Methods for preventing cardiovascular events through proprotein convertase subtilisin kexin 9 (PCSK9) protein reduction

Abstract

To provide an injection device for subcutaneous administration, for preventing cardiovascular events in a human subject.SOLUTION: The present invention provides an injection device for subcutaneous administration, comprising a fixed dose of 275 mg to 325 mg of an interfering ribonucleic acid (RNAi) agent. The device comprises a double-stranded ribonucleic acid comprising a sense strand and an antisense strand forming a double-stranded region, where the antisense strand comprises the nucleotide sequence of 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO: 3), and the sense strand comprises the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO: 4), where a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C, or U; Af, Gf, Cf or Uf are 2'-fluoro A, G, C or U; dT is 2'-deoxythymidine; and s is a phosphorothioate linkage.SELECTED DRAWING: None

Description

[Technical field]

[0001] Related Applications This application is a joint venture of U.S. Provisional Patent Application No. 62 / 409,816, filed October 18, 2016; No. 62 / 422,028, filed on March 16, 2017; No. 62 / 472,525; and the benefit of U.S. Provisional Patent Application No. 62 / 550,426, filed August 25, 2017. No. 6,399,623, the entire contents of each of which are incorporated herein by reference.

[0002] This application is also related to U.S. Provisional Patent Application No. 62 / 209,526, filed Aug. 25, 2015; PCT Application No. PCT / US2016 / 048666, filed Dec. 5, 2012; U.S. Provisional Application No. PCT / US2016 / 048666, filed Dec. 5, 2012 No. 61 / 733,518; U.S. Provisional Application No. 61 / 793,530, filed March 15, 2013; U.S. Provisional Application No. 61 / 793,530, filed October 4, 2013 No. 61 / 886,916, filed on Oct. 17, 2013; U.S. Provisional Application No. 61 / 892, filed on Oct. 17, 2013; No. 188; PCT Application No. PCT / US2013 / 073349, filed December 5, 2013; and PCT Application No. PCT / US2013 / 073349, filed June 5, 2015. The provisional patent application and each of the PCT applications are also related to the filed U.S. patent application Ser. No. 14 / 650,128. the entire contents of which are incorporated herein by reference.

[0003] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy, created on October 18, 2017, is incorporated herein by reference in its entirety. It is called 2_3_SL.txt and is 1,799 bytes in size. [Background technology]

[0004] Atherosclerotic cardiovascular disease (ASCVD) remains a global health challenge. It is a systemic condition characterized by the accumulation of fatty deposits, inflammatory cells, and scar tissue within the arterial walls. Atherosclerosis, a complex disease process, is the underlying cause of most clinical cardiovascular events. Various types of treatment and preventive measures have been developed, but low-density lipoprotein Lowering low-protein cholesterol (LDL-C) remains the go-to treatment for combating ASCVD. To this end, statins have been used to lower lipid levels. Although it has become a widely used treatment, the number of subjects at risk is increasing The patient continues to have a normal level of LDL-C.

[0005] Proprotein convertase subtilisin kexin 9 PCSK9 has recently been identified as a target for reducing LDL-C levels. A member of the butyricin serine protease family, it plays a role in cholesterol metabolism. It has been shown to play a role in

[0006] Monoclonal antibodies that block PCSK9 have been developed to reduce circulating PCSK9 levels. However, PCSK9 monoclonal antibodies have not been shown to be effective. However, the duration of treatment is short, which can result in significant administrative and economic burden.

[0007] Thus, reducing LDL-C levels and ultimately treating or preventing ASCVD. There is a need in the art for effective treatment or preventative measures aimed at preventing and treating [Prior art documents] [Patent documents]

[0008] [Patent Document 1] U.S. Provisional Patent Application No. 62 / 209,526 [Patent Document 2] PCT Application No. PCT / US2016 / 048666 [Patent Document 3] U.S. Provisional Application No. 61 / 733,518 [Patent Document 4] U.S. Provisional Application No. 61 / 793,530 [Patent Document 5] U.S. Provisional Application No. 61 / 886,916 [Patent Document 6] U.S. Provisional Application No. 61 / 892,188 [Patent Document 7] PCT Application No. PCT / US2013 / 073349 [Patent Document 8] U.S. Patent Application Serial No. 14 / 650,128 Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention relates to the use of PCSK9 in prophylactic or therapeutic methods for a subject in need thereof. The present invention relates to the use of RNA interference (RNAi) agents to inhibit the formation of [Means for solving the problem]

[0010] In one aspect, the present invention relates to a method of lowering LDL-C in a subject. comprising administering to a subject an effective amount of an RNAi agent, wherein the RNAi agent forms a double-stranded region. The antisense strand is a double-stranded ribonucleic acid having a sense strand and an antisense strand which correspond to SEQ ID NO: The sense strand comprises the nucleotide sequence of SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:4.

[0011] In another aspect, the invention relates to a method of preventing a cardiovascular event in a subject. The method includes administering to a subject an effective amount of an RNAi agent, where the RNAi agent is a double-stranded domain. It is a double-stranded ribonucleic acid that includes a sense strand and an antisense strand that form a region. comprises the nucleotide sequence of SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:4 .

[0012] In yet another aspect, the present invention provides a method for reducing cardiovascular mortality and / or morbidity in a subject. The method includes administering to a subject an effective amount of an RNAi agent, wherein An RNAi agent is a double-stranded ribonucleic acid that includes a sense strand and an antisense strand that form a double-stranded region. The antisense strand contains the nucleotide sequence of SEQ ID NO:3, and the sense strand contains the nucleotide sequence of SEQ ID NO:2. It contains a leutide sequence.

[0013] In certain embodiments, the subject is diagnosed with ASCVD, ASCVD risk equivalent uivalent), increased risk of cardiovascular disease (CVD), heterozygous familial hypercholesterolemia or homozygous familial hypercholesterolemia, and may have low-density lipoprotein If you need to lower your cholesterol, or if not, lower your LDL-C have a disorder, or a combination of disorders, that would benefit from

[0014] In a further aspect, the present invention relates to a method for preventing the onset of ASCVD, the method comprising: administering to a subject an amount of an RNAi agent, wherein the RNAi agent comprises a sensor that forms a double-stranded region, The double-stranded ribonucleic acid comprises a sense strand and an antisense strand. The antisense strand is the nucleic acid of SEQ ID NO: 3. The sense strand comprises the nucleotide sequence of SEQ ID NO:4.

[0015] In another aspect, the present invention provides a method for treating ASCVD, conditions at risk for ASCVD, conditions at increased risk for CVD, Elevated cholesterol, Heterozygous familial hypercholesterolemia, Homozygous familial hypercholesterolemia 2. Treating a subject having, or in need of lowering LDL-C, or a combination thereof. The method includes administering to a subject an effective amount of an RNAi agent, wherein An RNAi agent is a double-stranded ribonucleic acid that includes a sense strand and an antisense strand that form a double-stranded region. The antisense strand contains the nucleotide sequence of SEQ ID NO:3, and the sense strand contains the nucleotide sequence of SEQ ID NO:4. It contains a leutide sequence.

[0016] In certain embodiments, the method includes treating acute coronary syndrome and / or high LDL-C levels. This is for the purpose of

[0017] In some embodiments, the subject has a baseline blood glucose level greater than about 70 mg / dl, e.g., about 100 mg / dl. They may have low LDL-C.

[0018] In certain embodiments, a subject is administered the RNAi agent more than once. The agent may be administered multiple times. In some embodiments, the subject may be administered, for example, about once a week. Once every 2 weeks, once a month, once every 2 months, once every 3 months, once every 4 months, once every 6 months The RNAi agent is administered at regular intervals, such as once a day or approximately once a year.

[0019] In certain embodiments, the methods include evaluating the subject prior to administration of the RNAi agent. measuring one or more biochemical parameters of a subject, including lipid parameters. Examples of parameters that may be measured prior to administration of an RNAi agent include LDL-C, high density lipoprotein HDL-C, PCSK9, total cholesterol, triglycerides, non-HDL-C, ultra Low-density lipoprotein cholesterol (VLDL-C), apolipoprotein A1 (Apo-A1), apolipoprotein A2 (ApoA3), ApoB, lipoprotein(a) (Lp(a)), C-reactive protein (CRP), glycated hemoglobin (GHE) Globin A1c, alanine aminotransferase, aspartate aminotransferase enzymes, alkaline aminotransferase, creatine kinase, and total bilirubin In some embodiments, evaluation of the subject prior to administration of the RNAi agent includes biochemical parameters. In certain embodiments, administration of an RNAi agent provides a baseline measurement of a patient's blood glucose level. The previous subject evaluation may include any appropriate measures regarding administration of the RNAi agent, such as the amount of RNAi agent, the timing of administration of the RNAi agent, etc. Determine and / or influence.

[0020] In certain embodiments, the method includes evaluating the subject between one or more doses of the RNAi agent. The evaluation includes measuring one or more biochemical parameters of the subject, including lipid parameters. The measurement may include measuring the amount of the RNAi agent administered. Examples of parameters are LDL-C, HDL-C, PCSK9, total cholesterol, triglycerides, non-HDL -C, VLDL-C, Apo-A1, Apo-B, Lp(a), CRP, glycosylated hemoglobin A1c, alanine aminotransferase Aspartate aminotransferase, alkaline aminotransferase In some embodiments, the levels of creatine kinase, creatine kinase, and total bilirubin. In this study, the results of the evaluation were used to determine the subsequent administration of the RNAi agent, such as the amount of the RNAi agent and the timing of administration of the RNAi agent. The administration (single or multiple) may be determined and / or influenced.

[0021] In some embodiments, administration of an RNAi agent reduces LDL-C levels relative to baseline LDL-C levels. In certain embodiments, the baseline LDL-C level is reduced by more than about 20%. A reduction in LDL-C levels of more than about 20% compared to baseline levels was observed 15 days after administration of the RNAi agent. or maintained for 15 or more days at a time of 15 or more days.

[0022] In some embodiments, administration of the RNAi agent increases PCSK9 expression levels relative to baseline levels. In certain embodiments, the level of PCSK9 is reduced by more than about 25% compared to baseline. A reduction in the level of PCSK9 of greater than about 25% compared to the level observed within 30 days after administration of the RNAi agent. maintained for 30 or more days during the period above, at a time of 30 or more days.

[0023] In some embodiments, the subject is receiving background lipid-lowering therapy, such as a statin. In certain embodiments, the subject may continue to receive background lipid-lowering therapy. In certain embodiments, the subject is administered an RNAi agent while undergoing treatment with a maximally tolerated The subject may be undergoing statin therapy. Alternatively, the subject may be undergoing ezetimibe or LDL apheresis. may be receiving

[0024] In other embodiments, the subject is not receiving background lipid-lowering therapy.

[0025] In certain embodiments, the subject does not have active liver disease. Alanine aminotransferase and / or aspartate >2x upper limit of normal (ULN) May not provide baseline aminotransferase levels. In the present study, subjects were required to have baseline levels of total bilirubin greater than 1.5 times the ULN. There are times like that.

[0026] In another aspect, the present invention relates to an RNAi agent for use in the following method: (ii) reducing LDL-C in a subject; (iii) preventing a cardiovascular event in a subject; (iii) reducing cardiovascular mortality and / or morbidity in a subject; (iv) reducing ASCV in a subject; (v) to prevent the development of ASCVD, conditions at risk for ASCVD, conditions at increased risk for CVD, and hepatitis; With telozygous familial hypercholesterolemia or homozygous familial hypercholesterolemia The present invention relates to treating a subject having a condition in which the LDL-C level is low, or in need of lowering LDL-C, or a combination thereof. The RNAi agent is a double-stranded ribonucleotide that contains a sense strand and an antisense strand that form a double-stranded region. The antisense strand comprises the nucleotide sequence of SEQ ID NO: 3, and the sense strand comprises the nucleotide sequence of SEQ ID NO: These methods include administering to a subject an effective amount of an RNAi agent. Includes.

[0027] The present invention is further explained by the following detailed description and drawings. [Brief description of the drawings]

[0028] [Figure 1] 1 shows RNAi agents according to embodiments of the present invention. FIG. 1 discloses, in order of appearance, SEQ ID NOs: 4, 3, and 3, respectively. [Diagram 2] 1 shows a summary of the dosing schedules used in the studies discussed in the Examples. [Diagram 3] 1 shows the mean percent change in LDL-C by day 360 following a single administration of an RNAi agent on day 1 as a 200-mg dose, a 300-mg dose, and a 500-mg dose according to an embodiment of the invention. [Figure 4] 1 shows the time-adjusted mean percent change in LDL-C through day 360 following a single administration of an RNAi agent on day 1 as a 200-mg dose, a 300-mg dose, and a 500-mg dose, and following administration of an RNAi agent on days 1 and 90 as a 100-mg dose, a 200-mg dose, and a 300-mg dose, in accordance with an embodiment of the invention. [Diagram 5] 1 shows the time-adjusted mean absolute change in LDL-C through day 360 following a single administration of an RNAi agent on day 1 as a 200-mg dose, a 300-mg dose, and a 500-mg dose, and following administration of an RNAi agent on days 1 and 90 as a 100-mg dose, a 200-mg dose, and a 300-mg dose, according to an embodiment of the invention. [Figure 6A] 1 shows the percent change in LDL-C between baseline and day 270 for individual patients following a single administration of an RNAi agent on day 1 as a 200-mg dose, a 300-mg dose, and a 500-mg dose in accordance with an embodiment of the invention. [Figure 6B] 1 shows the percent change in LDL-C between baseline and day 360 for individual patients following a single administration of an RNAi agent on day 1 as a 200-mg dose, a 300-mg dose, and a 500-mg dose in accordance with an embodiment of the invention. [Figure 7] 1 shows the mean percent change in LDL-C through day 360 following administration of an RNAi agent at days 1 and 90 as a 100-mg dose, a 200-mg dose, and a 300-mg dose according to an embodiment of the invention. [Figure 8] 1 shows the percent change in LDL-C between baseline and day 180 for individual patients following administration of an RNAi agent as a 300-mg dose on days 1 and 90, according to an embodiment of the invention. [Figure 9A] 1 shows the percent change in LDL-C between baseline and day 270 for individual patients following administration of an RNAi agent at days 1 and 90 as a 100-mg dose, a 200-mg dose, and a 300-mg dose in accordance with an embodiment of the invention. [Figure 9B] 1 shows the percent change in LDL-C between baseline and day 360 for individual patients following administration of an RNAi agent at days 1 and 90 as a 100-mg dose, a 200-mg dose, and a 300-mg dose in accordance with an embodiment of the invention. [Figure 10A] 10A-10C show the change in LDL-C from baseline to day 180 for each patient (61 patients) randomly assigned to the two-dose placebo group according to an embodiment of the invention. The dashed lines represent LDL-C reductions of 39 mg per deciliter and 78 mg per deciliter (multiply by 0.02586 to convert cholesterol values ​​to millimoles per liter). [Figure 10B] FIG. 1 shows the change in LDL-C from baseline to day 180 for each patient (59 patients) randomly assigned to the two-dose 300-mg RNAi agent group according to an embodiment of the invention. The dashed lines represent LDL-C reductions of 39 mg per deciliter and 78 mg per deciliter (multiply by 0.02586 to convert cholesterol values ​​to millimoles per liter). [Figure 10C] FIG. 1 shows the change in LDL-C from baseline to day 240 for each patient (59 patients) randomly assigned to the two-dose 300-mg RNAi agent group according to an embodiment of the invention. The dashed lines represent LDL-C reductions of 39 mg per deciliter and 78 mg per deciliter (multiply by 0.02586 to convert cholesterol values ​​to millimoles per liter). [Figure 11A]1 shows the percent change in LDL-C from baseline to days 90 and 270 for individual patients following administration of an RNAi agent on day 1 as a 300-mg dose according to an embodiment of the invention. [Figure 11B] 1 shows the percent change in LDL-C from baseline to days 90 and 270 for individual patients following administration of an RNAi agent as a 300 mg dose on days 1 and 90 according to an embodiment of the invention. [Figure 12] 1 shows the mean percent change in PCSK9 levels by day 270 following a single administration of an RNAi agent on day 1 as a 200-mg dose, a 300-mg dose, and a 500-mg dose according to an embodiment of the invention. [Figure 13] 1 shows the mean percent change in PCSK9 levels by day 270 following administration of an RNAi agent at days 1 and 90 as a 100-mg dose, a 200-mg dose, and a 300-mg dose according to an embodiment of the invention. [Figure 14] 1 shows modeled results for LDL-C up to 22 months following administration of an RNAi agent administered as a 300-mg dose twice or three times a year, according to an embodiment of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] The present invention relates, at least in part, to a method for reducing levels of PCSK9 protein. It is based on the use of small interfering RNA (siRNA) molecules, which then lower LDL-C levels. Through RNA interference, siRNA binds to the RNA-induced silencing complex (RISC) in cells. This allows it to cleave the messenger RNA (mRNA) that codes for PCSK9. The truncated mRNA is degraded and therefore unavailable for protein translation, resulting in the transcription of PSCK9. Protein levels are reduced, and as a result, LDL-C levels are reduced.

[0030] Accordingly, the present invention provides a method for lowering LDL-C in a subject, comprising administering a prophylactically effective amount or The present invention provides a method comprising administering to a subject an effective, such as a therapeutically effective, amount of an RNAi agent. The disclosure also provides a method of preventing a cardiovascular event in a subject, comprising administering a prophylactically effective amount or a therapeutically effective amount of The present invention further provides a method comprising administering to a subject an effective amount of an RNAi agent, such as an amount A method of reducing cardiovascular mortality and / or morbidity in a subject, comprising administering to a subject a prophylactically effective amount of or a therapeutically effective amount of an RNAi agent. For these methods, subjects were classified as having ASCVD, ASCVD and risk equivalent, or at increased risk of CVD. , heterozygous familial hypercholesterolemia, and homozygous familial hypercholesterolemia have a need to lower LDL-C, or a combination thereof.

