A method for preventing cardiovascular events by reducing the proprotein convertase subtilisin kexin 9 (PCSK9) protein.

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 claims the benefit of U.S. Provisional Patent Application No. 62 / 409,816, filed October 18, 2016; U.S. Provisional Patent Application No. 62 / 422,028, filed November 14, 2016; U.S. Provisional Patent Application No. 62 / 472,525, filed March 16, 2017; and U.S. Provisional Patent Application No. 62 / 550,426, filed August 25, 2017, the entire contents of each of which are incorporated herein by reference. This application also relates to U.S. Provisional Patent Application No. 62 / 209,526, filed August 25, 2015; PCT Application No. PCT / US2016 / 048666, filed August 2 5, 2016; U.S. Provisional Application No. 61 / 733,518, filed December 5, 2012; U.S. Provisional Application No. 61 / 793,530, filed March 15, 2013; U.S. Provisional Application No. 61 / 886,916, filed October 4, 2013; U.S. Provisional Application No. 61 / 892,

[0002] 188, filed October 17, 2013; PCT Application No. PCT / US2013 / 073349, filed December 5, 2013; and U.S. Patent Application No. 14 / 650,128, filed June 5, 2015, the entire contents of each of these provisional patent applications and PCT applications are incorporated herein by reference. 5, 2016; U.S. Provisional Application No. 61 / 733,518, filed December 5, 2012; U.S. Provisional Application No. 61 / 793,530, filed March 15, 2013; U.S. Provisional Application No. 61 / 886,916, filed October 4, 2013; U.S. Provisional Application No. 61 / 892, 188, filed October 17, 2013; PCT Application No. PCT / US2013 / 073349, filed December 5, 2013; and U.S. Patent Application No. 14 / 650,128, filed June 5, 2015, the entire contents of each of these provisional patent applications and PCT applications are incorporated herein by reference. 188, filed October 17, 2013; PCT Application No. PCT / US2013 / 073349, filed December 5, 2013; and U.S. Patent Application No. 14 / 650,128, filed June 5, 2015, the entire contents of each of these provisional patent applications and PCT applications are incorporated herein by reference. 188, filed October 17, 2013; PCT Application No. PCT / US2013 / 073349, filed December 5, 2013; and U.S. Patent Application No. 14 / 650,128, filed June 5, 2015, the entire contents of each of these provisional patent applications and PCT applications are incorporated herein by reference. This application is also related to U.S. Provisional Patent Application No. 62 / 209,526, filed August 25, 2015; PCT Application No. PCT / US2016 / 048666, filed August 2 5, 2016; U.S. Provisional Application No. 61 / 733,518, filed December 5, 2012; U.S. Provisional Application No. 61 / 793,530, filed March 15, 2013; U.S. Provisional Application No. 61 / 886,916, filed October 4, 2013; U.S. Provisional Application No. 61 / 892,

[0003] Sequence Listing This application contains a sequence listing submitted electronically in ASCII format, the entire contents of which are incorporated herein by reference. The ASCII copy created on October 18, 2017 is named A115- 2_3_SL.txt and is 1,799 bytes in size. 2_3_SL.txt and is 1,799 bytes in size.

Background Art

[0004] Atherosclerotic cardiovascular disease (ASCVD) remains a global health challenge. It is characterized by the accumulation of fatty deposits, inflammatory cells, and scar tissue within the arterial wall. Atherosclerosis, a disease process, is the underlying cause of the majority of clinical cardiovascular events. It is the underlying cause. Various types of treatments and preventive measures have been developed, but low-density lipoprotein Reducing protein cholesterol (LDL-C) remains a well-established strategy for combating ASCVD. This is a strategy. For this purpose, statins lower lipid levels. Although it has become a widely used treatment, many subjects are at risk, and the risk is rising. It continues to have a high level of LDL-C.

[0005] Proprotein convertase subtilisin kexin 9 PCSK9 (n 9) was recently identified as a target for reducing LDL-C levels. PCSK9 is It is a member of the butyl sine serine protease family and is involved in cholesterol metabolism. It has been shown that it will fulfill its role.

[0006] Monoclonal antibodies that block PCSK9 have been developed, reducing circulating PCSK9 levels. It was demonstrated that it lowers LDL-C levels. However, the monoclonal antibody of PCSK9 is ineffective. This is a short-term treatment, which can lead to significant administration and financial burdens.

[0007] Therefore, the goal is to reduce LDL-C levels and ultimately treat or prevent ASCVD. Effective therapeutic or preventive measures are needed in this field for the purpose of [this]. [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 No. 14 / 650,128 [Overview of the Initiative] [Problems that the invention aims to solve]

[0009] The present invention relates to a combination of PCSK9 in a preventive or therapeutic method for subjects requiring it. Regarding the use of RNA interference (RNAi) agents that inhibit growth. [Means for solving the problem]

[0010] In one embodiment, the present invention relates to a method for reducing LDL-C in a subject. This includes a step of administering an effective amount of RNAi agent to a target, where the RNAi agent forms a double-stranded region. It is a double-stranded ribonucleic acid containing a sense strand and an antisense strand. The antisense strand is sequence number It contains the nucleotide sequence of No. 3, and the sense strand contains the nucleotide sequence of SEQ ID NO: 4.

[0011] In another aspect, the present invention relates to a method for preventing cardiovascular events in a subject. This method includes the step of administering to the subject an effective amount of an RNAi agent, wherein the RNAi agent is a double-stranded ribonucleic acid containing 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 .

[0012] In yet another aspect, the present invention relates to a method for reducing the cardiovascular mortality and / or morbidity in a subject. This method includes the step of administering to the subject an effective amount of an RNAi agent, wherein the RNAi agent is a double-stranded ribonucleic acid containing 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 of. otide sequence.

[0013] In certain embodiments, the subject may have ASCVD, a condition with equivalent risk to ASCVD (ASCVD risk equivalent), an increased risk of cardiovascular disease (CVD), heterozygous familial hypercholesterolemia , or homozygous familial hypercholesterolemia, and there is a need to lower low-density lipoprotein cholesterol, or alternatively, has a disorder that would benefit from LDL-C reduction , or a combination thereof.

