine, Cf represents 2'-fluorocytidine, Gf represents 2'-fluoroguanosine, Uf represents 2'-fluorouridine; s represents phosphorothioate linkage; (invAb) represents inverted debasic deoxyribose residue; (NAG37)s is 【Chemistry 13】 A unit dosage form containing the structure shown by [the specified method].

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