[0031] The present invention further provides a method for preventing the onset of ASCVD in a subject, comprising administering to a subject a prophylactically effective amount or The present invention provides a method comprising administering to a subject an effective, such as a therapeutically effective, amount of an RNAi agent.

[0032] In addition, the present invention relates to a method for treating ASCVD, conditions at risk for ASCVD, conditions at risk for CVD, and heterogeneous Having homozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, Methods of Treating a Subject Having a Need for Lowering LDL-C, or Combinations Thereof The method includes administering to a subject an effective amount, such as a prophylactically effective amount or a therapeutically effective amount, of an RNAi agent. The method may include a step of:

[0033] The present invention further provides a method for the treatment of a subject comprising administering to said subject a therapeutic agent ... for the treatment of a subject. to lower LDL-C in subjects; to prevent cardiovascular events in subjects; reducing cardiovascular mortality and / or morbidity in subjects; preventing the onset of ASCVD in subjects; and / or ASCVD, ASCVD-related risk conditions, elevated risk of CVD, hepatitis, Patients with bizygous familial hypercholesterolemia or homozygous familial hypercholesterolemia The present invention relates to treating a subject having a need to lower LDL-C, and RNAi agents for use in

[0034] In some embodiments, the cardiovascular event is death, non-fatal myocardial infarction, severe relapse, or Major adverse events include, but are not limited to, acute ischemia, stroke, symptomatic pulmonary embolism, and hemorrhage. Adverse cardiovascular events may occur.

[0035] The methods of lowering LDL-C and / or preventing a cardiovascular event include treating a patient with ASCVD or or to subjects who are at "risk for ASCVD." In certain embodiments, In this study, subjects were classified as having "a condition equivalent to ASCVD risk" if they had one or more of the following: ": Cardiac risk assessed by Framingham risk score (score > 20%) or equivalent method Symptomatic Attenuated Vascular Disease (HAVD) In some embodiments, the present invention relates to atherosclerosis, type 2 diabetes, and familial hypercholesterolemia. Alternatively, the subject may have heterozygous familial hypercholesterolemia.

[0036] The methods of lowering LDL-C and / or preventing cardiovascular events also include treating ASCVD. are at "risk for ASCVD" or otherwise develop heart health problems The present invention may be directed to a subject who exhibits one or more symptoms or risk factors that may be associated with the development of a disease. Such symptoms / risk factors include having an acute coronary syndrome; myocardial infarction; stable or unstable angina. coronary or other arterial revascularization, stroke, transient ischemic attack, or atheroma history of peripheral arterial disease of atherosclerotic origin; male sex; had a family history of conditions associated with ASCVD risk; smoked tobacco; and were physically inactive. not being of legal age; having high blood pressure; having high blood cholesterol; having diabetes or pre-diabetes having a medical condition; being overweight or obese; having a history of preeclampsia during pregnancy and; inability to manage stress and / or anger; being post-menopausal; and , an unhealthy diet, such as a diet high in trans fats, cholesterol, and / or refined sugars; Being 55 years of age or older; having sleep apnea; being anemic; or any combination thereof Thus, the method may involve, for example, matching a subject who is young but has a family history of heart disease. In certain embodiments, the method may be directed to subjects who smoke but do not have high blood pressure. may be directed to subjects who have previously experienced a cardiac event.

[0037] In some embodiments, the methods of the invention involve administering an effective amount, such as a prophylactically effective amount or a therapeutically effective amount. administering an RNAi agent to a subject having heterozygous familial hypercholesterolemia. For lowering LDL-C and / or preventing cardiovascular events in In some embodiments, the methods of the present invention include administering a prophylactically or therapeutically effective amount. and administering an effective amount of an RNAi agent to a subject having ASCVD. and / or for preventing cardiovascular events. In embodiments, the methods of the invention involve administering an effective amount of an RNAi agent, such as a prophylactically effective amount or a therapeutically effective amount. and lowering LDL-C in a subject having a condition at risk for ASCVD, comprising administering to said subject a therapeutically effective amount of LDL-C to said subject. Some of the results may be about preventing cardiovascular events and / or cardiovascular disease. In embodiments, the methods of the invention involve administering an effective amount, such as a prophylactically effective amount or a therapeutically effective amount, of an RNAi agent. The present invention relates to a method for reducing LDL-C in a subject with homozygous familial hypercholesterolemia, the method comprising the steps of: The objective of the present invention may be to reduce cardiovascular disease and / or prevent cardiovascular events. In some embodiments, the methods of the invention involve administering an effective amount of an RNAi A method for treating a subject with heterozygous familial hypercholesterolemia comprising administering to said subject a In some embodiments, the methods of the invention can be about predicting The method includes administering an effective amount, such as a preventively or therapeutically effective amount, of an RNAi agent to a patient having ASCVD. In some embodiments, the methods of the present invention may be for treating a subject. The method includes administering an effective amount, such as a prophylactically or therapeutically effective amount, of an RNAi agent to treat ASCVD and / or ASCVD-associated cancer. The risk may be for treating a subject having a comparable condition. In embodiments, the methods of the invention involve administering an effective amount, such as a prophylactically effective amount or a therapeutically effective amount, of an RNAi agent. The present invention relates to a method for treating a subject having homozygous familial hypercholesterolemia, comprising administering to said subject a method for treating homozygous familial hypercholesterolemia. In these embodiments, the RNAi agent can be a sense gene that forms a double-stranded region. and an antisense strand, wherein the antisense strand is a double-stranded ribonucleic acid having a sequence The subject is a subject's nucleotide sequence comprising SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:4. , fat may be receiving a diet designed to improve their quality of life and / or may be taking statins, You may be taking LDL-lowering therapy such as ezetimibe or LDL apheresis. In some embodiments, the subject may be on maximally tolerated statin therapy. Additionally, the subject may require further reduction in LDL-C.

[0038] In certain embodiments, administration of an RNAi agent reduces LDL-C levels by about 2% compared to baseline LDL-C levels. More than 0%, or more than about 25%, or more than about 30%, or more than about 35%, or more than about 40% or about 45% or more, or about 50% or more, or about 55% or more, or about 60% or more It can also reduce LDL-C levels.

[0039] In some embodiments, the reduction in the level of LDL-C after the RNAi agent is administered is indicative of the RNAi About 15 days or more, about 20 days or more, about 30 days or more, about 40 days or more, or about 50 days after administration of the agent or about 60 days or more, or about 70 days or more, or about 80 days or more, or about 90 days or more, or about 100 days or more days or more, or about 110 days or more, or about 120 days or more, or about 130 days or more, or about 140 days or more, or About 150 days or more, or about 160 days or more, or about 170 days or more, or about 180 days or more, or about 190 days or more or about 200 days or more, or about 210 days or more, or about 220 days or more, or about 230 days or more, or about 240 days or more days or more, or about 250 days or more, or about 260 days or more, or about 270 days or more, or about 280 days or more, or About 290 days or more, or about 300 days or more, or about 310 days or more, or about 320 days or more, or about 330 days or more or for about 340 days or more, or about 350 days or more, or about 360 days or more, at the time of these dates, or may be maintained for these days.

[0040] Administration of RNAi agents at doses of approximately 100 mg reduced baseline LDL-C levels by day 15. In comparison, LDL-C levels may be reduced by more than about 15%, or more than about 20%. The reduction in the amount of β-glucose (β-glucose) is observed for about 30 days, or about 60 days, or about 90 days, or for a period of time longer than these days. In some embodiments, the vein can be maintained at or over these number of days. the reduction in LDL-C compared to baseline LDL-C levels may be greater than about 40% at day 30; For about 60 days, about 90 days, or longer, at or near these dates It can be maintained.

[0041] Administration of RNAi agents at doses of approximately 200 mg reduced baseline LDL-C levels by day 15. In comparison, about 15% more, or about 20% more, or about 25% more, or about 30% more This reduction in LDL-C can occur over about 30 days, or about 60 days, or about 90 days. or about 120 days, or about 150 days, or about 180 days, or about 210 days, or about 240 days, or about 270 days, or for about 300 days, or about 330 days, or about 360 days, or more, The baseline may be maintained at or over these number of days. The reduction in LDL-C compared to baseline LDL-C levels can be greater than about 40% at 30 days, greater than about 90% at about 60 days, and greater than about 100% at about 120 days. 0 days or for a longer period, at or over these days In certain embodiments, the reduction in LDL-C compared to baseline LDL-C levels can be , greater than about 30% at 15 days, about 30 days, about 60 days, about 90 days, about 120 days, about 150 days, or It may be maintained for longer periods of time, at these times, or over these times. In some embodiments, the reduction in LDL-C compared to the baseline LDL-C level is at least 180 days after administration. In certain embodiments, the baseline L The reduction in LDL-C relative to DL-C levels can be greater than about 20% at 15 days, greater than about 30 days, or greater than about 60 days, or 90 days, or about 120 days, or about 150 days, or about 180 days, or about 210 days, or about 240 days or for about 270 days, or for about 300 days, or for about 330 days, or for about 360 days, at the time of these dates, or may be maintained over these number of days. In certain embodiments, the baseline LDL- The reduction in LDL-C relative to C levels is greater than about 25% at 180 days, or greater than about 25% at 270 days, or may be about 30% greater at 360 days.

[0042] Administration of an RNAi agent at a dose of approximately 300 mg resulted in a significant increase in LDL-C levels by day 15 compared to baseline. and increasing the level of LDL-C by more than about 15%, or more than about 20%, or more than about 25%, or more than about 30%, or more than about 40%. This reduction in LDL-C can last for about 30 days, or for about 60 days, or for about 90 days, or for about 120 days, or about 150 days, or about 180 days, or about 210 days, or about 240 days, or about 270 days, or about 300 days, or For about 330 days, or about 360 days, or more, at or about the time of these dates In some embodiments, the LDL-C level is increased relative to baseline LDL-C levels and maintained over a period of days. The reduction in LDL-C compared to the control group can be greater than about 45% at 30 days, and greater than about 60 days, about 90 days, or longer. The method may be continued for a period of time, at a time point, or over a period of time. In this embodiment, the reduction in LDL-C compared to baseline LDL-C levels is about 40% or more, for about 30 days, about 60 days, about 90 days, about 120 days, about 150 days, or more; It may be maintained at or for these number of days. In the study, the reduction in LDL-C compared to baseline LDL-C levels was greater than approximately 35% at day 15. about 30 days, or about 60 days, or about 60 days, or about 90 days, or about 120 days, or about 150 days, or for about 180 days, or about 210 days, or for a longer period of time, or at the time of In some embodiments, the baseline LDL-C level and the The comparative LDL-C reduction may be greater than about 30% at 15 days, greater than about 30 days, greater than about 60 days, or greater than about 90 days, or about 120 days, or about 150 days, or about 180 days, or about 210 days, or about 240 days, or about 270 days days, or for about 300 days, or for about 330 days, or for about 360 days, or for a longer period of time, The baseline may be established at any time point or over a period of these days. The reduction in LDL-C compared to baseline LDL-C levels was greater than about 35% at 180 days or greater than about 35% at 270 days. or greater than about 30% at 360 days.

[0043] Administration of an RNAi agent at a dose of approximately 500 mg resulted in a significant increase in LDL-C levels by day 15 compared to baseline. and increasing the level of LDL-C by more than about 15%, or more than about 20%, or more than about 25%, or more than about 30%, or more than about 40%. This reduction in LDL-C can last for about 30 days, or for about 60 days, or for about 90 days, or for about 120 days, or about 150 days, or about 180 days, or about 210 days, or about 240 days, or about 270 days, or about 300 days, or For about 330 days, or about 360 days, or more, at or about the time of these dates In some embodiments, the LDL-C level is increased relative to baseline LDL-C levels and maintained over a period of days. The relative LDL-C reduction can be greater than about 50% at 30 days, and greater than about 60 days, about 90 days, or longer. The method may be continued for a period of time, at a time point, or over a period of time. In this embodiment, the reduction in LDL-C compared to baseline LDL-C levels is about It may be more than 40%, about 30 days, about 60 days, about 90 days, about 120 days, about 150 days, about 180 days, or more. The method may be continued for a number of days, at a time point of a number of days, or over a number of days. In embodiments, the reduction in LDL-C compared to baseline LDL-C levels is about 35% or more at day 15. %, and may be about 30 days, or about 60 days, or about 90 days, or about 120 days, or about 150 days, or about 180 days. 210 days, or about 240 days, or for a period of time longer than these days, In some embodiments, the reduction in LDL-C can be maintained over these days. may be greater than about 30% at day 15, greater than about 30 days, or greater than about 60 days, compared to baseline LDL-C levels; or about 90 days, or about 120 days, or about 150 days, or about 180 days, or about 210 days, or about 240 days, or for about 270 days, or about 300 days, or for a period of time greater than or equal to these dates. In certain embodiments, the LDL-C level is increased relative to baseline LDL-C levels and can be maintained over a period of days. The reduction in LDL-C compared to the control group was greater than about 40% at 180 days, greater than about 30% at 270 days, or greater than about 30% at 360 days. On the day, the concentration may be greater than about 30%.

[0044] Administration of a second dose of the RNAi agent on day 90 reduced LDL-C levels compared to baseline LDL-C levels. The level of LC can be further reduced. For example, administration of a dose of about 100 mg of an RNAi agent on day 1 After administration of a dose of about 100 mg of the RNAi agent at 90 days, compared to baseline LDL-C levels , >40% between days 104 and 120 and / or 150; >35% by day 104 and / or day 180; >35% by day 104 to day 120, day 150, day 180 and / or day >30% by day 210; Days 104 to 120, 150, 180, 210, and 240 and / or greater than about 25% by day 270; days 104 to 120, 150, 180, and 210 , greater than about 20% by day 240, day 270, day 300, and / or day 330; and / or by day 104 From the 120th day, 150th day, 180th day, 210th day, 240th day, 270th day, 300th day, 330th day and and / or by 360 days, the LDL-C level may be reduced by more than about 10%. Compared to baseline LDL-C levels, the reduction in LDL-C was greater than about 35% at 180 days, and may be greater than about 25% at 270 days, or greater than about 10% at 360 days.

[0045] Administration of about a 200 mg dose of the RNAi agent on day 1, followed by administration of about a 200 mg dose of the RNAi agent on day 90 Treatment was continued from days 104 to 120 and / or 150 compared to baseline LDL-C levels. >45% by day 104 to day 120, day 150, day 180, day 210, and / or day 240 >40% by day 104; Days 104 to 120, 150, 180, 210, 240, and 270 and / or greater than about 35% by day 300; and / or greater than about 35% by day 104 to day 120, day 150, day 180 Approximately 30% by the 210th, 240th, 270th, 300th, 330th, and / or 360th day In certain embodiments, the LDL-C level can be reduced by more than 100%. In comparison, the reduction in LDL-C was greater than about 40% at 180 days, or greater than about 35% at 270 days, may be greater than about 30% at 360 days.

[0046] Administration of about a 300 mg dose of the RNAi agent on day 1, followed by administration of about a 300 mg dose of the RNAi agent on day 90 Treatment was compared to baseline LDL-C levels on days 104 to 120, 150, and 180. and / or greater than about 50% by day 210; days 104 to 120, 150, 180, and 210 , >40% by Day 240, Day 270, and / or Day 300; Days 104 to 120, Day 150 , greater than about 35% by day 180, day 210, day 240, day 270, day 300, and / or day 310; and / or days 104 to 120, 150, 180, 210, 240, 270, 30 The levels of LDL-C may be reduced by more than about 30% by day 0, day 330, and / or day 360. In certain embodiments, the reduction in LDL-C compared to the baseline LDL-C level is at least 180 days after administration. or greater than about 50% at day 270, or greater than about 40% at day 270, or greater than about 30% at day 360. do.

[0047] In certain embodiments, administration of the RNAi agent reduces PCSK9 levels, as compared to baseline: More than about 25%, or more than about 30%, or more than about 35%, or more than about 40%, or more than about 45% %, or about 50% or more, or about 55% or more, or about 60% or more, or about 65% or more or about 50% or more, or about 70% or more, or about 75% or more, or about 80% or more or by about 85% or more, or by about 90% or more, or by about 95% or more. It can be done.

[0048] In some embodiments, the reduction in the level of PCSK9 is greater than or equal to about 15 days after administration of the RNAi agent, or about 20 days or more, or about 30 days or more, or about 40 days or more, or about 50 days or more, or about 60 days or more, or about 70 days or more, or about 80 days or more, or about 90 days or more, or about 100 days or more, or about 110 days or more, or about 120 days or more, or about 130 days or more, or about 140 days or more, or about 150 days or more, or about 160 days or about 170 days or more, or about 180 days or more, or about 190 days or more, or about 200 days or more, or about 210 days or more, or about 220 days or more, or about 230 days or more, or about 240 days or more, or about 250 days or more, or about 260 days or more, or about 270 days or more, or about 280 days or more, or about 290 days or more, or about 300 days or about 310 days or more, or about 320 days or more, or about 330 days or more, or about 340 days or more, or about At or over a period of 350 days or more, or approximately 360 days or more It can be maintained.

[0049] Administration of an RNAi agent at a dose of approximately 100 mg resulted in a significant improvement in PCSK9 levels compared to baseline by day 15. and about 20% or more, or about 25% or more, or about 35% or more, or about 40% or more of PC The reduction in PCSK9 may be achieved for about 30 days, or for about 60 days, or for about 90 days. or for a longer period of time, at or over these days obtain.