[0014] In a further aspect, the present invention relates to a method for preventing the onset of ASCVD. This method is effective ​​​The process includes administering a certain amount of RNAi agent to a target, wherein the RNAi agent forms a double-stranded region. It is a double-stranded ribonucleic acid containing a nucleus strand and an antisense strand. The antisense strand is the nucleus strand of Sequence ID No. 3. It contains a creotide sequence, and the sense strand contains the nucleotide sequence of SEQ ID NO: 4.

[0015] In another embodiment, the present invention relates to ASCVD, conditions with risks equivalent to ASCVD, and conditions with risks equivalent to CVD. Heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia The patient has the following conditions, requires a reduction in LDL-C, or has a combination of these conditions, and requires treatment. This relates to a method for doing so. This method includes the step of administering an effective amount of RNAi agent to a target, where, RNAi agents are double-stranded ribonucleic acids that include 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. Contains a leotide sequence.

[0016] In certain embodiments, this method treats acute coronary syndrome and / or high LDL-C levels. It is for that purpose.

[0017] In some embodiments, the target is a base higher than approximately 70 mg / dl, for example, approximately 100 mg / dl. It may have a line LDL-C.

[0018] In certain embodiments, the subject is administered RNAi agents more than once. For example, the subject is RNAi The i-agent may be administered multiple times. In some embodiments, the subject is, for example, about once a week. Approximately once every two weeks, approximately once a month, approximately once every two months, approximately once every three months, approximately once every four months, approximately once every six months RNAi agents are administered at regular intervals, such as once a year.

[0019] In a particular embodiment, the method includes the step of evaluating the subject before administering the RNAi agent. This involves measuring one or more biochemical parameters of the subject, including lipid parameters. May include. Examples of parameters that may be measured before administration of RNAi agents include LDL-C, high-density lipoprotein Cholesterol (HDL-C), PCSK9, total cholesterol, triglycerides, non-HDL-C, ultra Low-density lipoprotein cholesterol (VLDL-C), apolipoprotein A1 (Apo-A1), apoli Lipoprotein B (Apo-B), lipoprotein (a) (Lp(a)), C-reactive protein (CRP), glycated hemoglobin Globin A1c, alanine aminotransferase, aspartate aminotransferase Lase, alkaline aminotransferase, creatine kinase, and total bilirubin This includes the level of the RNAi agent. In some embodiments, the evaluation of the subject before administration of the RNAi agent is biochemical. Provides baseline measurements of the latometer. In a particular embodiment, administration of an RNAi agent. The previous assessment focused on the administration of RNAi agents, including the amount and timing of RNAi agent administration. To determine and / or influence.

[0020] In a particular embodiment, the method evaluates the subjects between one or more doses of the RNAi agent. The process includes the step of evaluating one or more biochemical parameters of the subject, including lipid parameters. This may include performing measurements on the target. Measurements may be taken between one or more doses of the RNAi agent. Examples of parameters include LDL-C, HDL-C, PCSK9, total cholesterol, triglycerides, and non-HDL cholesterol. -C, VLDL-C, Apo-A1, Apo-B, Lp(a), CRP, Glycated hemoglobin A1c, Alanine aminotran Spherase, aspartate aminotransferase, alkali aminotransferase This includes levels of enzymes, creatine kinase, and total bilirubin. In some embodiments, The evaluation results then determine the subsequent effects of the RNAi agent, such as the amount of RNAi agent administered and the timing of RNAi agent administration. This may determine and / or influence the dosage (single or multiple doses).

[0021] In some embodiments, administration of RNAi agents compared to baseline LDL-C levels. This reduces LDL-C levels by more than approximately 20%. In a particular embodiment, the baseline A reduction in LDL-C levels of over 20% compared to the previous level occurred 15 days after administration of RNAi agents. The above conditions are maintained for 15 days or more, or for a period of 15 days or more.

[0022] In some embodiments, administration of RNAi agents compared PCSK9 levels to baseline levels. This reduces the PCSK9 level by more than approximately 25%. In a particular embodiment, the baseline A reduction of over 25% in PCSK9 levels compared to the previous level occurred within 30 days after administration of RNAi agents. During the above period, at a point in time of 30 days or more, or maintained over a period of 30 days or more.

[0023] In some embodiments, subjects receive background lipid-lowering therapy such as statins. It may be included. In certain embodiments, the subject continues background lipid-lowering therapy. While doing so, the RNAi agent is administered. In certain embodiments, the subject is maximally tolerable. Patients may be receiving statin therapy. Alternatively, the target group may be those receiving ezetimibe or LDL apheresis. It is acceptable to receive it.

[0024] In other embodiments, the subjects have not received background lipid-lowering therapy.

[0025] In certain embodiments, the subjects do not have active liver disease. For example, the subjects are Alanine aminotransferase and / or aspartic acid exceeding twice the normal upper limit (ULN) Baseline levels of aminotransferase may not be shown. Several implementations Morphologically, subjects do not show baseline levels of total bilirubin greater than 1.5 times the ULN. There are some things that are difficult.

[0026] In another embodiment, the present invention relates to RNAi agents for use in the following methods: (i) (ii) To reduce LDL-C in the subject; (ii) To prevent cardiovascular events in the subject; ( iii) reducing cardiovascular mortality and / or morbidity in the subjects; (iv) ASCV in the subjects To prevent the onset of D; (v) ASCVD, a condition with a risk equivalent to ASCVD, an increased risk of CVD, He has familial hypercholesterolemia that is either homozygous or homozygous. Treatment for patients who need to lower their LDL-C levels, or who have a combination of both. RNAi agents are double-stranded ribonucleotides containing a sense strand and an antisense strand that form a double-stranded region. It is a nucleic acid. The antisense strand contains the nucleotide sequence of sequence number 3, and the sense strand contains sequence number 3. It contains the nucleotide sequence of No. 4. These methods involve administering an effective amount of RNAi agent to a subject. Includes.