[0050] Administration of the RNAi agent at a dose of approximately 200 mg resulted in a significant improvement in PCSK9 levels compared to baseline by day 15. and about 20% or more, or about 25% or more, or about 30% or more, or about 35% or more, or may reduce the level of PCSK9 by more than about 40%, or more than about 45%, or more than about 50%. This reduction in PCSK9 may last for about 30 days, or for about 60 days, or for about 90 days, or for longer. In some embodiments, the PCSK9 level is increased relative to baseline. In this case, the reduction in PCSK9 may be greater than about 60% at 30 days, about 60 days, about 90 days, or longer. The present invention may be applied to a particular patient, a particular patient group, or a particular patient group. In embodiments, the reduction in PCSK9 compared to baseline PCSK9 levels is about 3 to about 300 days. It may be greater than 0%.

[0051] Administration of the RNAi agent at a dose of approximately 300 mg resulted in a significant improvement in PCSK9 levels compared to baseline by day 15. and about 20% or more, or about 25% or more, or about 30% or more, or about 35% or more, or is greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about The reduction in PCSK9 may be achieved over a period of about 30 days or over a period of about 60 days. The method may be maintained for up to about 90 days, or for up to about 90 days, or for longer. Compared to baseline PCSK9 levels, the reduction in PCSK9 was greater than approximately 60% at 30 days. Possibly, for about 60 days, about 90 days, or longer, at or about the time of these dates. In certain embodiments, the baseline PCSK9 level may be maintained for a period of time ranging from 1 to 4 days. In comparison, the reduction in PCSK9 can be greater than about 30% by day 300.

[0052] Administration of a dose of approximately 500 mg of the RNAi agent resulted in a significant increase in PCSK9 levels compared to baseline by day 15. and about 20% or more, or about 25% or more, or about 30% or more, or about 35% or more, or is greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about The reduction in PCSK9 may be achieved over a period of about 30 days or over a period of about 60 days. The method may be maintained for up to about 90 days, or for up to about 90 days, or for longer. Compared to baseline PCSK9 levels, the reduction in PCSK9 was greater than approximately 70% at 30 days. It may be that the drug is administered for about 60 days, about 90 days, or longer, at or after these dates. In some embodiments, the baseline PCSK9 level and the In comparison, reduction in PCSK9 can be greater than about 30% by day 300.

[0053] Administration of about a 100 mg dose of the RNAi agent on day 1 followed by about a 100 mg dose of the RNAi agent on day 90 Administration of 10 mg / kg / day increased PCSK9 levels by approximately 20% or more compared to baseline by day 104. or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, PCSK9 This reduction in PCSK9 may occur for about 120 days, at about 120 days, or at about 1 In some embodiments, the PCSK9 level is increased relative to the baseline PCSK9 level and maintained for 20 days. In comparison, the reduction in PCSK9 may be greater than about 30% at day 104 and for about 150 days. The effect may be maintained for approximately 150 days or for up to about 150 days.

[0054] Administration of an approximately 200 mg dose of the RNAi agent on day 1 followed by an approximately 200 mg dose of the RNAi agent on day 90 Administration of 10 mg / kg / day increased PCSK9 levels by approximately 20% or more compared to baseline by day 104. is about 25% or more, or about 30% or more, or about 35% or more, or about 40% or more, or about The levels of PCSK9 may be reduced by more than 45%, or by more than about 50%. The method may be continued for about 120 days, at about 120 days, or over about 120 days. In terms of morphology, the reduction in PCSK9 was approximately 4% at day 104 compared to baseline PCSK9 levels. 0% and may be maintained for, at, or over about 150 days.

[0055] Administration of an approximately 300 mg dose of the RNAi agent on day 1 followed by an approximately 300 mg dose of the RNAi agent on day 90 Administration of 10 mg / kg / day increased PCSK9 levels by approximately 20% or more compared to baseline by day 104. is about 25% or more, or about 30% or more, or about 35% or more, or about 40% or more, or about The levels of PCSK9 may be reduced by more than 45%, or by more than about 50%. Maintained for about 120 days, at about 120 days, or throughout about 120 days, or throughout 150 days In some embodiments, a decrease in PCSK9 as compared to baseline PCSK9 levels may be observed. The reduction may be greater than about 50% or greater than about 55% at 120 days, for about 150 days, at about 150 days, Or it may be maintained for about 150 days.

[0056] Administration of RNAi agents at doses of approximately 100 mg reduced baseline total cholesterol by day 90. Compared to the level, about 5% more, about 10% more, about 15% more, or or by about 20% or more; by day 90, about 5% more, about 10% more, or about 15% more, compared to the non-HDL-C level of or about 20% or more, or about 25% or more, or about 30% or more of non-H By day 90, Apo-B levels were reduced by approximately 5% compared to baseline levels. or about 10% or more, or about 15% or more, or about 20% or more, or by about 25% or more; and / or by day 90, or about 5% or more, or about 10% or more, or about 15% or more, compared to the Lp(a) level in the More specifically, it may reduce the levels of Lp(a).

[0057] Administration of RNAi agents at doses of approximately 200 mg reduced baseline total cholesterol by day 90. Compared to the level, about 5% more, about 10% more, about 15% more, or or by about 20% or more, or by about 25% or more, to reduce total cholesterol levels; By day 1, the non-HDL-C level is increased by about 5% or by about 10% compared to the baseline non-HDL-C level. or about 15% or more, or about 20% or more, or about 25% or more, or by about 30% or more, or by about 35% or more; by day 90 5% or more, or about 10% or more, or slightly higher than baseline Apo-B levels. or about 15% or more, or about 20% or more, or about 25% or more, or about 30% and / or reduce baseline Lp(a) levels by Day 90. about 5% or more, or about 10% or more, or about 15% or more, or In some embodiments, the Lp(a) level may be reduced by more than about 20% by day 180. In the study, total cholesterol was reduced by more than 15%, non-HDL-C by more than 25%, and Apo-B by more than 20%. It's super.

[0058] Administration of RNAi agents at doses of approximately 300 mg reduced baseline total cholesterol by day 90. Compared to the level, about 5% more, about 10% more, about 15% more, or or by about 20% or more, or by about 25% or more, to reduce total cholesterol levels; By day 1, the non-HDL-C level is increased by about 5% or by about 10% compared to the baseline non-HDL-C level. or about 15% or more, or about 20% or more, or about 25% or more, or greater than about 30%, greater than about 35%, or greater than about 40% of the non-HDL-C level. by day 90, Apo-B levels were reduced by approximately 5% compared to baseline levels or about 10% or more, or about 15% or more, or about 20% or more, or about 25% reduce levels of Apo-B by more than about 30%, or by more than about 35%; and / or by Day 90, a decrease in Lp(a) levels by more than about 5% compared to baseline or by increasing the level of Lp(a) by about 10%, about 15%, or about 20%. It can be reduced.

[0059] Administration of RNAi agents at doses of approximately 500 mg reduced baseline total cholesterol by day 90. Compared to the level, about 5% more, about 10% more, about 15% more, or or about 20% or more, about 25% or more, or about 30% or more of the total cholesterol by day 90, it reduced HDL-C levels by approximately 5% compared to baseline non-HDL-C levels or about 10% or more, or about 15% or more, or about 20% or more, or about 25% or more, or about 30% or more, or about 35% or more, or about 40% by 90 days compared to baseline Apo-B levels In comparison, about 5% more, or about 10% more, or about 15% more, or about 20% more %, or about 25% or about 30% or about 35% or more; or by more than about 40%; and / or by day 90, about 5% or about 10% or about 15% greater than the normal Lp(a) level In some embodiments, the Lp(a) level may be reduced by more than about 20% or by more than about 20%. At 180 days, total cholesterol was reduced by more than 25% and Apo-B by more than 30%. do.

[0060] Administration of about a 100 mg dose of the RNAi agent on day 1 followed by about a 100 mg dose of the RNAi agent on day 90 Administration of 100 mg ... or about 10% or more, or about 15% or more, or about 20% or more, Reduced cholesterol levels; by day 180, compared to baseline non-HDL-C levels and about 5% or more, or about 10% or more, or about 15% or more, or about 20% or by about 25% or more, or by about 30% or more. By day 180, there was an increase of about 5% or about 1% compared to baseline Apo-B levels. More than 0%, or more than about 15%, or more than about 20%, or more than about 25% and / or reduce baseline Lp(a) levels by day 180. In comparison, the level of Lp(a) may be reduced by greater than about 5%, or greater than about 10%.

[0061] Administration of an approximately 200 mg dose of the RNAi agent on day 1 followed by an approximately 200 mg dose of the RNAi agent on day 90 Administration of 100 mg ... or about 10% or more, or about 15% or more, or about 20% or more, or by about 25% or more; by day 180, about 5% or more, about 10% or more, or about 15% or more compared to the non-HDL-C level of the patient or about 20% or more, or about 25% or more, or about 30% or more, or reduce non-HDL-C levels by about 35% or more; by Day 180, reduce baseline Ap oB level, about 5% or more, about 10% or more, or about 15% or more or about 20% or more, or about 25% or more, or about 30% or more, or reduce Apo-B levels by more than about 35%; and / or reduce baseline L p(a) levels, or , may reduce levels of Lp(a).

[0062] Administration of an approximately 300 mg dose of the RNAi agent on day 1 followed by an approximately 300 mg dose of the RNAi agent on day 90 Administration of 100 mg ... or about 10% or more, or about 15% or more, or about 20% or more, or by about 25% or more, or by about 30% or more, to reduce total cholesterol levels; By day 1, the non-HDL-C level is increased by about 5% or by about 10% compared to the baseline non-HDL-C level. or about 15% or more, or about 20% or more, or about 25% or more, or about 30% or more, about 35% or more, about 40% or more, about 45% by day 180, compared to baseline Apo-B levels In comparison, about 5% more, or about 10% more, or about 15% more, or about 20% more %, or about 25% or about 30% or about 35% or more; or by more than about 40%; and / or by day 180, about 5% or about 10% or about 15% higher than the normal Lp(a) level %, or by about 20% or by about 25% or more. obtain.

[0063] In certain embodiments, the method further comprises evaluating the subject prior to administration of the RNAi agent. Assessment may be performed on the same day, if prior to administration of the RNAi agent, or may be performed one day prior, or two days prior, if prior to administration of the RNAi agent. Days before, or 3 days before, or 5 days before, or 6 days before, or 7 days before, or 8 days before, or 9 days before, or 10 days before It may be performed 11, 12, 13, or 14 days before. In embodiments, the assessment may be performed two or more days prior to administration of the RNAi agent.

[0064] In some embodiments, the assessment includes determining age, height, weight, body mass index, race, sex, Whether the subject was receiving any other treatment (e.g., lipid-lowering therapy such as statins), cardiac Including but not limited to vascular disease diagnosis and diabetes diagnosis, heart rate, blood pressure, ECG parameters, etc. may include measuring one or more physiological parameters or characteristics of the subject without do.

[0065] In certain embodiments, the evaluation comprises measuring one or more biochemical parameters of the subject. In some embodiments, the value measured before administration of the RNAi agent may be referred to as a "baseline" value. Examples of biochemical parameters, including lipid parameters, are , LDL-C, HDL-C, PCSK9, total cholesterol, triglycerides, non-HDL-C, VLDL-C, Apo-A1 , Apo-B, Lp(a), CRP, glycosylated hemoglobin A1c, alanine aminotransferase, As Paraglutamine aminotransferase, alkaline aminotransferase, creatine The levels of erythrocyte kinase, total bilirubin, and erythrocyte stimuli may be, but are not limited to, those listed above. In one embodiment, evaluation of the subject prior to administration of the RNAi agent includes baseline biochemical parameters. In certain embodiments, evaluation of the subject prior to administration of the RNAi agent may provide a measure of the responsiveness of the subject to the RNAi agent. Deciding and / or administering the RNAi agent, e.g., the amount of RNAi agent, the timing of administration of the RNAi agent, etc. can have an impact.

[0066] In certain embodiments, the method comprises evaluating the subject between one or more doses of the RNAi agent. The evaluation can include evaluating the expression level during administration, e.g., 1 day, 2 days, 3 days, 4 days, or after administration of the RNAi agent. , 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 20th, 21st, 28th , 30th, 35th, 40th, 42nd, 45th, 49th, 50th, 56th, 60th, 63rd, 70th, 77th, 80th, One or more biochemical parameters of the subjects, including lipid parameters, are evaluated during the 84 and / or 90 days. The biochemical measurements may include measuring LDL-C, HDL-C, PCSK9, Total cholesterol, triglycerides, non-HDL-C, VLDL-C, Apo-A1, apolipoprotein B, Lp(a), CRP, glycosylated hemoglobin A1c, alanine aminotransferase, asparagine Acid aminotransferase, alkaline aminotransferase, creatine kinase , and total bilirubin levels.

[0067] In some embodiments, the assessment during one or more doses of the RNAi agent includes assessing body weight, fat mass, and the like. Biomedical parameters include, but are not limited to, blood pressure index, heart rate, blood pressure, electrocardiogram parameters, etc. A physical parameter or characteristic of the subject may be measured.

[0068] In some embodiments, the results of the assessment determine the amount of RNAi agent, the timing of administration of the RNAi agent, Such may determine and / or affect subsequent administration(s) of an RNAi agent, such as For example, one or more results from the measurements may indicate a subsequent reduction in the dosage of the RNAi agent by about 5%, or about 10%, or less. or about 20%, or about 30%, or about 40%, or about 50%, or about 60%, or may be increased or decreased by about 70%, or about 80%, or about 90%, or more.

[0069] Various biochemical parameters may determine / influence the dose of RNAi agent administered. For example, alanine aminotransferase or aspartate aminotransferase >2x the ULN Measurements of transferase and / or total bilirubin greater than 1.5 times the ULN should be followed by RNA analysis. As another example, a measurement of glycated hemoglobin A1c of more than 10% may result in a reduction in the dosage of anti-inflammatory drugs. The set point may result in a subsequent reduction in the dose of the RNAi agent. A reduction in LDL-C measurements of less than about 5%, less than about 10%, or less than about 15% is considered to be a significant improvement over the subsequent R This may result in an increase in the dose of the NAi agent administered. The dose may be, for example, about 1%, or about 5%, or about 10%. , or about 15%, or about 20%, or about 25%, or about 30%, or about 35%, or about 40%, or about 45%, Or it may be decreased or increased by about 50%, or more.

[0070] In certain embodiments, after the subject is assessed prior to administration of the RNAi agent, as described above, the subject is The subject may be administered lipid-lowering therapy. Lipid-lowering therapy is a treatment in which the subject is administered lipid-lowering therapy on a regular basis. In some embodiments, the subject may be a patient with a prior history of lipid-lowering therapy. In certain embodiments, after administration of a lipid-lowering treatment regimen, The subject is evaluated for one or more biochemical parameters of the subject, including lipid parameters, as described above. The subject may then undergo additional evaluation to measure the level of oxidative stress and / or oxidative stress. of RNAi agent may be administered.

[0071] Administration of RNAi agents The methods of the invention include the steps of administering to a subject an effective, such as a prophylactically or therapeutically effective, amount of an RNAi agent. Includes.

[0072] A "prophylactically effective amount" refers to an amount that is sufficient to treat an individual at risk of contracting a condition, but who has not yet experienced or exhibited symptoms of the condition. Prevents or ameliorates a condition or one or more symptoms of the condition when administered to a subject having the condition. Ameliorating a condition can include slowing the progression of a condition. A "prophylactically effective amount" includes a therapeutically effective amount of an RNAi agent, Method of administration of the drug, risk of the condition, and medical history, age, weight, family history, genetic makeup, if any May vary depending on type of prior or concomitant therapy, as well as other individual characteristics of the patient being treated. .

[0073] A "therapeutically effective amount" refers to an amount that, when administered to a subject, is sufficient to treat (e.g., an existing condition or one or more sufficient to effect treatment of a condition (by alleviating, ameliorating, or maintaining the symptoms of a number of conditions) A "therapeutically effective amount" can include an amount of an RNAi agent that is sufficient to treat a patient with a disease that is at least partially or completely treatable with the agent. and medical history, age, weight, family history, genetic makeup, stage of pathological process, if any, antecedent or concomitant The duration of treatment may vary depending on the type of treatment being used, and other individual characteristics of the patient being treated.

[0074] "Baseline" may refer to a treatment-free state, such as before treatment is administered.

[0075] The RNAi agent may be administered to the subject as a fixed dose. A "fixed dose" (e.g., a dose in mg) is The dose used for all subjects, regardless of any particular subject-related factors such as body weight Alternatively, the RNAi agent is administered to the subject as a weight-based dose (e.g., a dose in mg / kg). The subject may be administered multiple doses of the RNAi agent, with the dose varying depending on the subject's weight. In embodiments where a dose is administered, the RNAi agent is administered in a combination of a fixed dose and a weight-based dose. The drug may be administered in a single dose.

[0076] In certain embodiments, the RNAi agent is about 50 mg to about 800 mg, about 100 mg to about 800 mg, about 150 mg to about 500 mg, about 200 mg to about 500 mg, about 250 mg to about 500 mg, about 300 mg to about 600 mg, about 400 mg to about 700 mg, about 500 mg to about 800 mg, about 600 mg to mg to about 800 mg, about 200 mg to about 800 mg, 250 mg to about 800 mg, about 300 mg to about 800 mg, about 350 mg to about 800 mg, about 400 mg to about 800 mg, about 450 mg to about 800 mg, about 500 mg to about 800 mg, about 550 mg to about 800 mg, about 600 mg to about 800 mg, about 650 mg to about 800 mg, about 700 mg to about 800 mg, about 750 mg to about 800 mg, about 50 mg to about 750 mg, about 100 mg to about 750 mg, about 150 mg to about 750 mg, about 200 mg to about 750 mg, about 250 mg to about 750 mg, about 300 mg to about 750 mg, about 350 mg to about 750 mg, about 400 mg to about 750 mg, about 450 mg to about 750 mg, about 500 mg to about 750 mg, about 550 mg to about 750 mg, about 600 mg to about 750 mg, about 650 mg to about 750 mg, about 700 mg to about 750 mg, about 50 mg to about 700 mg, about 100 mg to about 700 mg, about 150 mg to about 700 mg, about 200 mg to about 700 mg, about 250 mg to about 700 mg, about 300 mg to about 700 mg, about 350 mg to about 700 mg, about 400 mg to about 700 mg, about 450 mg to about 700 mg, about 500 mg to about 700 mg, about 550 mg to about 700 mg, about 600 mg to about 700 mg, about 650 mg to about 700 mg, about 50 mg to about 650 mg, about 100 mg to about 650 mg, about 150 mg to about 650 mg, about 200 mg to about 650 mg, about 250 mg to about 650 mg, about 300 mg to about 650 mg, about 350 mg to about 650 mg, about 400 mg to about 650 mg, about 450 mg to about 650 mg, about 500 mg to about 650 mg, about 550 mg to about 650 mg, about 600 mg to about 650 mg, about 50 mg to about 600 mg, about 100 mg to about 600 mg, about 150 mg to about 600 mg, about 200 mg to about 600 mg, about 250 mg to about 600 mg, about 300 mg to about 600 mg, about 350 mg to about 600 mg, about 400 mg to about 600 mg, about 450 mg to about 600 mg, about 500 mg to about 600 mg, about 550 mg to about 600 mg, about 50 mg to about 550 mg, about 100 mg to about 550 mg, about 150 mg to about 550 mg, about 200 mg to about 550 mg, about 250 mg to about 550 mg, about 300 mg to about 550 mg, about 350 mg to about 550 mg, about 400 mg to about 550 mg, about 450 mg to about 550 mg, about 500 mg to about 550 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 150 mg to about 500 mg, about 200 mg to about 500 mg, about 250 mg to about 500 mg, about 300 mg to about 500 mg, about 350 mg to about 500 mg, about 400 mg to about 500 mg, about 450 mg to about 500 mg, about 50 mg to about 450 mg, about 100 mg to about 450 mg, about 150 mg to about 450 mg, about 200 mg to about 450 mg, about 250 mg to about 450 mg, about 300 mg to about 450 mg, about 350 mg to about 450 mg, about 400 mg to about 450 mg, about 50 mg to about 400 mg, about 100 mg to about 400 mg, about 150 mg to about 400 mg, about 200 mg to about 400 mg, about 250 mg to about 400 mg, about 300 mg to about 400 mg, about 350 mg to about 400 mg, about 50 mg to about 350 mg, about 100 mg to about 350 mg, about 150 mg to about 350 mg, about 200 mg to about 350 mg, about 250 mg to about 350 mg, about 300 mg to about 350 mg, about 50 mg to about 300 mg, about 100 mg from about 150 mg to about 300 mg, from about 200 mg to about 300 mg, or from about 250 mg to about 300 mg of solid Fixed doses, for example, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 2 25mg, about 250mg, about 275mg, about 300mg, about 325mg, about 350mg, about 375mg, about 400mg, about 425mg, about 450mg, about 475mg, about 500mg, about 525mg, about 550mg, about 575mg, about 600mg, about 625mg, about 650mg, Administered to a subject as a fixed dose of about 675 mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg, or about 800 mg. Values ​​and ranges intermediate to those recited above are also intended to be part of the invention. will be done.

[0077] The RNAi agent can be administered, for example, in multiple doses repeated at regular intervals. Ai agents are available for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, and about 4 days. Weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 1 year, about 13 months, about 14 months, about 15 months, about 16 months 1 month, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months The compound may be administered to a subject at intervals of a month or more, e.g., chronic administration. So, fixed doses can be administered once or multiple times a year, i.e., 2, 3, 4, 5, 6, 7, 8, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd The subject may be administered 1, 23, 24 or more times. The amount is approximately every 4 weeks, approximately every 5 weeks, approximately every 6 weeks, approximately every 7 weeks, approximately every 8 weeks, approximately every 9 weeks, approximately every 10 weeks every 11 weeks, every 12 weeks, every 13 weeks, every 14 weeks, every 15 weeks, every 16 weeks, every 17 weeks Every week, about every 18 weeks, about every 19 weeks, about every 20 weeks, about every 22 weeks, about every 24 weeks, about every 26 weeks, Every 28 weeks, Every 30 weeks, Every 32 weeks, Every 34 weeks, Every 36 weeks, Every 38 weeks, Every 40 weeks every about 42 weeks, about every 44 weeks, about every 46 weeks, about every 48 weeks, about every 50 weeks, about every 52 weeks, or The subject may be administered once per day for a longer period of time. In certain embodiments, the fixed dose is Once per day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once every 7 days, once every 8 days Once a day, once every 9 days, once every 10 days, once every 11 days, once every 12 days, once every 13 days, once every 14 days Once every 15 days, once every 16 days, once every 17 days, once every 18 days, once every 19 days, once every 20 days Once per 30 days, once per 40 days, once per 50 days, once per 60 days, once per 70 days, once per 80 days once every 90 days, once every 100 days, once every 110 days, once every 120 days, once every 130 days, once every 140 days once every 150 days, once every 160 days, once every 170 days, once every 180 days, once every 190 days, once every 2 Once every 00 days, once about 210 days, once about 220 days, once about 230 days, once about 240 days, once about 250 days , once every 260 days, once every 270 days, once every 280 days, once every 290 days, once every 300 days, once every 310 days once every 320 days, once every 330 days, once every 340 days, once every 350 days, once every 360 days, once every 3 The subject may be administered once every 65 days or longer.

[0078] In certain embodiments, the RNAi agent is administered in a "loading phase" of closely spaced administrations followed by and a dosing regimen that includes a "maintenance phase" in which the RNAi agent may be administered at longer intervals. For example, administration may be once a week or once every two weeks for one month, followed by administration once a month, This may be repeated for six months or a year or longer (eg, chronic administration).

[0079] In some embodiments, the loading phase includes the first week, the first two weeks, the first three weeks This may include administration of an RNAi agent for the first month, etc.

[0080] In certain embodiments, one or more doses may be administered during the loading phase. The loading phase includes an initial administration of the RNAi agent on day 1, followed by administration of the RNAi agent on about days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, About 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 30 days, about 40 days, about 50 days, about 60 days, About 70 days, about 80 days, about 90 days, about 100 days, about 110 days, about 120 days, about 130 days, about 140 days, about 150 days, about 160 days, about 170 days, about 180 days, about 190 days, about 200 days, about 210 days, about 220 days, about 230 days, about 240 days, Approximately 250 days, 260 days, 270 days, 280 days, 290 days, 300 days, 310 days, 320 days, 330 days, One or more Rs after about 340 days, about 350 days, about 360 days, about 365 days, or more In some embodiments, the loading phase may include administration of an NAi agent on day 1. and then administering the RNAi agent for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, 8 weeks, 9 weeks, 10 weeks, 13 weeks, 15 weeks, 20 weeks, 25 weeks, 26 weeks weeks, about 30 weeks, about 35 weeks, about 40 weeks, about 45 weeks, about 50 weeks, about 52 weeks, or longer and administering a second dose of the RNAi agent shortly thereafter. The phases consisted of administration of the RNAi agent on day 1, followed by administration for about 1 month, 2 months, 3 months, and 4 months. months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, or longer after a second administration of the RNAi agent. In this case, the loading phase may include additional doses of the RNAi agent (i.e., more than two doses), Thus, the interval between each administration may vary.

[0081] In certain embodiments, the RNA agent may be administered on day 1 and again on or around day 90. .

[0082] In certain embodiments, the RNAi agent is administered at a dose of about 50 mg to about 800 mg, about 100 mg to about 500 mg, or about 100 mg to about 500 mg during the loading phase. about 150 mg to about 800 mg, about 200 mg to about 800 mg, 250 mg to about 800 mg, about 300 mg to about 800 mg About 800mg, about 350mg to about 800mg, about 400mg to about 800mg, about 450mg to about 800mg, about 500mg to About 800mg, about 550mg, about 800mg, about 600mg to about 800mg, about 650mg to about 800mg, about 700mg to about 800mg, about 750mg to about 800mg, about 50mg to about 750mg, about 100mg to about 750mg, about 150mg to about 7 50mg, about 200mg to about 750mg, about 250mg to about 750mg, about 300mg to about 750mg, about 350mg to about 7 50mg, about 400mg to about 750mg, about 450mg to about 750mg, about 500mg to about 750mg, about 550mg to about 7 50mg, about 600mg to about 750mg, about 650mg to about 750mg, about 700mg to about 750mg, about 50mg to about 70 0 mg, about 100 mg to about 700 mg, about 150 mg to about 700 mg, about 200 mg to about 700 mg, about 250 mg to about 70 0 mg, about 300 mg to about 700 mg, about 350 mg to about 700 mg, about 400 mg to about 700 mg, about 450 mg to about 70 0 mg, about 500 mg to about 700 mg, about 550 mg to about 700 mg, about 600 mg to about 700 mg, about 650 mg to about 70 0 mg, about 50 mg to about 650 mg, about 100 mg to about 650 mg, about 150 mg to about 650 mg, about 200 mg to about 650 mg, about 250 mg to about 650 mg, about 300 mg to about 650 mg, about 350 mg to about 650 mg, about 400 mg to about 650 mg mg, about 450mg to about 650mg, about 500mg to about 650mg, about 550mg to about 650mg, about 600mg to about 650mg mg, about 50 mg to about 600 mg, about 100 mg to about 600 mg, about 150 mg to about 600 mg, about 200 mg to about 600 mg g, about 250 mg to about 600 mg, about 300 mg to about 600 mg, about 350 mg to about 600 mg, about 400 mg to about 600 mg g, about 450 mg to about 600 mg, about 500 mg to about 600 mg, about 550 mg to about 600 mg, about 50 mg to about 550 mg , about 100 mg to about 550 mg, about 150 mg to about 550 mg, about 200 mg to about 550 mg, about 250 mg to about 550 mg , about 300 mg to about 550 mg, about 350 mg to about 550 mg, about 400 mg to about 550 mg, about 450 mg to about 550 mg , about 500 mg to about 550 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 150 mg to about 500 mg , about 200 mg to about 500 mg, about 250 mg to about 500 mg, about 300 mg to about 500 mg, about 350 mg to about 500 mg , about 400 mg to about 500 mg, about 450 mg to about 500 mg, about 50 mg to about 450 mg, about 100 mg to about 450 mg , about 150 mg to about 450 mg, about 200 mg to about 450 mg, about 250 mg to about 450 mg, about 300 mg to about 450 mg , about 350 mg to about 450 mg, about 400 mg to about 450 mg, about 50 mg to about 400 mg, about 100 mg to about 400 mg , about 150 mg to about 400 mg, about 200 mg to about 400 mg, about 250 mg to about 400 mg, about 300 mg to about 400 mg , about 350 mg to about 400 mg, about 50 mg to about 350 mg, about 100 mg to about 350 mg, about 150 mg to about 350 mg , about 200 mg to about 350 mg, about 250 mg to about 350 mg, about 300 mg to about 350 mg, about 50 mg to about 300 mg , about 100 mg to about 300 mg, about 150 mg to about 300 mg, about 200 mg to about 300 mg, or about 250 mg to about 3 As a fixed dose of about 100 mg, for example, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg , about 200mg, about 225mg, about 250mg, about 275mg, about 300mg, about 325mg, about 350mg, about 375mg, about 400m g, about 425 mg, about 450 mg, about 475 mg, about 500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about 625 mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg, or about 800 mg fixed dose can be administered to a subject. Values ​​and ranges intermediate to those listed above are also part of the invention. It is intended that

[0083] In some embodiments, each dose of an RNAi agent administered during the loading phase is administered at the same dose. The amounts may be the same or different.

[0084] In certain embodiments, the RNAi agent is administered as a dose of about 300 mg on day 1 and at about 90 days. It may be administered as a dose of about 300 mg around or on the 90th day.

[0085] In certain embodiments, the maintenance phase comprises administering to the subject one or more doses of an RNAi agent. Administration may include once a month, once every two months, once every three months, once every four months, once every five months, once every six months, once every 7 months, once every 8 months, once every 9 months, once every 10 months, once every 11 months, once every 12 months , once a year, once every 13 months, once every 14 months, once every 15 months, once every 16 months, once every 17 months, once every 18 months Once a month, once a month 19, once a month 20, once a month 21, once a month 22, once a month 23, once a month 24 Once per month, once per 25 months, once per 26 months, once per 27 months, once per 28 months, once per 29 months, once per 30 months once every 31 months, once every 32 months, once every 33 months, once every 34 months, once every 35 months, once every 36 months , once every 37 months, once every 38 months, once every 39 months, once every 40 months, once every 41 months, once every 42 months, once every 43 once every month, once every 44 months, once every 45 months, once every 46 months, once every 47 months, once every 48 months, or In one particular embodiment, the maintenance dose is once every three months or once every six months. It is administered once a month, once every nine months, or once a year.

[0086] In an embodiment of the present invention, the maintenance phase may be, for example, a multiple use phase repeated at regular intervals. For example, the RNAi agent can be administered for about 1 day, about 2 days, about 3 days, about 4 days, About 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months , about 1 year, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 19 months, At intervals of about 20 months, about 21 months, about 22 months, about 23 months, about 24 months, or longer, e.g. For example, the fixed dose may be administered to a subject chronically. In certain embodiments, the fixed dose may be administered once or multiple times a year. Several times, i.e., 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, or more frequently In some embodiments, the fixed dose is administered to the subject about every 4 weeks, about every 5 weeks, about every 10 weeks, about every 15 weeks, about every 20 weeks, about every 25 weeks, about every 30 weeks, about every 40 weeks, about every 5 ... Every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about Every 13 weeks, about every 14 weeks, about every 15 weeks, about every 16 weeks, about every 17 weeks, about every 18 weeks, about every 19 weeks , approximately every 20 weeks, approximately every 22 weeks, approximately every 24 weeks, approximately every 26 weeks, approximately every 28 weeks, approximately every 30 weeks, approximately every 32 weeks every 34 weeks, every 36 weeks, every 38 weeks, every 40 weeks, every 42 weeks, every 44 weeks, every Administered to a subject once every 46 weeks, about every 48 weeks, about every 50 weeks, about every 52 weeks, or longer. In certain embodiments, the fixed dose can be administered once a day, about once every two days, about once every three days, about once every four days, about once every five days, about once every six days, about once every seven days, about once every eight ... Once every 4 days, once every 5 days, once every 6 days, once every 7 days, once every 8 days, once every 9 days, once every 10 days , once every 11 days, once every 12 days, once every 13 days, once every 14 days, once every 15 days, once every 16 days , once every 17 days, once every 18 days, once every 19 days, once every 20 days, once every 30 days, once every 40 days, Once every 50 days, once every 60 days, once every 70 days, once every 80 days, once every 90 days, once every 100 days, Once every 110 days, once every 120 days, once every 130 days, once every 140 days, once every 150 days, once every 160 days 170 days, 180 days, 190 days, 200 days, 210 days, 220 days Once a day, once every 230 days, once every 240 days, once every 250 days, once every 260 days, once every 270 days, Once every 280 days, once every 290 days, once every 300 days, once every 310 days, once every 320 days, once every 330 days once every 340 days, once every 350 days, once every 360 days, once every 365 days or longer, It may be administered to elephants.

[0087] In certain embodiments, the maintenance phase may be administered about every 3 months, about every 4 months, about every 6 months, or about every 9 months. This can include administering a dose of an RNAi agent to a subject monthly, or about once a year.

[0088] The maintenance dose(s) may be the same as or different from the loading dose(s). For example, The maintenance dose administered to a subject is from about 25 mg to about 800 mg, for example, about 25 mg, about 50 mg, about 75 mg, About 100mg, about 125mg, about 150mg, about 175mg, about 200mg, 225mg, about 250mg, about 275mg, about 300mg, About 325mg, about 350mg, about 375mg, about 400mg, 425mg, about 450mg, about 475mg, about 500mg, about 525mg, About 550mg, about 575mg, about 600mg, about 625mg, about 650mg, about 675mg, about 700mg, 725mg, about 750mg, It may be about 775 mg, or about 800 mg. Intermediate values ​​and ranges among those recited above are also within the scope of the present invention. It is intended to be part of.

[0089] In certain embodiments, the maintenance phase is every 3 months, every 4 months, every 6 months, or every 9 months. or annually to a subject.

[0090] In certain embodiments, the RNAi agent is administered in a dose of: (1) 300 mg on day 1 and on about day 90; and (2) a loading phase in which the RNAi agent is administered at 300 mg approximately every 6 months after administration on or around day 90. The drug may be administered in a dosing regimen that includes a maintenance phase in which the drug is administered.

[0091] RNAi agents can be administered subcutaneously, intravenously, intramuscularly, intraocularly, intrabronchially, intrapleurally, intraperitoneally, intraarterially, or intralymphatically. Intracerebral, intracerebrospinal, and any combination thereof, The composition may be administered to a subject using any mode of administration known in the art. The drug is administered subcutaneously.

[0092] In some embodiments, administration is via depot injection. Depot injections can be subcutaneous or intramuscular. In certain embodiments, the depot injection is a subcutaneous injection.