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

[0028] [Figure 1] The following shows an RNAi agent according to an embodiment of the present invention. Figure 1 discloses Sequence IDs 4, 3, and 3, respectively, in order of appearance. [Figure 2] A summary of the dosing schedule used in the studies discussed in the examples is provided below. [Figure 3] The average percentage change in LDL-C up to day 360 after a single administration of RNAi agents on day 1 at doses of 200 mg, 300 mg, and 500 mg, according to embodiments of the present invention. [Figure 4] The following shows the time-adjusted average percentage change in LDL-C up to day 360 after a single administration of RNAi agents on day 1 at doses of 200 mg, 300 mg, and 500 mg, and after administration of RNAi agents on day 1 and day 90 at doses of 100 mg, 200 mg, and 300 mg, according to embodiments of the present invention. [Figure 5] The following shows the time-adjusted mean absolute change in LDL-C up to day 360 after a single administration of RNAi agents on day 1 at doses of 200 mg, 300 mg, and 500 mg, and after administration of RNAi agents on day 1 and day 90 at doses of 100 mg, 200 mg, and 300 mg, according to embodiments of the present invention. [Figure 6A] The percentage change in LDL-C between baseline and day 270 in individual patients after a single dose of RNAi agents on day 1 at doses of 200 mg, 300 mg, and 500 mg, according to embodiments of the present invention. [Figure 6B] The percentage change in LDL-C between baseline and day 360 in individual patients after a single dose of RNAi agents on day 1 at 200 mg, 300 mg, and 500 mg doses, according to embodiments of the present invention. [Figure 7] The average percentage change in LDL-C up to day 360 after administration of RNAi agents at doses of 100 mg, 200 mg, and 300 mg according to embodiments of the present invention. [Figure 8] The percentage change in LDL-C between baseline and day 180 in individual patients after administration of an RNAi agent at a dose of 300 mg on day 1 and day 90, according to embodiments of the present invention. [Figure 9A] The percentage change in LDL-C between baseline and day 270 in individual patients after administration of RNAi agents at 100 mg, 200 mg, and 300 mg doses on day 1 and day 90, according to embodiments of the present invention. [Figure 9B] The percentage change in LDL-C between baseline and day 360 in individual patients after administration of RNAi agents at 100 mg, 200 mg, and 300 mg doses on day 1 and day 90, according to embodiments of the present invention. [Figure 10A] Figures 10A to 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 present 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] The graph shows the change in LDL-C from baseline to day 180 for each patient (59 patients) randomly assigned to a two-dose 300-mg RNAi group according to embodiments of the present 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] The graph shows the change in LDL-C from baseline to day 240 for each patient (59 patients) randomly assigned to a two-dose 300-mg RNAi group according to an embodiment of the present 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]The percentage change in LDL-C from baseline to day 90 and day 270 in individual patients after administration of an RNAi agent at a dose of 300 mg on day 1, according to embodiments of the present invention. [Figure 11B] The percentage change in LDL-C from baseline to day 90 and day 270 for individual patients after administration of an RNAi agent at a dose of 300 mg on day 1 and day 90, according to embodiments of the present invention. [Figure 12] The mean percentage change in PCSK9 levels up to day 270 after a single administration of RNAi agents on day 1 at doses of 200 mg, 300 mg, and 500 mg, according to embodiments of the present invention. [Figure 13] The mean percentage change in PCSK9 levels up to day 270 after administration of RNAi agents at doses of 100 mg, 200 mg, and 300 mg, according to embodiments of the present invention. [Figure 14] The following shows modeled results of LDL-C up to 22 months after administration of an RNAi agent administered at a dose of 300 mg twice or three times a year, according to embodiments of the present invention. [Modes for carrying out the invention]

[0029] The present invention, at least in part, is a means of reducing the level 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) within the cell. This allows for the cleavage of the messenger RNA (mRNA) that codes for PCSK9. The fragmented mRNA is degraded and therefore cannot be used for protein translation, and as a result, PSCK9 Protein levels decrease, and as a result, LDL-C levels decrease.

[0030] Therefore, the present invention is a method for lowering LDL-C in a target, wherein the preventive effective dose or This invention provides a method that includes the step of administering an effective amount of RNAi agent, such as a therapeutically effective dose. Furthermore, the method for preventing cardiovascular events in a subject is defined as a prophylactic effective dose or therapeutic effective dose. The present invention provides a method that includes a step of administering an effective amount of RNAi agent to a target, including the quantity. This is a method for reducing cardiovascular mortality and / or morbidity in a subject, and the prophylactic effective dose Alternatively, the present invention provides a method that includes the step of administering an effective amount of RNAi agent, such as a therapeutically effective dose, to a target. Regarding these methods, the target groups are ASCVD, conditions with a risk equivalent to ASCVD, and an increased risk of CVD. heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia They may have, need to lower their LDL-C levels, or have a combination of both.

[0031] Furthermore, the present invention relates to a method for preventing the onset of ASCVD in a subject, wherein the effective preventive dose or The present invention provides a method that includes the step of administering an effective amount of RNAi agent, such as a therapeutically effective dose, to a target.

[0032] In addition, the present invention relates to ASCVD, a state with the same risk as ASCVD, an increased risk of CVD, and heterogeneity. Conjoined familial hypercholesterolemia, homozygous familial hypercholesterolemia, A method for treating subjects who need to lower their LDL-C levels, or who have a combination of both. This method provides an effective dose of RNAi agents, such as a prophylactic or therapeutic dose. This may include the following steps.

[0033] Furthermore, the present invention is for use in the methods described herein, that is, for the target To reduce LDL-C in the subject; to prevent cardiovascular events in the subject To reduce cardiovascular mortality and / or morbidity; to prevent the onset of ASCVD in the target population. To do; and / or ASCVD, a condition with the same risk as ASCVD, an increased risk of CVD, a heterogeneous Homozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia Treating patients who need to lower their LDL-C levels, or who have a combination of both. Regarding RNAi agents for use in [location].

[0034] In some embodiments, cardiovascular events include death, non-fatal myocardial infarction, and severe recurrent aortic treatment. Major ischemia, including but not limited to spontaneous ischemia, stroke, symptomatic pulmonary embolism, and hemorrhage. This could lead to harmful cardiovascular events.