[0093] In some embodiments, administration is via a pump, which may be an external pump or a surgically implanted pump. In certain embodiments, the pump can be a subcutaneously implanted percutaneous pump. In another embodiment, the pump is an infusion pump. The pump is used for intravenous, subcutaneous, intra-arterial, or epidural infusion. In certain embodiments, the infusion pump may be a subcutaneous infusion pump. In another embodiment, the pump is a surgically implanted pump that delivers RNAi agents to the liver. It is a pump that has been designed.

[0094] Other modes of administration include epidural, intracerebral, intraventricular, intranasal, intraarterial, intracardiac, intraosseous, intrathecal, intrathecal. Modes of administration include intrathecal, intravitreal, pulmonary, oral, topical, intratracheal, epidermal, or transdermal. Based on whether local or systemic treatment is desired and on the area to be treated The route and site of administration may be selected to enhance targeting.

[0095] RNAi agents were administered at approximately 1 min, 2 min, 3 min, 4 min, 5 min, 6 min, 7 min, 8 min, 9 min, 10 min, 11 min, 12 min, and 13 min. minutes, 14 minutes, 15 minutes, 16 minutes, 17 minutes, 18 minutes, 19 minutes, 20 minutes, 21 minutes, 22 minutes, 23 minutes, 24 minutes, 25 minutes, 26 minutes , 27 minutes, 28 minutes, 29 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, or 60 minutes or more Administration can be by intravenous infusion for a period of time such as every week, every other week, or the like. (i.e. every 2 weeks), etc., periodically for 1 month, 2 months, 3 months, 4 months, or longer After the initial treatment regimen, treatment may be administered less frequently. For example, after administration once a week or once every two weeks for three months, administration may be discontinued for six months or one year. Or for longer, it may be repeated once a month.

[0096] The RNAi agent may be administered in combination with one or more other prophylactic or therapeutic agents. The agent may be in the same formulation as one or more other prophylactic or therapeutic agents, or may be administered in combination with a RNase inhibitor. The Ai agent may be in a different formulation from one or more other prophylactic or therapeutic agents. The agent and one or more other prophylactic or therapeutic agents may be administered simultaneously or within a short period of each other, e.g. For example, within about 1 minute, within about 5 minutes, within about 15 minutes, within about 30 minutes, within about 60 minutes, or Within about 2 hours, or within about 3 hours, or within about 4 hours, or within about 6 hours, or within about 9 hours or within about 12 hours, or within about 15 hours, or within about 18 hours, or within about 24 hours It can be done.

[0097] The one or more other prophylactic or therapeutic agents may be hypercholesterolemia, atherosclerosis, The present invention also includes those known to treat lipid disorders such as diabetes, obesity, and dyslipidemia. For example, the one or more other prophylactic or therapeutic agents may be an HMG-CoA reductase inhibitor, a fibrinogen activator, Glucocorticoids, bile acid sequestrants, niacin, antiplatelet agents, angiotensin-converting enzyme inhibitors, angiotensin Diotensin II receptor antagonist, acyl-CoA cholesterol acetyltransferase (AC AT) inhibitors, cholesterol absorption inhibitors, cholesterol ester transfer protein CETP inhibitors, microsomal triglyceride transfer protein (MTTP) inhibitors , cholesterol regulators, bile acid regulators, peroxisome proliferator-activated receptor (PPAR) receptors , gene-based therapies, and combination vasoprotectants (e.g., AGI-1067 from Atherogenics) , glycoprotein IIb / IIIa inhibitors, aspirin or aspirin-like compounds, IBAT inhibitors, The inhibitor may be an allene synthase inhibitor, or a monocyte chemotactic protein (MCP)-I inhibitor. As an example, the one or more other prophylactic or therapeutic agents may be, for example, alirocumab (Pralent). (Repata), evolocumab (Repata), bococizumab, roderucizumab, ralpanizumab, RG7 The antibody may be an anti-PCSK9 antibody, such as 652, LY3015014, LPD1462, AX1, ALD1306, or Ig1-PA4.

[0098] Subjects receiving RNAi agents The subject to which the RNAi agent is administered may be a human or a non-human animal, and in certain embodiments, It may be a vertebrate, and in particular embodiments a mammal. The elephant is a human. In certain embodiments, the subject is an adult. In some embodiments, , the subjects are patients.

[0099] The subject may have a baseline LDL-C level of about 70 mg / dl or greater. In the present invention, the baseline LDL-C level is about 80 mg / dl or more, or about 90 mg / dl or more, or about 100 mg / dl or more. / dl or more, or about 110 mg / dl or more, or about 120 mg / dl or more, or about 130 mg / dl or more, or about 140 mg / dl or more, or about 150 mg / dl or more, or about 160 mg / dl or more, or about 170 mg / dl or more. / dl or more, or about 190 mg / dl or more, or about 200 mg / dl or more, or about 210 mg / dl or more, or about 220 mg / dl or more, or about 230 mg / dl or more, or about 240 mg / dl or more, or about 250 mg / dl or more, or about 260 mg / dl or more, or about 270 mg / dl or more, or about 280 mg / dl or more, or about 290 mg / dl or more, or about 300 mg / dl or higher.

[0100] In some embodiments, the subject is in need of lowering LDL-C. The subject has a blood glucose level of about 50 mg / dl or more, or about 60 mg / dl or more, or about 70 mg / dl or more, or about 80 mg / dl or more. or about 90 mg / dl or more, or about 100 mg / dl or more, or about 110 mg / dl or more, or about 120 mg / dl or more or about 130 mg / dl or more, or about 140 mg / dl or more, or about 150 mg / dl or more, or about 160 mg / dl or more or about 170 mg / dl or more, or about 180 mg / dl or more, or about 190 mg / dl or more, or about 200 mg / dl or more or about 210 mg / dl or more, or about 220 mg / dl or more, or about 230 mg / dl or more, or about 240 mg / dl or more or about 250 mg / dl or more, or about 260 mg / dl or more, or about 270 mg / dl or more, or about 280 mg / dl or more or greater than about 290 mg / dl, or greater than about 300 mg / dl.

[0101] In some embodiments, the subject does not have active liver disease. Aminotransferase, aspartate aminotransferase, and total bilirubin The biochemical parameters of the IL-1 gene may be determined by measuring one or more of the following biochemical parameters: Biochemical parameters may be measured at baseline. In certain embodiments, The subject may have an alanine aminotransferase level less than or equal to two times (2x) the ULN. In some embodiments, the subject has an ALK level of about 1.5×ULN, or about the ULN, or below the ULN. The subject may have an aminotransferase level.

[0102] In some embodiments, the subject has an aspartate aminotransferase level less than or equal to two times (2×) the ULN. In certain embodiments, the subject may have a transaminase level of about 1.5×ULN, or have aspartate aminotransferase levels at or below the ULN obtain.

[0103] In some embodiments, the subject may have a total bilirubin level of 1.5×ULN or less. In certain embodiments, the subject has a total bilirubin level at about or below the ULN. possible.

[0104] Alternatively, the subject may have active liver disease. In certain embodiments, the subject is About 2.5 x ULN, or about 3 x ULN, or about 3.5 x ULN, or about 4 x ULN, or more, twice the ULN In some embodiments, the alanine aminotransferase level is greater than 2×. In some embodiments, the subject has a serum IL-10 level of about 2.5×ULN, or about 3×ULN, or about 3.5×ULN, or about 4×ULN, or therebetween. have aspartate aminotransferase levels greater than 2x the ULN, i.e., supra In certain embodiments, the subject receives a serum IL-10 level of about 2×ULN, about 2.5×ULN, or about 3×ULN, or about Have a total bilirubin level greater than 1.5 x ULN, such as 3.5 x ULN, or about 4 x ULN, or greater possible.

[0105] Alanine aminotransferase, aspartate aminotransferase, and The ULN for total bilirubin will be understood by one of skill in the art.

[0106] In some embodiments, the subject may be undergoing background lipid-lowering therapy. Subjects may continue to receive background lipid-lowering therapy while receiving an RNAi agent. Alternatively, the subject may discontinue background lipid-lowering therapy. In the present study, the background lipid-lowering therapy may be a statin, an example of which is atorvastatin. statin, pravastatin, simvastatin, lovastatin, fluvastatin, cerivastatin These include, but are not limited to, statin, rosuvastatin, and pitavastatin. Patients may be receiving maximally tolerated statin therapy; the maximum dose of statin is determined by the art. It is well known in the art.

[0107] In some embodiments, the subject is receiving treatment with ezetimibe, LDL apheresis, bile acid sequestrants, Other types of lipid-lowering therapy may be used, such as nicotinic acid and fibrates.

[0108] In other embodiments, the subject is not receiving background lipid-lowering therapy.

[0109] In certain embodiments, the subject is receiving a diet designed to improve, for example, lipid levels. Such diets are known in the art and may be administered in a controlled manner. For example, diets designed to improve lipid levels may be associated with eating less red meat. and; removing fat from meat; not eating fried foods or high-fat sauces; not eating egg yolks. Skim milk or 1% milk, low-fat frozen yogurt, low-fat ice cream and low-fat dairy products, such as low-fat cheese; and low-fat sources of fiber, such as fruits and vegetables. This may include eating foods that are

[0110] The subject may have a triglyceride level of about 400 mg / dl or less. g / dl, or about 360 mg / dl, or about 340 mg / dl, or about 320 mg / dl, or about 300 mg / dl, or about 280 mg g / dl, or about 260 mg / dl, or about 240 mg / dl, or about 220 mg / dl, or about 200 mg / dl, or about 180 mg g / dl, or about 160 mg / dl, or about 140 mg / dl, or about 120 mg / dl, or about 100 mg / dl, or about 80 mg / dl, or about 60 mg / dl.

[0111] In other embodiments, the subject may have a triglyceride level greater than about 400 mg / dl. , the subject has a blood glucose level of about 420 mg / dl, or about 440 mg / dl, or about 460 mg / dl, or about 480 mg / dl, or about 500 mg / dl. g / dl, or about 520 mg / dl, or about 540 mg / dl, or about 560 mg / dl, or about 580 mg / dl, or about 600 mg The subjects may have a baseline triglyceride level of g / dl.

[0112] In some embodiments, triglyceride levels are measured at baseline.

[0113] The subject may have an estimated glomerular filtration rate (eGFR) of at least about 30 ml / min. For example, the eGFR is , about 30 ml / min, or about 35 ml / min, or about 40 ml / min, or about 45 ml / min, or about 50 ml / min, or It can be more than that.

[0114] In some embodiments, the subject has hyperlipidemia, such as hypercholesterolemia. In certain embodiments, the subject has heterozygous familial hypercholesterolemia. In other embodiments, the subject has homozygous familial hypercholesterolemia. .

[0115] Alternatively, the subject may have an eGFR of less than about 30 ml / min, such as about 25 ml / min, or about 20 ml / min, or about 15 ml / min. may have an eGFR of less than 0.05 mg / kg.

[0116] In some embodiments, the eGFR is measured at baseline.

[0117] The subject may have untreated or inadequately or not at all addressed diabetes mellitus, such as type 2 diabetes. , and may not have poorly controlled type 2 diabetes. Diabetes can be identified by a glycated hemoglobin A1c level of at least about 10%. Thus, the subject has less than about 10%, for example, about 9%, or about 8%, or about 7%, or about 6%, or about 5%, or may have a glycated hemoglobin A1c level below that.

[0118] In other embodiments, the subject may have poorly controlled type 2 diabetes, e.g. For example, at least about 10% of the glycated hemoglobin is greater than about 15%, or about 20%, or about 25%, or about 30%. This can be evidenced by Robin A1c levels.

[0119] In some embodiments, the glycated hemoglobin A1c level is measured at baseline.

[0120] Subjects were characterized by the New York Heart Association (NYHA) as class II, III, or IV. Alternatively, the subject may not have heart failure as defined by the New York Heart Association (NYHA). The patient may have heart failure that is better characterized as Class II, III, or IV.

[0121] The subject may have 30% or more ventricles, such as about 35%, or about 40%, or about 45%, or about 50%, or more. In other embodiments, the subject may have an ejection fraction of about 25%, or about 20%, or about 15%, or less. The patient may have a ventricular ejection fraction of less than 30%, such as a ventricular ejection fraction of less than 100%. The ventricular ejection fraction may be measured as a measure of the ventricular ejection fraction (VEF) in the ventricular ejection chamber, or may be the last known ventricular ejection fraction previously measured. This is also possible.

[0122] The subject may not experience a major adverse cardiac event within 6 months of administration of the RNAi agent. Adverse cardiac events were death, nonfatal myocardial infarction, severe recurrent ischemia, stroke, and symptomatic events. In some embodiments, the primary The risk of adverse cardiac events was highest within 7 months, 8 months, 9 months, and 10 months of administration of the RNAi agent. Within months, within about 11 months, within about 12 months, within about 18 months, within about 24 months, within about 30 months, Within approximately 36 months or longer, including without limitation the occurrence of a major cardiac event In alternative embodiments, the subject has not experienced the disease within about 5 months, or within about 4 months. Within a month, or within about three months, or within about two months, or within about one month, or within about four weeks, or Within about 3 weeks, or within about 2 weeks, or within about 1 week, or earlier. Patients may have experienced a major cardiac event within 6 months of receiving the drug.

[0123] The subject may not have severe hypertension. The blood pressure is about 190 mmHg, or about 200 mmHg, or about 220 mmHg, or about 240 mmHg, or about 260 mmHg. or about 280 mmHg, or about 300 mmHg, or a systolic blood pressure of more than about 180 mmHg. and / or a systolic blood pressure of about 120 mmHg, or about 140 mmHg, or about 160 mmHg, or about 180 mmHg. or a diastolic blood pressure of about 200 mmHg, or about 220 mmHg, or about 240 mmHg, or more. In certain embodiments, the subject may be identified by a diastolic blood pressure greater than about 110 mmHg. Patients may not have severe, uncontrolled hypertension. Hypertension is defined as a systolic blood pressure greater than about 180 mmHg and / or a blood pressure greater than about 11 mmHg despite antihypertensive therapy. The subject may be identified by a diastolic blood pressure greater than 0 mmHg. Alternatively, the subject may be identified by severe hypertension and / or Or they may have severe uncontrolled hypertension.

[0124] In some embodiments, blood pressure is measured at baseline.

[0125] The subject may not have a history of any hemorrhagic stroke. May have had a stroke.

[0126] The subject is within about 4 months, or within about 5 months, or within about 6 months, or within about 7 months, or within about Within 8 months, or within about 9 months, or within about 10 months, or within about 11 months, or within about 12 months and no cardiac arrhythmias within 3 months of administration of the RNAi agent, including within a longer period of time. In other embodiments, the subject has had a cardiac arrhythmia within 3 months of administration of the RNAi agent. Alternatively, the cardiac arrhythmia is managed, for example, by medication or by ablation. The subject has had a cardiac arrhythmia that has not been controlled, e.g., by medication or ablation, within the last three months. It may be possible.

[0127] In certain embodiments, the subject may have one or more of the following: myocardial infarction , stable or unstable angina, coronary or other arterial revascularization, stroke, transient ischemic attack or a history of peripheral arterial disease of atherosclerotic origin; male sex; cardiac Having a family history of ASCVD or conditions associated with risk of ASCVD; being physically inactive; having high blood pressure; high blood cholesterol; diabetes having diabetes and pre-diabetes; being overweight or obese; pre-eclampsia during pregnancy having a history of depression; having unmanageable stress or anger; being postmenopausal; For example, a diet high in salt, saturated fat, trans fat, cholesterol, and / or refined sugar any unhealthy diet; being over 55 years old; having sleep apnea; being anemic; or A combination of them.

[0128] In some embodiments, the subject is diagnosed with, e.g., Alzheimer's disease, dementia, memory loss. In some embodiments, the subject may have any cognitive impairment. It has no harm.

[0129] RNAi agents RNAi agents are double-stranded ribonucleic acids that contain a sense strand and an antisense strand that form a double-stranded region. The antisense strand has the nucleotide sequence of SEQ ID NO: 1, i.e., 5'-ACAAAAGCAAAACAGG The sense strand contains the nucleotide sequence of SEQ ID NO: 2, i.e. 5'-CUAGACCUGU Contains TUUGCUUUUGU-3'.

[0130] In an embodiment of the invention, at least one nucleotide on the antisense strand, At least one nucleotide on the antisense strand, or on both the antisense and sense strands In some embodiments, at least one nucleotide in is a modified nucleotide. , substantially every nucleotide of the antisense strand, substantially every nucleotide of the sense strand or substantially all nucleotides of both the antisense and sense strands are modified nucleotides. In certain embodiments, every nucleotide of the antisense strand, every nucleotide of the sense strand, All nucleotides of the antisense strand or all nucleotides of both the antisense and sense strands are modified nucleotides. It is a nucleotide.

[0131] Modifications to nucleotides are made using techniques and methods known in the art. The modifications are incorporated herein by reference in PCT Application No. PCT / US2016 / 048666, filed August 25, 2016, and in PCT Application No. PCT / US2016 / 048667, filed June 2015. No. 14 / 650,128, filed on the 5th, which is incorporated herein by reference. No. 6,399,433, which is incorporated herein by reference.

[0132] In some embodiments, one or more nucleotides of the antisense or sense strand are The tides may be 2'-O-methyl (2'-OMe) or 2'-fluoro (2'-F) modified. In embodiments, one or more nucleotides of the antisense or sense strand are 3'-5' - may be connected via a phosphodiester linkage.

[0133] In some embodiments, the double-stranded ribonucleic acid comprises a ligand. In this method, the ligand is conjugated to the 3' end of the sense strand of a double-stranded ribonucleic acid. Examples of applications are PCT Application No. PCT / US2016 / 048666, filed August 25, 2016, and PCT Application No. PCT / US2016 / 048667, filed June 5, 2015. No. 14 / 650,128, filed on the same day, which are incorporated herein by reference. In certain embodiments, the ligand is N-acetylgalactosamine. (GalNAc) derivative.

[0134] In certain embodiments, the RNAi agent is 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' ( The antisense strand of the nucleotide sequence of SEQ ID NO: 3) and 5'-csusagacCfuGfudTuugcuuuugu-3' A double-stranded ribonucleic acid comprising a sense strand of the nucleotide sequence of (SEQ ID NO: 4), wherein a, g , c and u are 2'-O-methyl (2'-OMe) A, G, C, or U; Af, Gf, Cf, or Uf are 2'-fluoro dA, G, C, or U; dT is 2'-deoxythymidine; s is a phosphorothioate linkage. The double-stranded ribonucleic acid has a covalently bound triantennary GalNAc ligand.

[0135] In some embodiments, the RNAi agent is as shown in FIG.

[0136] RNAi agents can be in "naked" form or in the form of "free RNA." The embodiment refers to the absence of a pharmaceutical composition. For example, a naked RNAi agent can be, for example, Suitable glycerides may include citric acid, prolamin, carbonic acid, or phosphoric acid, or combinations thereof. In some embodiments, the buffer is phosphate buffered saline (PBS). The pH and osmolarity of the buffer containing the RNAi agent are adjusted so that it is suitable for administration to a subject. In certain embodiments, the RNAi agent may be in water for injection.

[0137] Alternatively, the RNAi agent may be administered as a pharmaceutical composition comprising the RNAi agent and one or more pharma- ceutically acceptable carriers. The pharmaceutical composition may be formulated based on the mode of delivery. The substance may be delivered parenterally, for example, by intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion. Alternatively, the compositions may be formulated for systemic administration by, for example, continuous pump infusion. The composition may be formulated for direct delivery into the brain parenchyma by intracerebral injection or Alternatively, the compositions can be administered orally; topically (e.g., by a transdermal patch); Pulmonary administration, for example, by inhalation or insufflation of powders or aerosols, including by nebulizer; The composition may be formulated for intranasal administration; intranasal administration; epidermal administration; or transdermal administration.

[0138] Compositions and formulations for parenteral, intraparenchymal (intracerebrovascular), intrathecal, intraventricular or intrahepatic administration may be buffered. agents, diluents, as well as penetration enhancers, carrier compounds and other pharma- ceutically acceptable carriers or excipients. The present invention may include, but is not limited to, a sterile solution which may also contain other suitable additives such as Compositions and formulations for oral administration may be in the form of powders or granules, microparticles, nanoparticles, Suspensions or solutions in aqueous or non-aqueous media, capsules, gel capsules, sachets, tablets or minitablets. Pharmaceutical compositions and formulations for topical administration may also be used. , transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays The pharmaceutical compositions of the present invention may include, but are not limited to, liquids and powders. These may also include, but are not limited to, solutions, emulsions, and liposome-containing formulations. The products are tablets, capsules, gel capsules, liquid syrups, soft gels, suppositories, and enemas. The formulation may be formulated in a variety of forms, including, but not limited to, enteral formulations.

[0139] Examples of pharma- ceutically acceptable carriers and specific compositions are described in PCT application Ser. No. 2016 / 013363, filed Aug. 25, 2016. No. PCT / US2016 / 048666 and U.S. Application No. 14 / 650,128, filed June 5, 2015. which are incorporated herein by reference.

[0140] In certain embodiments, the RNAi agent is administered at a dose of about 50 mg to about 800 mg, about 100 mg to about 800 mg. , about 150 mg to about 800 mg, about 200 mg to about 800 mg, 250 mg to about 800 mg, about 300 mg to about 800 mg, About 350 mg to about 800 mg, about 400 mg to about 800 mg, about 450 mg to about 800 mg, about 500 mg to about 800 mg, About 550 mg to about 800 mg, about 600 mg to about 800 mg, about 650 mg to about 800 mg, about 700 mg to about 800 mg, about 750 mg to about 800 mg, about 50 mg to about 750 mg, about 100 mg to about 750 mg, about 150 mg to about 750 mg, About 200 mg to about 750 mg, about 250 mg to about 750 mg, about 300 mg to about 750 mg, about 350 mg to about 750 mg, About 400 mg to about 750 mg, about 450 mg to about 750 mg, about 500 mg to about 750 mg, about 550 mg to about 750 mg, about 600 mg to about 750 mg, about 650 mg to about 750 mg, about 700 mg to about 750 mg, about 50 mg to about 700 mg, About 100 mg to about 700 mg, about 150 mg to about 700 mg, about 200 mg to about 700 mg, about 250 mg to about 700 mg, About 300 mg to about 700 mg, about 350 mg to about 700 mg, about 400 mg to about 700 mg, about 450 mg to about 700 mg, About 500 mg to about 700 mg, about 550 mg to about 700 mg, about 600 mg to about 700 mg, about 650 mg to about 700 mg, About 50 mg to about 650 mg, about 100 mg to about 650 mg, about 150 mg to about 650 mg, about 200 mg to about 650 mg, About 250 mg to about 650 mg, about 300 mg to about 650 mg, about 350 mg to about 650 mg, about 400 mg to about 650 mg, About 450 mg to about 650 mg, about 500 mg to about 650 mg, about 550 mg to about 650 mg, about 600 mg to about 650 mg, About 50 mg to about 600 mg, about 100 mg to about 600 mg, about 150 mg to about 600 mg, about 200 mg to about 600 mg, About 250 mg to about 600 mg, about 300 mg to about 600 mg, about 350 mg to about 600 mg, about 400 mg to about 600 mg, About 450 mg to about 600 mg, about 500 mg to about 600 mg, about 550 mg to about 600 mg, about 50 mg to about 550 mg, About 100 mg to about 550 mg, about 150 mg to about 550 mg, about 200 mg to about 550 mg, about 250 mg to about 550 mg, About 300 mg to about 550 mg, about 350 mg to about 550 mg, about 400 mg to about 550 mg, about 450 mg to about 550 mg, about 500 mg to about 550 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 150 mg to about 500 mg, About 200 mg to about 500 mg, about 250 mg to about 500 mg, about 300 mg to about 500 mg, about 350 mg to about 500 mg, about 400 mg to about 500 mg, about 450 mg to about 500 mg, about 50 mg to about 450 mg, about 100 mg to about 450 mg, About 150 mg to about 450 mg, about 200 mg to about 450 mg, about 250 mg to about 450 mg, about 300 mg to about 450 mg, about 350 mg to about 450 mg, about 400 mg to about 450 mg, about 50 mg to about 400 mg, about 100 mg to about 400 mg, About 150 mg to about 400 mg, about 200 mg to about 400 mg, about 250 mg to about 400 mg, about 300 mg to about 400 mg, about 350 mg to about 400 mg, about 50 mg to about 350 mg, about 100 mg to about 350 mg, about 150 mg to about 350 mg, about 200 mg to about 350 mg, about 250 mg to about 350 mg, about 300 mg to about 350 mg, about 50 mg to about 300 mg, About 100 mg to about 300 mg, about 150 mg to about 300 mg, about 200 mg to about 300 mg, or about 250 mg to about 300 mg. mg fixed dose, e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg g, about 225mg, about 250mg, about 275mg, 300mg, about 325mg, about 350mg, about 375mg, about 400mg, about 425mg , about 450mg, about 475mg, about 500mg, about 525mg, about 550mg, about 575mg, about 600mg, about 625mg, about 650mg g, about 675 mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg, or about 800 mg Values ​​and ranges intermediate to those listed above are also contemplated by the present invention. It is intended to be part of the light.

[0141] In some embodiments, the RNAi agent is about 1.0 ml, about 1.1 ml, about 1.2 ml, about 1.3 ml, about 1.4 ml , about 1.5 ml, about 1.6 ml, about 1.7 ml, about 1.8 ml, about 1.9 ml, or about 2.0 ml of pharmaceutical composition. The composition may be formulated at an appropriate concentration so that a sufficient volume of the composition is administered to the subject. In embodiments, the RNAi agent is administered to a subject in a volume of about 1.5 ml to provide a fixed dose of 300 mg of the agent. The compound is formulated in a suitable pharmaceutical formulation at about 200 mg / ml so as to

[0142] Unless otherwise defined, all technical and scientific terms used herein are understood to be within the meaning of the present invention. It has the same meaning as commonly understood by a person skilled in the art to which the invention pertains. All publications, patent applications, patents, and other references mentioned in the specification, as well as the Sequence Listing and Figures are incorporated by reference in their entirety. In case of conflict, the following claims, including definitions, are hereby incorporated by reference: In addition, the materials, methods, and examples are illustrative only and not limiting. It is not intended to do so. EXAMPLES

[0143] In patients with high cardiovascular risk and elevated LDL-C, a single or multiple subcutaneous injection A placebo-controlled, double-blind, randomized controlled trial to compare the effects of different doses of RNAi agents administered A randomized study was conducted.

[0144] method The RNAi agent has the nucleotide sequence 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO:3). The antisense strand of the sequence and the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO: 4) A double-stranded ribonucleic acid comprising a sense strand of the nucleotide sequence, wherein a, g, c and u are 2'-O-methyl ( 2'-OMe)A, G, C or U; Af, Gf, Cf or Uf is 2'-fluoro A, G, C or U; dT is 2'-deoxythymidine; s is a phosphorothioate linkage. , which had a covalently attached triantennary GalNAc ligand.

[0145] The primary objective was to evaluate the effect of RNAi agent treatment on LDL-C levels at day 180. Secondary objectives were to evaluate the effect of RNAi agents on: (i) 90-day (ii) LDL-C levels in the eye; (iii) PCSK9 levels over time; (iv) other lipids, lipoproteins, apolipoproteins; (v) pre-specified overall lipid guideline (vi) the proportion of patients achieving a baseline; (vi) individual responses to different doses; (vii) different (viii) the duration of lipid-lowering effect of the RNAi agent; and (viii) the safety and tolerability profile of the RNAi agent. The study also collected / assessed the effect of RNAi agents on: (a) cardiovascular death, non-fatal cardiac events; Cardiovascular events, including myocardial infarction, resuscitated cardiac arrest, and non-fatal stroke (ischemic and hemorrhagic) and (b) an anti-drug antibody against an RNAi agent.

[0146] This study included patients with ASCVD or conditions at risk for ASCVD (e.g., diabetes and familial hypercholesterolemia) The study included 501 patients with hypercalcaemia (hypercalcitriol) and elevated LDL-C. Subjects were included if they met all of the following inclusion criteria: (a) Male or female patients aged 18 years or older; (b) ASCVD or conditions at risk for ASCVD (symptomatic atherosclerosis, type 2 diabetes mellitus) , familial hypercholesterolemia, assessed by the Framingham risk score* or equivalent (including patients with a target LDL-C of <100 mg / dl) Medical history; (c) At screening, for ASCVD patients, serum LDL-C ≥ 1.8 mmol / l (≥ 70 mg / L) dl), or for patients at risk for ASCVD, serum LDL-C ≥ 2.6 mmol / l (≥ 100 mg / kg). / dl); (d) fasting triglycerides <4.52 mmol / l (<400 mg / dl) at screening; (e) Calculated by estimated glomerular filtration rate (eGFR) using standardized local clinical methodology glomerular filtration rate >30 ml / min; (f) Patients taking statins should be taking the maximally tolerated dose (clinical trials) investigator's judgment); (g) Patients receiving lipid-lowering therapy (e.g., statins and / or ezetimibe) should be screened. Patients should have been taking the medication steadily for at least 30 days prior to study initiation. There are no plans to change the (h) Willingness to give informed consent prior to the initiation of any study-related procedures. are able and willing to comply with any required testing procedures. *Framingham Risk Score >20%

[0147] Patients were excluded from the study if any of the following exclusion criteria applied immediately prior to randomization: Excluded from: (a) may interfere with participation in a clinical trial and / or if the patient has participated in a clinical trial , who may be exposed to significant risk (principal investigator [or delegate] in the judgment of the Department), uncontrolled or serious illness, or medical or surgical condition; (b) Any information that, in the opinion of the investigator (or sub-investigator), may interfere with the interpretation of the clinical trial results. known underlying illness or surgical, physical, or medical condition that may be causing the disease; (c) New York Heart Association (NYHA) class II, III, or IV heart failure or current known left ventricular Ejection fraction <30%; (d) cardiac arrhythmia within 3 months prior to randomization that was not controlled by medication or ablation; (e) history of any hemorrhagic stroke; (f) major adverse cardiac events within 6 months prior to randomization; (g) Uncontrolled severe hypertension: contraction before randomization despite antihypertensive therapy periodic blood pressure >180mmHg or diastolic blood pressure >110mmHg; (h) Inadequately controlled type 2 diabetes, i.e., glycated hemoglobin A1c ( HbA1c)>10.0%; (i) Defined as any known current infectious, neoplastic, or metabolic condition of the liver. Active liver disease confirmed by repeated measurements at least one week apart Alanine aminotransferase, aspartate aminotransferase, and ketamine aminotransferase, due to unknown causes, during cleaning Elevation of transferases >2x the ULN or total bilirubin >1.5x the ULN Rise; (j) severe comorbidities (e.g., acute systemic infection, or other serious illness); this includes treated Includes any cancer except basal cell carcinoma; (k) If pregnant, breastfeeding, or of possible pregnancy and using at least two methods of contraception (oral contraceptives) Contraceptives, barrier methods, approved contraceptive implants, long-term injectable contraceptives, intrauterine devices or Women who do not wish to use tubal ligation (tubal ligation)**; tubal ligation defined as more than 1 year since last menstrual period >2 years postmenopause and, if <55 years of age, a negative pregnancy test within 24 hours of randomization Certain women, or those who have been surgically sterilized, are exempt from this exclusion; (l) Use of an acceptable method of contraception (i.e., condoms with spermicide) throughout the study period Men who do not wish to use; (m) a known history of alcohol and / or drug abuse; (n) Exposure to another investigational drug or device within 30 days or 5 half-lives, whichever is longer. Treatment by; (o) Use of other investigational drugs or devices during the study; (p) Anything that would be of use to the investigator in conducting the study, including, but not limited to, Any conditions that may prevent this: (i) This trial, including patients who are unable to communicate or cooperate with the investigator; Unsuitable conditions for testing; (ii) Protocol requirements, instructions and limitations related to the study, the nature, scope and origin of the study; Inability to understand possible consequences (patients whose cooperation is questionable due to drug or alcohol abuse) (including); (iii) Are unlikely to comply with protocol requirements, instructions, and limitations associated with the study (e.g., Examples include uncooperative attitudes, failure to return for follow-up visits, and inability to complete the study. (unlikely condition); (iv) In the opinion of the investigator, participation in the study would increase the patient's risk Any medical or surgical condition that may be causing (v) someone directly involved in the conduct of the trial or a close relative is involved; (vi) any known cognitive impairment (e.g., Alzheimer's disease); and (q) Previous or current treatment with monoclonal antibodies against PCSK9 (9 of Screening) 0 days). **For the entire duration of the study

[0148] Patients were screened and randomized to one of six RNAi agent groups or a placebo group. Allocation was stratified by country and by current use of statins or other lipid-modifying therapies. Each patient received (i) one or two injections of an RNAi agent or placebo on day 1 only; or (ii) one injection on day 1 and two injections of an RNAi agent or placebo on day 90. I had an eye injection.

[0149] The study design is outlined in Figure 2. On day 1, all eligible patients were randomized The subjects received an initial subcutaneous dose of RNAi agent or placebo. The RNAi agent was administered as a single subcutaneous injection (dose: 200 mg, 3 The drug was administered either as a single injection (dose: 500 mg) or as two injections (dose: 500 mg). Patients randomized to receive a second injection of RNAi agent or placebo on day 90.

[0150] Placebo was administered as one or two subcutaneous injections of saline. The volume of placebo was Doses and volumes of RNAi agents were matched within injection regimens but not between injection regimens. For example, the 200 mg placebo group received 1.0 ml of placebo, whereas the 300 mg The placebo group received 1.5 ml of placebo.

[0151] The duration of a patient's involvement in a study may include screening, administration of the study drug, and single or multiple injections. The follow-up period, including the course of the injection and up to day 210, was approximately 224 days. The maximum duration of intervention, if required, was 374 days. End-of-study evaluations were performed on Day 210, except for patients who did not return within 24 hours of the end of study; these patients were assessed at 210 days and thereafter until 360 days or until LDL-C was within 20% of the starting level. Patients were followed up with clinic visits every 30 days until recovery (whichever occurred first).

[0152] In the single dose group (1 or 2 injections on day 1), patients followed the following schedule: (a) Screening: Day -14 to Day -1 (b) Randomization, Start of Study Drug: Day 1 (c) Treatment phase: Day 1 (d) Follow-up: (i) Follow-up: Days 2 to 210; EOS at Day 210 (ii) additional follow-up (patients whose LDL-C levels did not recover to >80% of baseline values) Patients will be re-evaluated monthly for follow-up until this level is reached or until day 360. Patients will visit the hospital on: days 240, 270, 300, 330, and 360.

[0153] In the two-dose group (one injection on days 1 and 90), patients were administered the following schedule: Went: (a) Screening: Day -14 to Day -1 (b) Randomization, Start of Study Drug: Day 1 (c) Treatment phase: Days 1 to 90 (d) Follow-up: (i) Follow-up: days 91 to 210; EOS at day 210 (ii) additional follow-up (patients whose LDL-C levels did not recover to >80% of baseline values) Patients will be re-evaluated monthly for follow-up until this level is reached or until day 360. Patients will visit the hospital on: days 240, 270, 300, 330, and 360.