[0035] Methods to lower LDL-C and / or prevent cardiovascular events include having ASCVD, Alternatively, it may be directed at subjects who are in a state of "similar risk to ASCVD." In a particular embodiment, In this context, the subject is considered to have "a condition equivalent to ASCVD in terms of risk" if they have one or more of the following conditions. Possible: A Framingham Risk Score (score > 20%) or a heart condition assessed by an equivalent method. Symptomatic vascular events include subjects with a target LDL-C of less than 100 mg / dl, with a 10-year risk of vascular events. Roam's arteriosclerosis, type 2 diabetes, familial hypercholesterolemia. In some embodiments The subjects may have heterozygous familial hypercholesterolemia.

[0036] Methods to lower LDL-C and / or prevent cardiovascular events also include those related to ASCVD. If you are in a state equivalent to ASCVD in terms of risk, or otherwise have developed heart health problems It may be directed towards individuals exhibiting one or more symptoms or risk factors that indicate a potential for such development. Symptoms / risk factors include having acute coronary syndrome; myocardial infarction, stable or unstable angina. Disease, revascularization of the coronary arteries or other arteries, stroke, transient ischemic attack, or atheroma Having a history of peripheral artery disease of atherosclerotic origin; being male; having heart disease, ASCVD, or This includes having a family history of conditions with a risk level equivalent to ASCVD; having a smoking habit; and being physically active. Not being eligible; having high blood pressure; having high blood cholesterol; having diabetes or prediabetes. Having the condition; being overweight or obese; having a history of pre-eclampsia during pregnancy. and; inability to manage stress and / or anger; being postmenopausal; e.g., salt, saturated fat unhealthy diets, such as those high in trans fats, cholesterol, and / or refined sugars; Being 55 years of age or older; having sleep apnea; being anemic; or a combination of these. This may include cases with a family history of heart disease. or it may be directed at subjects who smoke but do not have high blood pressure. In certain embodiments, this method This may be directed at individuals who have previously experienced a cardiac event.

[0037] In some embodiments, the method of the present invention provides an effective amount, such as a prophylactic effective dose or a therapeutic effective dose. This includes administering RNAi agents to patients with heterozygous familial hypercholesterolemia. This concerns lowering LDL-C and / or preventing cardiovascular events. It is possible. In some embodiments, the method of the present invention may involve a preventive dose or a therapeutic dose, etc. The process includes administering an effective amount of RNAi agent to reduce LDL-C in subjects with ASCVD. It may be about preventing and / or cardiovascular events. In the embodiments, the method of the present invention involves administering an effective amount of RNAi agent, such as a prophylactic or therapeutic effective dose. The process involves reducing LDL-C in subjects with conditions equivalent to ASCVD and those with the same risk. And, and / or it may be about preventing cardiovascular events. In terms of administration, the method of the present invention administers an effective amount of RNAi agent, such as a prophylactic effective dose or a therapeutic effective dose. A process including the following steps to lower LDL-C in subjects with homozygous familial hypercholesterolemia It may be about lowering blood pressure and / or preventing cardiovascular events. In some embodiments, the method of the present invention involves administering an effective amount of RNAi, such as a prophylactic or therapeutic dose. Treatment of a subject with heterozygous familial hypercholesterolemia, including the step of administering a drug. It may be about doing. In some embodiments, the method of the present invention is predetermined ASCVD includes a step of administering an effective amount of RNAi agent, such as a preventive or therapeutically effective dose. It may be about treating the subject. In some embodiments, the method of the present invention This includes a step of administering an effective amount of RNAi agent, such as a prophylactic effective dose or a therapeutic effective dose, to ASCVD and RI The study may be about treating subjects with equivalent conditions. In terms of administration, the method of the present invention administers an effective amount of RNAi agent, such as a prophylactic effective dose or a therapeutic effective dose. The treatment of subjects with homozygous familial hypercholesterolemia includes the following steps. It may be such that the RNAi agent forms a sense that forms a double-stranded region. It may be a double-stranded ribonucleic acid comprising a ribonucleic acid strand and an antisense strand, where the antisense strand is sequence number The nucleotide sequence contains sequence number 3, and the sense strand contains sequence number 4. , fat They may also be on a diet designed to improve quality levels, and / or statins, You may be receiving LDL-lowering therapy such as ezetimibe or LDL apheresis. In that embodiment, the subject may be receiving the maximum permissible statin therapy. Furthermore, the subjects may require a further reduction in LDL-C levels.

[0038] In a particular embodiment, administration of an RNAi agent resulted in a reduction of approximately 2 LDL-C levels compared to baseline. More than 0%, or more than approximately 25%, or more than approximately 30%, or more than approximately 35%, or more than approximately 40% More than approximately 45%, more than approximately 50%, more than approximately 55%, or more than approximately 60% Furthermore, it can reduce the level of LDL-C.

[0039] In some embodiments, the reduction in LDL-C levels after administration of an RNAi agent is due to RNAi Approximately 15 days or more, or approximately 20 days or more, or approximately 30 days or more, or 40 days or more, or approximately 50 days after administration of the drug. More than 60 days, or more than 70 days, or more than 80 days, or more than 90 days, or more than 100 More than a day, or approximately 110 days or more, or approximately 120 days or more, or approximately 130 days or more, or approximately 140 days or more, Approximately 150 days or more, or approximately 160 days or more, or approximately 170 days or more, or approximately 180 days or more, or approximately 190 days or more , or approximately 200 days or more, or approximately 210 days or more, or approximately 220 days or more, or approximately 230 days or more, or approximately 240 More than a day, or approximately 250 days or more, or approximately 260 days or more, or approximately 270 days or more, or approximately 280 days or more, Approximately 290 days or more, or approximately 300 days or more, or approximately 310 days or more, or approximately 320 days or more, or approximately 330 days or more , or for approximately 340 days or more, or approximately 350 days or more, or approximately 360 days or more, at the time of these number of days, Alternatively, it may be maintained over these number of days.

[0040] Administration of RNAi agents at a dose of approximately 100 mg reduces baseline LDL-C levels by day 15. Compared to other methods, LDL-C levels can be reduced by more than 15%, or more than 20%. The reduction lasts for approximately 30 days, or approximately 60 days, or approximately 90 days, or longer. It can be maintained at a specific point or over these days. In some embodiments, The reduction in LDL-C levels compared to line-based LDL-C levels may be over 40% by day 30. For approximately 60 days, approximately 90 days, or longer, at the time of these days, or during these days It can be maintained.