[0154] The primary endpoint was the percentage change in LDL-C from baseline to day 180. It was evaluated by deciding

[0155] Secondary endpoints were (i) percentage change in LDL-C from baseline to day 90; (ii) From baseline to days 14, 30, 60, 120, 150, and 210 (iii) percentage change in LDL-C from baseline at days 180 and 210 (iv) the proportion of treated patients with an LDL-LDL concentration greater than 80% of baseline; -C or PCSK9 protein; (v) at days 90, 120, and 180, Treatment results in LDL-C levels of <25 mg / dl, <50 mg / dl, <70 mg / dl, and <100 mg / dl (vi) individual response, defined as the number of patients who achieved (vii) the proportion of patients in each group with a 50% or greater reduction in LDL-C from baseline; 14th, 30th, 60th, 90th, 104th, 120th, 150th, 180th day from Sline, and percentage change in PCSK9 levels through day 210; (viii) from each subsequent visit through day 210 Other lipids, lipoproteins, and apolipoproteins from baseline to (ix) achieving the overall lipid-modifying goal for the level of ASCVD risk The results were assessed by determining the proportion of patients in each group who

[0156] Efficacy was assessed using the following measures: total cholesterol, triglycerides, HDL-C, non-HDL-C, VLDL-C, and Apo- For LDL-C lipid and lipoprotein levels, including A1, Apo-B, Lp(a), CRP, and PCSK9 This included measuring the effect of RNAi agents on

[0157] Adverse events, serious adverse events, vital signs, clinical test values ​​(blood tests, coagulation tests, chemistry Clinical laboratory tests (including blood tests and urine tests) and electrocardiograms (ECGs) were collected at specific visits until the EOS visit (day 210). During additional monthly follow-up visits, adverse events, serious adverse events, and Laboratory tests continued to be evaluated (unless LDL-C levels had recovered to >80% of baseline). Cardiovascular events were cardiovascular death, non-fatal myocardial infarction, major coronary artery disease, and Events (death from CHD, resuscitated cardiac arrest, non-fatal myocardial infarction), ischemic stroke, and Summary of information on cardiovascular events, such as hemorrhagic stroke, reported as adverse events Ta.

[0158] In addition, anti-drug antibodies were assessed for RNAi agents. Anti-drug antibody formation was observed on day 1 (post-injection). before and 4 hours after), and on the 30th, 60th, 90th, 120th, 150th, and 180th days (150th and and 180 days only in patients who received a second dose of the RNAi agent), and at 210 days, The baseline LDL-C level was evaluated until the anti-drug antibody response became negative during the study period. For patients who have not yet reached above 80% of their normal LDL-C limit, Antidrug antibody formation was assessed at one follow-up visit at 1 year.

[0159] An independent Data Monitoring Committee (DMC) has confirmed that the first 40 patients will receive either the RNAi agent or placebo. Safety data were examined after the first injection and completion of the 14-day follow-up visit. The DMC then reviewed safety data every 2 months until the end of the study.

[0160] The primary endpoint was the least squares mean percentage from baseline to day 180 The changes were analyzed as changes in baseline and follow-up outcomes by study arm and scheduled visits. The randomized controlled trial was carried out using a repeated measures linear effects model, including the study arm and the interaction between study arm and scheduled visits. Analyses were performed with treatment at each visit as a fixed effect and patient as a random effect. Using the autoregressive variance structure embedded in the model, the PROC MIXED procedure was used in SAS software. Six RNAi agents were used for both the primary and secondary endpoints. Dunnett's test was used for comparisons between the treatment group and the placebo control group, and P values ​​were calculated for multiple comparisons. Separate doses were administered for each dosing strategy, i.e., single dose and two doses. The significance level for type I error in a two-sided test was 0.05.

[0161] All patients who received at least one dose of RNAi agent or placebo were included in the safety analysis. The pre-specified modified intention-to-treat population consisted of patients who had received at least one dose of the investigational drug were administered 10 mg / kg / day and both baseline and 180-day follow-up LDL-C level measurements were available. The intention-to-treat analysis was performed based on missing data. For patients with refractory disease, imputation was used.

[0162] Time course data are presented as means with 95% confidence intervals. Interpatient variability in response was The data are presented graphically using waterfall plots. The analysis was performed using SAS software. This was done using version 9.2 or higher (SAS Institute).

[0163] result Baseline demographics and medical history shown in Table 1 and Table 2 At the time of the study, 73% of patients were were receiving tinib therapy, and 31% were taking ezetimibe.

[0164] A single dose of RNAi agent on day 1 significantly increased IL-1 expression by 44.0% over baseline at day 30 across all doses. Resulted in a mean reduction in LDL-C ranging from 5% to 51.5% lower (see Figures 3 and 4), with the lowest reduction occurring at approximately day 60. As shown in FIG. 3, a single administration of a 200-mg dose of RNAi agent resulted in a significant increase in the number of tumors at 30 and 60 days. and reduced LDL-C by more than 40% from baseline at 90 days, and by 25% by 270 days. Single administration of 300-mg and 500-mg doses of RNAi agents was shown to be effective from day 15 to day 150. This treatment reduced LDL-C by more than 40% from baseline and by 30% by day 270 (Figure 3). For each RNAi agent dose intensity, LDL-C remained >20% below baseline through Day 360 The least squares mean reduction was greater for the RNAi agent than for the placebo (2.1% increase). The reduction in the number of single doses (27.9-41.9%) was significantly greater after the first dose (P<0.001 for all doses) (Ta Table 4. Mean LDL-C changes from baseline across the 200-mg, 300-mg, and 500-mg doses The average reduction was between 26% and 34.3% at 270 days and 30.2% to 32.2% at 360 days. In addition, the time-adjusted percentage changes in LDL-C between baseline and day 360 ranged between 0.01 and 0.18. Changes in LDL-C (i.e., changes in LDL-C at each time point measured between baseline and day 360) The mean percent change in DL-C was −31.6 for the 200-mg, 300-mg, and 500-mg doses, respectively. %, -38.1%, and -39.8% (see Figure 4). Interim-adjusted absolute change (i.e., LDL-C at each time point measured between baseline and day 360) The mean absolute change in LDL-C at the 200-mg, 300-mg, and 500-mg doses was -39. The responses of individual patients were: 1 mg / dl, -43.6 mg / dl, and -53.0 mg / dl (see Figure 5). The coefficient of variation for the change in LDL-C in individual patients receiving 100, 200, 300, and 500 mg was From baseline to day 270, the rates were 45.2%, 60.8%, and 50.7%, respectively. From the first day to the 360th day, the rates were 45.1%, 56.8%, and 50.2%, respectively (see Figures 6A to 6B).

[0165] Administration of a second dose of the RNAi agent on day 90 further reduced LDL-C compared to baseline. The mean reduction in LDL-C across all doses ranged from 34.2% to 44.1% at 90 days. The mean mean and mean mean ranged from 41.1% to 54.6% at day 1 and day 120 (see Figure 7). The difference between the patients who received two doses of the RNAi agent and placebo was significant: at day 180, The least squares mean reduction from baseline in LDL-C levels in subjects ranged from 35.5% to 52.6%. whereas the placebo group had an increase of 1.8% from baseline (total vs placebo P<0.001 for all comparisons (see Table 4). Treatment maintained LDL-C >30% and >40%, respectively, below baseline through day 210 (Figure 7 The second 300-mg dose was administered on days 120 through 210 to reduce LDL-C by >50% and baseline. Baseline across the 100-mg, 200-mg, and 300-mg doses The mean reduction in LDL-C from the study ranged from 25.6% to 43.4% at 270 days and 360% at 370 days. The mortality rates ranged from 13.3% (100-mg dose) to 33.3% (300-mg dose). The time-adjusted percent change in LDL-C between day 1 and day 360 was 0.01–0.25 at 200-mg, 300-mg, and 50-mg doses. The changes in the 0-mg dose were -31.0%, -41.2%, and -46.8%, respectively (see Figure 4). Time-adjusted absolute change in LDL-C between 0 and 60 days was 0.01 for the 200-mg, 300-mg, and 500-mg doses. The mean blood glucose levels in each patient were -39.6 mg / dl, -39.1 mg / dl, and -57.7 mg / dl, respectively (see Figure 5). Responses are shown in Figures 8 and 9. At Day 180, mean LDL-C from baseline The reduction was 52.6%, with a maximum of 81% (see Figure 8). 100 mg, 200 mg, and 300 mg were administered. The coefficient of variation for the change in LDL-C in patients from baseline to day 270 was 45.7 and 46.7, respectively. %, 82.3%, and 73.7%, and from baseline to day 360, the rates were 46.0%, 77.5%, and 78.2%, respectively. , and 74.2% (see FIGS. 9A to 9B).

[0166] In patients receiving placebo against a background of the maximum available dose of statin, There was considerable variation in the change in LDL cholesterol levels from baseline at 180 days. (mean [±SD] absolute difference, −0.7 ± 25.6 mg per deciliter [−0.02 ± 0.66 mg per liter] In contrast, all patients who received two 300-mg doses of RNAi agent showed The mean absolute change in LDL cholesterol levels per deciliter was 1.04 mg / dL. The mean mean blood glucose concentration was −64.2 ± 20.7 mg [−1.66 ± 0.54 mmol per liter] (see Figure 10B), and 54% of patients had a mean blood glucose concentration of 50% or more. In this RNAi agent dose group, 5%, 48%, and 66% of patients had a 1-dose reduction at day 180. Less than 25 mg per liter (0.65 mmol per liter), 50 mg per deciliter ( Less than 1.3 mmol per torr (1.3 mmol per liter) and less than 70 mg per deciliter (1.8 mmol per liter) At day 240, the individual cholesterol levels in the same patient groups were Terrol levels remained lower than at baseline (see FIG. 10C).

[0167] Comparison of individual patient data between administration of a single dose of 300 mg and administration of two doses of 300 mg showed Administration of 300 mg of RNAi agent on days 1, 90, 270, and every 6 months thereafter was The authors suggest that this would reduce variability and result in a sustained reduction in LDL-C by approximately 50%. (See FIGS. 11A and 11B).

[0168] There was also a significant reduction in the levels of non-HDL-C and Apo-B, but not high-sensitivity C-reactive protein (HSC). The levels of The percentage changes from baseline for additional lipid measurements are shown in Table 3(Table 3) to Table 5(Table 5).

[0169] As shown in Figures 12 and 13, after the RNAi agent was administered on day 1, the Across the range of RNAi agent doses, PCSK9 levels decreased by an average of 59.6% and 68% from baseline levels. At day 30, PCSK9 levels were reduced by 66.2% below baseline levels. The RNAi agent further reduced the expression of β-glucose (β-glucose) by between 10 and 74.0%, with similar reductions occurring at 60 and 90 days. In patients who received a single dose, the mean reduction in PCSK9 levels at 180 days ranged from 47.9% to 59. The mean mean mean scores ranged between 0.3% and 0.3% (P<0.001 for each dose vs. placebo) (see Table 4). In patients who received a single 500-mg dose, PCSK9 levels remained 60% above baseline by day 150. In contrast, in patients who received two doses of the RNAi agent, the second Further reductions in PCSK9 levels were observed after 40 days of treatment. At day 180, there was a 7.0% to 62.8% reduction, and at day 120 there was a 60.4% to 74.5% reduction. Mean increase from baseline in PCSK9 levels in patients receiving two doses of an RNAi agent in The reductions ranged between 53.2% and 69.1% (P<0.001 for each dose vs. placebo) (Table 4 (see Table 4). In patients who received two doses of 200 mg or 300 mg, PCSK9 levels remained stable through 240 days. Remained greater than 60% below baseline. In relation to the regimen, the reduction in PCSK9 levels at day 270 was greater than about 40% (Figure 12 and (See Figure 13.)

[0170] To predict LDL-C levels over time when an RNAi agent is administered at 300 mg twice or three times a year A model was developed and verified. The model was based on observed and predicted data up to 210 days. The results at 270 days were consistent with those at 210 days of follow-up. The pharmacodynamics model using data from patients who underwent Figure 1 shows modeled LDL-C levels over the course of months.

[0171] At 210 days, 76% of patients receiving the RNAi agent and 76% of patients receiving the placebo Adverse events were reported (Table 7; see also Table 6). Most of these events (95%) The incidence of serious adverse events was 1.1% in the RN group and 1.2% in the RN group. The incidence rates were 11% in patients receiving Ai and 8% in patients receiving placebo. Two patients died due to adverse events. One patient discontinued the study due to a herpes zoster infection (placebo group) and the other due to influenza. The most common adverse events were influenza or nasopharyngitis (two doses of 100 mg RNAi agent group). (occurring in >2% of patients) are myalgia, headache, fatigue, nasopharyngitis, back pain, high blood pressure, diarrhea, and dizziness. The incidence of these events was significantly higher between the groups receiving the RNAi agent and the group receiving the placebo. There was no doubt about it.

[0172] Injection site reactions occurred in 4% of patients who received a single dose of the RNAi agent and in 7% of patients who received two doses ( after one or both doses (pooled rate 5%); injection site reactions occurred more frequently than placebo None occurred in assigned patients (Table 7; Table 8 and Table 9). reference).

[0173] Two patients had elevated levels of hepatic aspartate aminotransferase ( >3x the upper limit of normal), 1 in the single-dose placebo group, and 1 in the single-dose 300-mg RNAi Patients in the 300-mg RNAi group had elevated hepatic alanine aminotransferase levels. Two additional patients (one in the two-dose 100 mg RNAi agent group and one in the two-dose 3 One patient in the 0.00 mg RNAi group also had elevated alanine aminotransferase levels. All aminotransferase elevations were transient. In patients with bilirubin levels above 100 mg / kg, there were no significant differences in bilirubin levels associated with the RNAi agent or placebo. There was no increase in lirubin levels, and the liver cell damage was accompanied by jaundice (not a cholestatic disorder). Patients are at high risk for fatal drug-induced liver injury if given a drug that causes None of the cases met the definition of Hy's law. One patient had received antimicrobial therapy before the first injection. All patients were positive for anti-drug antibodies; no other cases of anti-drug antibodies have been reported (Table 10). (See Table 10).

[0174] There were no additional safety findings at follow-up from days 210 to 360 (T (See Table 11).

[0175] Two deaths occurred late in the trial. The first was in a patient with long-term vascular disease and frequent angina. The study occurred in patients randomly assigned to the 500-mg single-dose RNAi agent group, who had previously received the drug. The patient presented with cardiac arrest and died on day 104. The second death occurred after a percutaneous repair following study enrollment. A man in the two-dose 200-mg RNAi group who had a thoracic aortic aneurysm and subsequently developed a fistula and sepsis he died on day 198 of the trial.

[0176] conclusion These results demonstrate that RNAi agents significantly reduce LDL-C in patients over a period of several months. Treatment with RNAi agents significantly increased Lp(a) and total cholesterol, among other lipid parameters. The RNAi agents acted against PCSK9, LDL-C, non-HDL-C, and Apo-B. A dose-response effect was observed. The variability of LDL-C in individual patients was virtually eliminated, and The effect was sustained between doses.

[0177] In addition, administration of RNAi agents is well tolerated by patients and is not associated with significant safety issues. TEAEs were rare and mild to moderate, with minimal injection burden. Ta.

[0178] Overall, RNAi agents offer the opportunity to improve patient adherence.

[0179] [Table 1A]

[0180] [Table 1B]

[0181] [Table 1C]

[0182] [Table 2]

[0183] [Table 3]

[0184] [Table 4A]

[0185] [Table 4B]

[0186] [Table 5]

[0187] [Table 6A]

[0188] [Table 6B]

[0189] [Table 7A]

[0190] [Table 7B]

[0191] [Table 8]

[0192] [Table 9]

[0193] [Table 10]

[0194] [Table 11]

[0195] Thus, the embodiments of the present invention have been described in detail. The invention is susceptible to many obvious variations thereof without departing from the spirit or scope of the invention. It will be understood that the present invention should not be limited to the specific details set forth in the above description, as various modifications and variations are possible within the spirit and scope of the present invention. This is what should be done. [Sequence List Free Text]

[0196] SEQ ID NO:1 5'-ACAAAAGCAAAACAGGUCUAGAA-3' SEQ ID NO:2 5'-CUAGACCUGUTUUGCUUUUGU-3' SEQ ID NO:3 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3', where a, g, c and u are 2'-O-methyl Af, Gf, Cf or Uf is 2'-fluoro A, G, C or U; dT is 2'-deoxythymidine; s is a phosphorothioate linkage SEQ ID NO:4 5'-csusagacCfuGfudTuugcuuuugu-3', where a, g, c and u are 2'-O-methyl (2'-OMe)A Af, Gf, Cf or Uf is 2'-fluoro A, G, C or U; dT is 2'-de oxythymidine; s is a phosphorothioate linkage

Claims

1. A method for lowering low density lipoprotein cholesterol in a subject, comprising the steps of: administering to a subject an effective amount of an interfering ribonucleic acid (RNAi) agent, RNAi is a double-stranded ribonucleic acid that contains a sense strand and an antisense strand that form a double-stranded region. The antisense strand comprises the nucleotide sequence of SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:

4. The method comprises the step of:

2. A method for preventing a cardiac event in a subject, comprising administering to a subject a therapeutically effective amount of interfering ribonucleic acid (RNA i) administering the agent to a subject; RNAi is a double-stranded ribonucleic acid that contains a sense strand and an antisense strand that form a double-stranded region. The antisense strand comprises the nucleotide sequence of SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:

4. The method comprises the step of:

3. A method for reducing cardiovascular mortality and morbidity in a subject, comprising administering to a subject a therapeutically effective amount of an interfering agent. administering a ribonucleic acid (RNAi) agent to a subject; RNAi is a double-stranded ribonucleic acid that contains a sense strand and an antisense strand that form a double-stranded region. The antisense strand comprises the nucleotide sequence of SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:

4. The method comprises the step of:

4. The subjects were those with atherosclerotic cardiovascular disease (ASCVD), conditions at risk for ASCVD, and those with heart disease. Increased risk of vascular disease, heterozygous familial hypercholesterolemia or homozygous have familial hypercholesterolemia or need to lower LDL-C, or The method according to any one of claims 1 to 3, comprising a combination thereof.