[0041] Administration of RNAi agents at a dose of approximately 200 mg reduces baseline LDL-C levels by day 15. Compared to approximately 15%, or approximately 20%, or approximately 25%, or approximately 30% This can reduce LDL-C levels. This reduction in LDL-C takes approximately 30 days, or approximately 60 days, or approximately 90 days. , or approximately 120 days, or approximately 150 days, or approximately 180 days, or approximately 210 days, or approximately 240 days, or approximately 270 days, Or for a period of approximately 300 days, or approximately 330 days, or approximately 360 days, or longer, during these periods of time It can be maintained at a point or over these days. In some embodiments, the base line The reduction in LDL-C compared to the previous LDL-C level can be over 40% at day 30, and approximately 9 at day 60. For 0 days or longer, at the time of these days, or maintained over these days. It is possible. In certain embodiments, the reduction of LDL-C compared to the baseline LDL-C level is The probability is over 30% on day 15, and approximately 30 days, approximately 60 days, approximately 90 days, approximately 120 days, approximately 150 days, or It may be maintained for a longer period of time, at any point in time during those days, or over those days. In some embodiments, the reduction in LDL-C compared to baseline LDL-C levels occurred on day 180. In this case, or up to day 180, it may be more than 5%. In a particular embodiment, baseline L The reduction in LDL-C levels compared to DL-C levels may be over 20% at day 15, and approximately 30 days, or approximately 60 days, or 90 days, or approximately 120 days, or approximately 150 days, or approximately 180 days, or approximately 210 days, or approximately 240 days , or during a period of approximately 270 days, or approximately 300 days, or approximately 330 days, or approximately 360 days, at the time of these number of days, Or it may be maintained over these days. In certain embodiments, baseline LDL- The reduction in LDL-C compared to the C level was over 25% at day 180, or over 25% at day 270. The probability can exceed 30% on day 360.

[0042] Administration of approximately 300 mg of RNAi agents, by day 15, compared to baseline LDL-C levels. And, the LDL-C level is approximately 15% or more, or approximately 20% or more, or approximately 25% or more, or approximately 30% or more, or approximately 40% or more. It can be reduced. This reduction in LDL-C takes about 30 days, or about 60 days, or about 90 days, or about 120 days, or It is approximately 150 days, or approximately 180 days, or approximately 210 days, or approximately 240 days, or approximately 270 days, or approximately 300 days, or For approximately 330 days, or approximately 360 days, or more, at the time of these days, or these It can be maintained over several days. In some embodiments, it is compared to the baseline LDL-C level. The reduction in LDL-C compared to other conditions may exceed 45% by day 30, and continue for approximately 60, 90, or longer. It may be maintained for a certain number of days, at a certain point in time during those days, or over those days. In this embodiment, the reduction in LDL-C compared to the baseline LDL-C level was approximately on day 15. This can be over 40%, and for approximately 30 days, 60 days, 90 days, 120 days, 150 days, or longer. These may be maintained at or over these number of days. Therefore, the reduction in LDL-C levels compared to baseline levels was over 35% on day 15. It could be approximately 30 days, or approximately 60 days, or approximately 60 days, or approximately 90 days, or approximately 120 days, or approximately 150 days. Or for approximately 180 days, or approximately 210 days, or longer, at the time of these days, or these It can be maintained over a number of days. In some embodiments, the baseline LDL-C level and The comparative reduction in LDL-C may be over 30% at day 15, and may continue for approximately 30 days, or approximately 60 days, or approximately 90 days, or approximately 120 days, or approximately 150 days, or approximately 180 days, or approximately 210 days, or approximately 240 days, or approximately 270 days For days, or approximately 300 days, or approximately 330 days, or approximately 360 days, or for a longer period of time, these days It may be maintained at that point in time or over those days. In a particular embodiment, the base The reduction in LDL-C compared to the in-LDL-C level was over 35% at day 180, or at day 270. The percentage may be over 30% at one point, or over 30% at 360 days.

[0043] Administration of approximately 500 mg of RNAi agents, by day 15, compared to baseline LDL-C levels. And, the LDL-C level is approximately 15% or more, or approximately 20% or more, or approximately 25% or more, or approximately 30% or more, or approximately 40% or more. It can be reduced. This reduction in LDL-C takes about 30 days, or about 60 days, or about 90 days, or about 120 days, or It is approximately 150 days, or approximately 180 days, or approximately 210 days, or approximately 240 days, or approximately 270 days, or approximately 300 days, or For approximately 330 days, or approximately 360 days, or more, at the time of these days, or these It can be maintained over several days. In some embodiments, it is compared to the baseline LDL-C level. The reduction in LDL-C compared to other conditions may be over 50% at day 30, and may continue for approximately 60, 90, or longer. It may be maintained for a certain number of days, at a certain point in time during those days, or over those days. In this embodiment, the reduction in LDL-C compared to the baseline LDL-C level was approximately on day 15. There is a greater than 40% chance of it lasting approximately 30, 60, 90, 120, 150, 180 days, or longer. It can be maintained for a number of days, at the time of these days, or over these days. In this embodiment, the reduction in LDL-C compared to the baseline LDL-C level was approximately 35% on day 15. It could be over % and could be approximately 30 days, or approximately 60 days, or approximately 90 days, or approximately 120 days, or approximately 150 days, or approximately 180 days For days, or approximately 210 days, or approximately 240 days, or longer, at the time of these days, or These can be maintained over the course of several days. In some embodiments, the reduction of LDL-C is based Compared to the line LDL-C level, it may be over 30% at day 15, and for approximately 30 or 60 days. Or approximately 90 days, or approximately 120 days, or approximately 150 days, or approximately 180 days, or approximately 210 days, or approximately 240 days, or For approximately 270 days, or approximately 300 days, or longer, at the time of these days, or these It can be maintained over several days. In certain embodiments, it is compared to the baseline LDL-C level. The reduction in LDL-C observed was approximately 40% or more at day 180, or approximately 30% or more at day 270, or 360 It can be over 30% on day [number].

[0044] Administration of the second dose of RNAi at day 90 resulted in a reduction in LD compared to baseline LDL-C levels. The level of LC can be further reduced. For example, administration of an RNAi agent at a dose of approximately 100 mg on day 1. Subsequently, administration of approximately 100 mg of RNAi agent on day 90 compared to baseline LDL-C levels. Approximately 40% or more between day 104 and day 120 and / or day 150; day 104 to day 120, day 150 Approximately 35% or more by day 180; from day 104 to day 120, day 150, day 180 and / or Approximately 30% more than 210 days; from day 104 to day 120, day 150, day 180, day 210, and day 240 and / or more than approximately 25% by day 270; from day 104 to day 120, day 150, day 180, day 210 Approximately 20% or more by day 240, day 270, day 300 and / or day 330; and / or 104 days Days 120, 150, 180, 210, 240, 270, 300, 330 and / or it may be possible to reduce LDL-C levels by more than 10% by 360 days. In a particular embodiment... Compared to baseline LDL-C levels, the reduction in LDL-C was over 35% at day 180, and This could be over 25% on day 270, or over 10% on day 360.

[0045] After administration of approximately 200 mg of RNAi agent on day 1, approximately 200 mg of RNAi agent on day 90 The administration is performed from day 104 to day 120 and / or day 150, compared to baseline LDL-C levels. Approximately 45% by day 104, 120, 150, 180, 210 and / or 240 days Approximately 40% by the time of the first day; from day 104 to day 120, day 150, day 180, day 210, day 240, day 270 and / or more than approximately 35% by day 300; and / or from day 104 to day 120, day 150, day 180 Approximately 30% by day 1, day 210, day 240, day 270, day 300, day 330 and / or day 360 It is possible to reduce LDL-C levels significantly. In certain embodiments, the baseline LDL-C level and In comparison, the reduction in LDL-C was over 40% at day 180, or over 35% at day 270. This could be over 30% on day 360.

[0046] After administration of approximately 300 mg of RNAi agent on day 1, approximately 300 mg of RNAi agent on day 90 Administration is performed on days 104, 120, 150, and 180, compared to baseline LDL-C levels. and / or more than 50% by day 210; from day 104 to day 120, day 150, day 180, day 210 Approximately 40% by day 240, day 270 and / or day 300; from day 104 to day 120, day 150 Approximately 35% or more by day 180, day 210, day 240, day 270, day 300 and / or day 310; And / or, from day 104 to day 120, day 150, day 180, day 210, day 240, day 270, 30 By day 0, day 330, and / or day 360, LDL-C levels can be reduced by more than approximately 30%. In a particular embodiment, the reduction in LDL-C compared to the baseline LDL-C level occurred on day 180. It could be over 50% at day 1, over 40% at day 270, or over 30% at day 360. ru.

[0047] In a particular embodiment, administration of an RNAi agent compared to baseline PCSK9 levels, More than approximately 25%, or more than approximately 30%, or more than approximately 35%, or more than approximately 40%, or approximately 45% More than % or more than approximately 50% or more than approximately 55% or more than approximately 60% or more than approximately 65% More than approximately 50%, more than approximately 70%, more than approximately 75%, or more than approximately 80% Reduce PCSK9 levels by more than 85%, more than 90%, or more than 95%. It is possible.

[0048] In some embodiments, the reduction in PCSK9 levels occurred approximately 15 days or more after administration of the RNAi agent, Approximately 20 days or more, or approximately 30 days or more, or approximately 40 days or more, or approximately 50 days or more, or approximately 60 days or more, Approximately 70 days or more, or approximately 80 days or more, or approximately 90 days or more, or approximately 100 days or more, or approximately 110 days or more, Or approximately 120 days or more, or approximately 130 days or more, or approximately 140 days or more, or approximately 150 days or more, or approximately 160 days More than or approximately 170 days or more, or approximately 180 days or more, or approximately 190 days or more, or approximately 200 days or more, or approximately 210 days or more, or approximately 220 days or more, or approximately 230 days or more, or approximately 240 days or more, or approximately 250 days or more, Or approximately 260 days or more, or approximately 270 days or more, or approximately 280 days or more, or approximately 290 days or more, or approximately 300 days More than or approximately 310 days or more, approximately 320 days or more, approximately 330 days or more, approximately 340 days or more, or approximately For more than 350 days, or for approximately 360 days or more, at any point in time or over such a period of time. It can be maintained.

[0049] Administration of approximately 100 mg of RNAi agents was compared to baseline PCSK9 levels by day 15. And, more than approximately 20%, or more than approximately 25%, or more than approximately 35%, or more than approximately 40%, PC This can reduce the SK9 level. This reduction in PCSK9 lasts for approximately 30 days, or approximately 60 days, or approximately 90 days. , or for a longer period of time, at the time of these days, or maintained over these days obtain.

[0050] Administration of approximately 200 mg of RNAi agents was compared to baseline PCSK9 levels by day 15. And, more than approximately 20%, or more than approximately 25%, or more than approximately 30%, or more than approximately 35%, This can reduce the PCSK9 level by more than approximately 40%, more than approximately 45%, or more than approximately 50%. This reduction in PCSK9 lasts for approximately 30 days, or approximately 60 days, or approximately 90 days, or longer. It can be maintained over time. In some embodiments, compared to the baseline PCSK9 level Therefore, the reduction in PCSK9 can be over 60% by day 30, and can continue for approximately 60 days, 90 days, or longer. It may be maintained for a number of days, at the time of these days, or over these days. In this embodiment, the reduction in PCSK9 level compared to the baseline PCSK9 level was approximately 3 by day 300. It could be more than 0%.

[0051] Administration of approximately 300 mg of RNAi agents was compared to baseline PCSK9 levels by day 15. And, more than approximately 20%, or more than approximately 25%, or more than approximately 30%, or more than approximately 35%, more than approximately 40%, or more than approximately 45%, or more than approximately 50%, or more than approximately 55%, or approximately It is possible to reduce PCSK9 levels by more than 60%. This reduction in PCSK9 can last for about 30 days, or about 60 days. It can be maintained for days, or approximately 90 days, or longer. In some embodiments Compared to baseline PCSK9 levels, the reduction in PCSK9 was over 60% at day 30. It is possible, for about 60 days, about 90 days, or longer, at the time of these days, or this It can be maintained over the course of several days. In a particular embodiment, the baseline PCSK9 level In comparison, the reduction in PCSK9 can be over 30% up to day 300.