5. The subject has a baseline low-density lipoprotein (LDL-C) level of about 70 mg / dl or greater.

5. The method according to any one of claims 1 to 4.

6. 6. The method of any one of claims 1 to 5, wherein the subject does not have active liver disease.

7. Active liver disease is characterized by: alanine aminotransferase (ALT) levels greater than 2x the upper limit of normal (ULN); aspartate aminotransferase (ALT); aspartate aminotransferase (AST) greater than twice the ULN; and UL The method according to claims 1 to 6, characterized by one or more of the following: total bilirubin greater than 1.5 times N 2. The method according to any one of claims 1 to 11.

8. 8. The method of any one of claims 1 to 7, wherein the subject has active liver disease.

9. Active liver disease is characterized by: alanine aminotransferase (ALT) levels greater than 2x the upper limit of normal (ULN); aspartate aminotransferase (ALT); aspartate aminotransferase (AST) greater than twice the ULN; and UL The method of claim 8, characterized by one or more of the following: total bilirubin greater than 1.5 times N How to.

10. 10. The method of claim 1, wherein the subject is being treated with background lipid-lowering therapy. The method according to claim 5.

11. Background lipid-lowering therapy consisted of statins, ezetimibe, and LDL apheresis. The method of claim 10, wherein the .alpha.-amino acid is selected from the group consisting of .alpha.-amino acid and .beta.-amino acid.

12. 12. The method of claim 10 or 11, wherein the background lipid-lowering therapy is a statin.

13. 13. The method according to claim 10, wherein the subject is on maximally tolerated statin therapy. The method according to claim 5.

14. Background lipid-lowering therapy is maintained while the subject is administered the RNAi agent. The method according to any one of claims 10 to 13.

15. 10. The method of claim 1, wherein the subject is not receiving background lipid-lowering therapy. The method according to

16. 1. The method of claim 1, wherein the subject has a baseline triglyceride level of about 400 mg / dl or less.

6. The method of any one of claims 5 to 5.

17. The subject has a baseline estimated glomerular filtration rate (eGFR) of at least about 30 ml / min.

17. The method according to any one of items 1 to 16.

18. 18. The method of claim 1, wherein the subject does not have poorly controlled type 2 diabetes. The method described.

19. Inadequately controlled type 2 diabetes with baseline glycated hemoglobin A1c of at least approximately 10% The method of claim 18, wherein the level is determined by

20. Subjects do not have New York Heart Association (NYHA) class II, III, or IV heart failure.

20. The method according to any one of claims 1 to 19.

21. 21. The method according to any one of claims 1 to 20, wherein the subject's last known ventricular ejection fraction is 30% or greater. Method of posting.

22. 1. The method of claim 1, wherein the subject has not experienced a major adverse cardiac event within 6 months of administration of the RNAi agent.

22. The method according to any one of claims 1 to 21.

23. 23. The method of any one of claims 1 to 22, wherein the subject has not experienced a hemorrhagic stroke. 。

24. 24. The method of claim 1, wherein the subject does not have cardiac arrhythmia within 3 months of administration of the RNAi agent.

3. The method according to claim 1 .

25. Subjects were not controlled by medication or resection within 3 months of administration of the RNAi agent.

25. The method of any one of claims 1 to 24, wherein the patient is free of cardiac arrhythmia.

26. 26. The method of any one of claims 1 to 25, wherein the subject does not have cardiac arrhythmia.

27. 27. The method of any one of claims 1 to 26, wherein the subject is an adult.

28. The subject is a patient with heterozygous familial hypercholesterolemia or homozygous familial hypercholesterolemia. The method of any one of claims 1 to 27, wherein the patient has choleremia.

29. 29. The method of any one of claims 1 to 28, wherein the subject is in need of lowering LDL-C.

30. Administration of an RNAi agent increases levels of LDL-C by approximately 20% or more compared to baseline LDL-C levels.

30. The method of claim 1, wherein the method significantly reduces

31. A reduction in the level of LDL-C of greater than about 20% compared to baseline levels is observed when the RNAi agent is administered.

31. The method of claim 1, wherein the method is maintained for 15 days or more after administration.

32. Administration of an RNAi agent increased PCSK9 levels by approximately 25% compared to baseline levels of PCSK9. and the baseline level of PCSK9 is measured prior to administration of the RNAi agent.

31. A method according to any one of claims 31 to 31.

33. A reduction in the level of PCSK9 of greater than about 25% compared to the baseline level is observed after administration of the RNAi agent. The method of any one of claims 1 to 32, wherein the method is maintained for 30 days or more after administration.

34. Cardiac events were death, non-fatal myocardial infarction, severe recurrent ischemia, stroke, symptomatic pulmonary 34. The method according to claim 2, wherein the vascular endothelial cell is selected from the group consisting of: embolism, and hemorrhage. How to.

35. 35. The method of claim 1, wherein the administration of the RNAi agent comprises a loading phase and a maintenance phase. The method according to claim 5.

36. 35. The method of claim 34, wherein the loading phase comprises administering the RNAi agent as one or more doses. The method according to

37. 37. The method of claim 36, wherein the loading phase comprises administering the RNAi agent as at least two doses. The method described.

38. 38. The method of claim 37, wherein at least two of the doses are separated by a time interval.

39. 39. The method of claim 38, wherein the time interval is from about 1 day to about 180 days.

40. 40. The method of claim 38 or 39, wherein the time interval is from about 60 days to about 120 days.

41. 41. The method of claim 38, wherein at least two doses are separated by a time interval of about 90 days. The method according to any one of claims 1 to 5.

42. 42. The method of any one of Claims 36 to 41, wherein the dose comprises about 50 to about 800 mg of the RNAi agent. 。

43. 43. The method of claim 42, wherein the dose comprises about 100 to about 500 mg of the RNAi agent.

44. 44. The method of claim 42 or 43, wherein the dose comprises about 300 mg of the RNAi agent.

45. 35. The method of claim 34, wherein the maintenance phase comprises administering the RNAi agent as one or more doses.

45. The method of claim 44,

46. 45. The method of claim 45, wherein the loading phase comprises administering the RNAi agent as at least two doses. The method according to

47. 47. The method of claim 46, wherein at least two of the doses are separated by a time interval.

48. 48. The method of claim 47, wherein at least two of the doses are separated by a regular time interval.

49. 49. The method of claim 48, wherein the regular time interval is between about 1 month and about 12 months.

50. 50. The method of claim 48 or 49, wherein the regular time interval is between about 3 months and about 9 months.

51. 51. The method of any one of claims 48 to 50, wherein the regular time interval is about 6 months.

52. 52. The method of any one of Claims 45 to 51, wherein the dose comprises about 50 to about 800 mg of the RNAi agent. 。

53. 53. The method of claim 52, wherein the dose comprises about 100 to about 500 mg of the RNAi agent.

54. 54. The method of claim 52 or 53, wherein the dose comprises about 300 mg of the RNAi agent.

55. A method for preventing the development of atherosclerotic cardiovascular disease in a subject, comprising: administering to a subject a therapeutically effective amount of an interfering ribonucleic acid (RNAi) agent, RNAi is a double-stranded ribonucleic acid that contains a sense strand and an antisense strand that form a double-stranded region. The antisense strand comprises the nucleotide sequence of SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:

4. The method comprises the step of:

56. ASCVD, ASCVD and risk-equivalent conditions, heterozygous familial hypercholesterolemia, homozygous People with conjugated familial hypercholesterolemia, who need to lower LDL-C, or A method of treating a subject having these combinations comprising administering a therapeutically effective amount of interfering ribonucleic acid (RIB) to a subject having the above-mentioned combination. Administering a NAi) agent to a subject, RNAi is a double-stranded ribonucleic acid that contains a sense strand and an antisense strand that form a double-stranded region. The antisense strand comprises the nucleotide sequence of SEQ ID NO:3, and the sense strand comprises the nucleotide sequence of SEQ ID NO:

4. The method comprises the step of:

57. 57. The method of claim 55 or 56, wherein the subject has a baseline LDL-C level of about 70 mg / dl or greater. method.

58. 58. The method of any one of claims 55 to 57, wherein the subject has active liver disease.

59. Active liver disease is characterized by: alanine aminotransferase (ALT) levels greater than 2x the upper limit of normal (ULN); aspartate aminotransferase (ALT); aspartate aminotransferase (AST) greater than twice the ULN; and UL The method according to claim 58, characterized by one or more of the following: total bilirubin greater than 1.5 times N Method of posting.

60. 58. The method of any one of claims 55 to 57, wherein the subject does not have active liver disease.

61. Active liver disease is characterized by: alanine aminotransferase (ALT) levels greater than 2x the upper limit of normal (ULN); aspartate aminotransferase (ALT); aspartate aminotransferase (AST) greater than twice the ULN; and UL The method according to claim 60, characterized by one or more of the following: total bilirubin greater than 1.5 times N Method of posting.

62. 62. The method of claim 55, wherein the subject is being treated with background lipid-lowering therapy. The method according to any one of claims 1 to 5.

63. Background lipid-lowering therapy consisted of statins, ezetimibe, and LDL apheresis.

63. The method of claim 62, wherein the

64. Background lipid-lowering therapy is maintained while the subject is administered the RNAi agent. The method according to claim 62 or 63.

65. 62. Any one of claims 55 to 61, wherein the subject is not receiving background lipid-lowering therapy. The method according to claim 5.

66. 55 to 59. The method of claim 55, wherein the subject has a baseline triglyceride level of greater than about 400 mg / dl.

65. The method of claim 65.

67. 55 to 59. The method of claim 54, wherein the subject has a baseline estimated glomerular filtration rate (eGFR) of less than about 30 ml / min.

67. The method of any one of claims 66 to 66.

68. 68. The method of any one of claims 55 to 67, wherein the subject has poorly controlled type 2 diabetes. How to.

69. Inadequately controlled type 2 diabetes with baseline glycated hemoglobin A1c of at least approximately 10% 69. The method of claim 68, wherein the level is identified.

70. The subject has New York Heart Association (NYHA) class II, III, or IV heart failure.

70. The method according to any one of paragraphs 55 to 69.

71. 71. The method of any one of claims 55 to 70, wherein the subject has a ventricular ejection fraction of less than 30%.

72. 55. The subject experiences a major adverse cardiac event within 6 months of administration of the RNAi agent.

72. A method according to any one of claims 1 to 71.

73. 73. The method of claim 55, wherein the subject has experienced at least one hemorrhagic stroke. The method described in paragraph .

74. 74. The method of claim 55, wherein the subject has a cardiac arrhythmia within 3 months of administration of the RNAi agent. The method according to claim 5.

75. Subjects were not controlled by medication or resection within 3 months of administration of the RNAi agent.

75. The method of any one of claims 55 to 74, wherein the patient is free of cardiac arrhythmia.

76. 76. The method of any one of claims 55 to 75, wherein the subject does not have cardiac arrhythmia.

77. 77. The method of any one of claims 55 to 76, wherein the subject is an adult.

78. The subject is a patient with heterozygous familial hypercholesterolemia or homozygous familial hypercholesterolemia. The method of any one of claims 55 to 77, wherein the patient has choleremia.

79. 79. The method of any one of claims 55 to 78, wherein the subject is in need of lowering LDL-C.

80. Low density lipoprotein was reduced by more than about 20% compared to baseline levels, and RNA 80. The method of any one of claims 55 to 79, wherein the i-agent is administered for 15 days or more. method.

81. Administration of an RNAi agent increased PCSK9 levels by approximately 25% compared to baseline levels of PCSK9. and wherein the baseline level of PCSK9 is measured prior to administration of the RNAi agent.

80. A method according to any one of claims 7 to 80.

82. A reduction in the level of PCSK9 of greater than about 25% compared to the baseline level is observed after administration of the RNAi agent.

82. The method of claim 81, wherein the method is maintained for 30 days or more after administration.

83. 83. The method of claim 55, wherein the administration of the RNAi agent comprises a loading phase and a maintenance phase. The method according to claim 5.

84. 83. The method of claim 82, wherein the loading phase comprises administering the RNAi agent as one or more doses. The method described above.

85. 84. The method of claim 83, wherein the loading phase comprises administering the RNAi agent as at least two doses. The method described.

86. 86. The method of claim 85, wherein the at least two doses are separated by a time interval.

87. 87. The method of claim 86, wherein the time interval is from about 1 day to about 180 days.

88. 88. The method of claim 86 or 87, wherein the time interval is from about 60 days to about 120 days.

89. 89. The method of any one of claims 86 to 88, wherein the time interval is about 90 days.

90. 90. The method of any one of Claims 83-89, wherein the dose comprises from about 50 to about 800 mg of the RNAi agent. 。

91. 91. The method of claim 90, wherein the dose comprises about 100 to about 500 mg of the RNAi agent.

92. 92. The method of claim 90 or 91, wherein the dose comprises about 300 mg of the RNAi agent.

93. 83. The method of claim 82, wherein the maintenance phase comprises administering the RNAi agent as one or more doses.

92. A method according to any one of claims 1 to 92.

94. 93. The method of claim 93, wherein the loading phase comprises administering the RNAi agent as at least two doses. The method described.

95. 95. The method of claim 94, wherein at least two of the doses are separated by a time interval.

96. 96. The method of claim 95, wherein the at least two doses are separated by a regular time interval.

97. 97. The method of claim 96, wherein the regular time interval is between about 1 month and about 2 months.

98. 98. The method of claim 96 or 97, wherein the regular time interval is between about 3 months and about 9 months.

99. 99. The method of any one of claims 96 to 98, wherein the regular time interval is about 6 months.

100. 100. The method of any one of Claims 93-99, wherein the dose comprises from about 50 to about 800 mg of the RNAi agent. 。

101. 101. The method of claim 100, wherein the dose comprises about 100 to about 500 mg of the RNAi agent.

102. 102. The method of claim 100 or 101, wherein the dose comprises about 300 mg of the RNAi agent.

103. Dietary therapy designed to improve lipid levels and maximally tolerated statin therapy A method for preventing a cardiac event in a subject receiving Patients with familial hypercholesterolemia who require further reduction of low-density lipoprotein cholesterol We need the following: The method includes administering a prophylactically effective amount of an RNAi agent, RNAi agents are double-stranded ribonucleic acids that contain a sense strand and an antisense strand that form a double-stranded region. the antisense strand comprises the nucleotide sequence of SEQ ID NO:3 and the sense strand comprises the nucleotide sequence of SEQ ID NO:4 The method comprises the steps of:

104. Dietary therapy designed to improve lipid levels and maximally tolerated statin therapy A method for preventing a cardiac event in a subject receiving Patients with atherosclerotic cardiovascular disease who require further reduction in low-density lipoprotein cholesterol It is essential to The method includes administering a prophylactically effective amount of an RNAi agent, RNAi agents are double-stranded ribonucleic acids that contain a sense strand and an antisense strand that form a double-stranded region. the antisense strand comprises the nucleotide sequence of SEQ ID NO:3 and the sense strand comprises the nucleotide sequence of SEQ ID NO:4 The method comprises the steps of:

105. Dietary and low-density lipoprotein lowering therapy designed to improve lipid levels A method for preventing a cardiac event in a subject receiving Patients with chronic hypercholesterolemia who require further reduction of low-density lipoprotein cholesterol We need it. The method includes administering a prophylactically effective amount of an RNAi agent, RNAi agents are double-stranded ribonucleic acids that contain a sense strand and an antisense strand that form a double-stranded region. the antisense strand comprises the nucleotide sequence of SEQ ID NO:3 and the sense strand comprises the nucleotide sequence of SEQ ID NO:4 The method comprises the steps of:

106. Dietary therapy designed to improve lipid levels and maximally tolerated statin therapy A method of treating a subject suffering from heterozygous familial hypercholesterolemia, have hypercalcemia and require further reduction in low-density lipoprotein cholesterol, The method includes administering a therapeutically effective amount of an RNAi agent, RNAi agents are double-stranded ribonucleic acids that contain a sense strand and an antisense strand that form a double-stranded region. the antisense strand comprises the nucleotide sequence of SEQ ID NO:3 and the sense strand comprises the nucleotide sequence of SEQ ID NO:4 The method comprises the steps of:

107. Dietary therapy designed to improve lipid levels and maximally tolerated statin therapy The present invention relates to a method for treating a subject suffering from atherosclerotic cardiovascular disease, the subject being and in need of further reduction in low density lipoprotein cholesterol, The method includes administering a therapeutically effective amount of an RNAi agent, RNAi agents are double-stranded ribonucleic acids that contain a sense strand and an antisense strand that form a double-stranded region. the antisense strand comprises the nucleotide sequence of SEQ ID NO:3 and the sense strand comprises the nucleotide sequence of SEQ ID NO:4 The method comprises the steps of:

108. Dietary and low-density lipoprotein lowering therapy designed to improve lipid levels A method of treating a subject having homozygous familial hypercholesterolemia, have a condition that requires further reduction in low-density lipoprotein cholesterol, The method includes administering a therapeutically effective amount of an RNAi agent, RNAi agents are double-stranded ribonucleic acids that contain a sense strand and an antisense strand that form a double-stranded region. the antisense strand comprises the nucleotide sequence of SEQ ID NO:3 and the sense strand comprises the nucleotide sequence of SEQ ID NO:4 The method comprises the steps of:

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