[0052] Administration of approximately 500 mg of RNAi agents was compared to baseline PCSK9 levels by day 15. And, more than approximately 20%, or more than approximately 25%, or more than approximately 30%, or more than approximately 35%, more than approximately 40%, or more than approximately 45%, or more than approximately 50%, or more than approximately 55%, or approximately It is possible to reduce PCSK9 levels by more than 60%. This reduction in PCSK9 can last for about 30 days, or about 60 days. It can be maintained for days, or approximately 90 days, or longer. In some embodiments Compared to baseline PCSK9 levels, the reduction in PCSK9 was over 70% at day 30. It is possible, for about 60 days, about 90 days, or longer, at the time of these days, or these It can be maintained over several days. In some embodiments, the baseline PCSK9 level and In comparison, the reduction in PCSK9 can exceed 30% by day 300.

[0053] Administration of approximately 100 mg of RNAi agent on day 1, followed by an administration of approximately 100 mg of RNAi agent on day 90. The administration of this drug resulted in an increase of approximately 20% compared to baseline PCSK9 levels by day 104, and more than approximately 25%, or more than approximately 30%, or more than approximately 35%, or more than approximately 40%, PCSK9 This can reduce the level of PCSK9. This reduction in PCSK9 can last for about 120 days, or at the time of about 120 days, or at about 1 It can be maintained for 20 days. In some embodiments, compared to the baseline PCSK9 level. In comparison, the reduction in PCSK9 could be over 30% at day 104, and over approximately 150 days. It can be maintained at a single point or for approximately 150 days.

[0054] Approximately 200 mg of RNAi agent administration on day 1, followed by approximately 200 mg of RNAi agent on day 90. The administration of this drug resulted in an increase of approximately 20% compared to baseline PCSK9 levels by day 104, and more than approximately 25%, or more than approximately 30%, or more than approximately 35%, or more than approximately 40%, or approximately It is possible to reduce PCSK9 levels by more than 45%, or approximately more than 50%. This reduction in PCSK9 is It may be maintained for approximately 120 days, at the point of approximately 120 days, or over approximately 120 days. Several implementations Morphologically, the reduction in PCSK9 level compared to baseline was approximately 4 at day 104. It may be greater than 0% and may be maintained for approximately 150 days, at the point of approximately 150 days, or over approximately 150 days.

[0055] Administration of approximately 300 mg of RNAi agent on day 1, followed by an administration of approximately 300 mg of RNAi agent on day 90. The administration of this drug resulted in an increase of approximately 20% compared to baseline PCSK9 levels by day 104, and more than approximately 25%, or more than approximately 30%, or more than approximately 35%, or more than approximately 40%, or approximately It is possible to reduce PCSK9 levels by more than 45%, or approximately more than 50%. This reduction in PCSK9 is Maintained for approximately 120 days, at the point of approximately 120 days, over approximately 120 days, or over 150 days. It is possible. In some embodiments, the PCSK9 level is lower compared to the baseline PCSK9 level. The decrease may be more than 50% or more than 55% on day 120, and over the course of approximately 150 days, at approximately day 150, Alternatively, it can be maintained for approximately 150 days.

[0056] Administration of approximately 100 mg of RNAi agents reduces baseline total cholesterol by day 90. Compared to the level, approximately 5% more, or approximately 10% more, or approximately 15% more, It reduces total cholesterol levels by more than 20%; by day 90, baseline Compared to non-HDL-C levels, it is approximately 5% or more, or approximately 10% or more, or approximately 15% More than approximately 20%, more than approximately 25%, or more than approximately 30%, non-H Reduced DL-C levels; by day 90, approximately 5% compared to baseline Apo-B levels. More than 10%, or more than 15%, or more than 20%, Reduce Apo-B levels by more than approximately 25%; and / or, by day 90, baseline levels Compared to the Lp(a) level, it is about 5% more, or about 10% more, or about 15% more This can reduce the level of Lp(a) by a significant margin.

[0057] Administration of approximately 200 mg of RNAi agents reduces baseline total cholesterol by day 90. Compared to the level, approximately 5% more, or approximately 10% more, or approximately 15% more, Reduce total cholesterol levels by more than 20% or more than 25%; 90 By day [number], levels were approximately 5% higher or 10% higher compared to baseline non-HDL-C levels. More than 15%, or more than 20%, or more than 25%, Reduce non-HDL-C levels by more than 30% or more than 35%; up to day 90 Compared to baseline Apo-B levels, this is approximately 5% or 10% higher, and young Or more than approximately 15%, or more than approximately 20%, or more than approximately 25%, or approximately 30% To reduce Apo-B levels more significantly; and / or, by day 90, baseline Lp(a) levels Compared to Bell, it is about 5% more, or about 10% more, or about 15% more, if This can reduce the Lp(a) level by more than approximately 20%. In some embodiments, on day 180 In this study, total cholesterol was reduced by over 15%, non-HDL-C by over 25%, and Apo-B by approximately 20%. It's incredible.

[0058] Administration of approximately 300 mg of RNAi agents reduces baseline total cholesterol by day 90. Compared to the level, approximately 5% more, or approximately 10% more, or approximately 15% more, Reduce total cholesterol levels by more than 20% or more than 25%; 90 By day [number], levels were approximately 5% higher or 10% higher compared to baseline non-HDL-C levels. More than 15%, or more than 20%, or more than 25%, Or more than approximately 30%, or more than approximately 35%, or more than approximately 40% of non-HDL-C levels It reduces the risk; by day 90, compared to baseline Apo-B levels, it is approximately 5% higher and younger. Or more than approximately 10%, or more than approximately 15%, or more than approximately 20%, or approximately 25% Reduce Apo-B levels by more than 30%, or by more than 35%; And / or, by day 90, the Lp(a) level is approximately 5% higher than the baseline level, if The Lp(a) level is approximately 10% or more, or approximately 15% or more, or approximately 20% or more. It can be reduced.

[0059] Administration of approximately 500 mg of RNAi agents reduces baseline total cholesterol by day 90. Compared to the level, approximately 5% more, or approximately 10% more, or approximately 15% more, Or more than approximately 20%, or more than approximately 25%, or more than approximately 30%, total cholesterol Reduce blood sugar levels; by day 90, blood sugar levels were approximately 5% lower compared to baseline non-HDL-C levels. More than 10%, or more than 15%, or more than 20%, Or more than approximately 25%, or more than approximately 30%, or more than approximately 35%, or approximately 40% Reduce non-HDL-C levels more significantly; by day 90, compare with baseline Apo-B levels. In comparison, more than approximately 5%, or more than approximately 10%, or more than approximately 15%, or approximately 20% More than % or approximately 25% or approximately 30% or approximately 35% Alternatively, reduce the Apo-B level by more than approximately 40%; and / or, by day 90, baseline Compared to the Lp(a) level of IN, it is approximately 5% more, or approximately 10% more, or approximately 15% The level of Lp(a) can be reduced by more, or by more than approximately 20%, in some embodiments. So, at day 180, total cholesterol was reduced by over 25%, and Apo-B was reduced by over 30%. ru.

[0060] Administration of approximately 100 mg of RNAi agent on day 1, followed by an administration of approximately 100 mg of RNAi agent on day 90. The administration of this drug resulted in a reduction of approximately 5% compared to baseline total cholesterol levels by day 180. More than approximately 10%, more than approximately 15%, or more than approximately 20%, total Reduce cholesterol levels; by day 180, compare with baseline non-HDL-C levels. And, more than approximately 5%, or more than approximately 10%, or more than approximately 15%, or approximately 20% Reduce non-HDL-C levels by more, or by more than 25%, or by more than 30%. By day 180, compared to baseline Apo-B levels, there was a difference of approximately 5% or more, or approximately 1%. More than 0%, or more than approximately 15%, or more than approximately 20%, or more than approximately 25% , reduce the Apo-B level; and / or, by day 180, reduce the baseline Lp(a) level and In comparison, the Lp(a) level can be reduced by more than approximately 5%, or more than approximately 10%.

[0061] Approximately 200 mg of RNAi agent administration on day 1, followed by approximately 200 mg of RNAi agent on day 90. The administration of this drug resulted in a reduction of approximately 5% compared to baseline total cholesterol levels by day 180. More than 10%, or more than 15%, or more than 20%, It reduces total cholesterol levels by more than 25%; by day 180, baseline cholesterol levels Compared to non-HDL-C levels, it is approximately 5% or more, or approximately 10% or more, or approximately 15%. More than, or about 20% or about 25% or about 30% Alternatively, reduce non-HDL-C levels by more than approximately 35%; by day 180, baseline Ap Compared to the oB level, approximately 5% more, or approximately 10% more, or approximately 15% more , or more than approximately 20%, or more than approximately 25%, or more than approximately 30%, or Reduce Apo-B levels by more than approximately 35%; and / or reduce baseline L by day 180. Compared to the p(a) level, this is approximately 5% or more, or approximately 10% or more, or approximately 15% This can reduce the level of Lp(a).

[0062] Administration of approximately 300 mg of RNAi agent on day 1, followed by an administration of approximately 300 mg of RNAi agent on day 90. The administration of this drug resulted in a reduction of approximately 5% compared to baseline total cholesterol levels by day 180. More than 10%, or more than 15%, or more than 20%, It reduces total cholesterol levels by more than 25%, or more than 30%; 180 By day [number], levels were approximately 5% higher or 10% higher compared to baseline non-HDL-C levels. More than 15%, or more than 20%, or more than 25%, Or more than approximately 30%, or more than approximately 35%, or more than approximately 40%, or approximately 45% Reduce non-HDL-C levels more significantly; by day 180, compare with baseline Apo-B levels. In comparison, more than approximately 5%, or more than approximately 10%, or more than approximately 15%, or approximately 20% More than % or approximately 25% or approximately 30% or approximately 35% Alternatively, reduce the Apo-B level by more than approximately 40%; and / or, by day 180, base Compared to the line's Lp(a) level, it is approximately 5% or more, or approximately 10% or more. Reduce the Lp(a) level by more than % or more than approximately 20% or more than approximately 25% obtain.

[0063] In a particular embodiment, the method further includes a step of evaluating the subject before administering the RNAi agent. It is possible. The evaluation may be performed on the same day as the administration of the RNAi agent, or one day prior, or two days prior. days prior, or 3 days prior, or 5 days prior, or 6 days prior, or 7 days prior, or 8 days prior, or 9 days prior, or 10 days prior It may be done the day before, or 11 days before, or 12 days before, or 13 days before, or 14 days before. In terms of administration methods, evaluation may be performed over a period of two days or more prior to the administration of the RNAi agent.

[0064] In some embodiments, the evaluation includes age, height, weight, body mass index, race, sex, Whether the subject is receiving any other treatment (e.g., lipid-lowering therapy such as statins), This includes, but is not limited to, the diagnosis of vascular diseases and diabetes, as well as heart rate, blood pressure, and electrocardiogram parameters. This may include measuring one or more physiological parameters or characteristics of the subject without being subjected to any other method. ru.

[0065] In a particular embodiment, the evaluation measures one or more biochemical parameters of the subject. This may include determining the value measured before administration of the RNAi agent. In some embodiments, the value measured before administration of the RNAi agent is "B These can be considered measurements of the "line." 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, glycated hemoglobin A1c, alanine aminotransferase, as Paragate aminotransferase, alkali aminotransferase, creatine These could be, but are not limited to, levels of kinase and total bilirubin. In this embodiment, the evaluation of the subject before administration of the RNAi agent is based on the baseline of biochemical parameters. It may provide measurement values. In a particular embodiment, the evaluation of the subject before administration of the RNAi agent is For example, the amount of RNAi agent, the timing of RNAi agent administration, and other decisions regarding RNAi agent administration and / or This could have an effect.

[0066] In certain embodiments, this method evaluates the subjects between one or more doses of the RNAi agent. This may include evaluation during administration, for example, on days 1, 2, 3, and 4 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, Between 84 and / or 90 days, one or more biochemical parameters of the subject, including lipid parameters, are measured. This may include taking measurements with a meter. Biochemical measurements include LDL-C, HDL-C, PCSK9, Total cholesterol, triglycerides, non-HDL-C, VLDL-C, Apo-A1, apolipoprotein B, Lp(a), CRP, glycated hemoglobin A1c, alanine aminotransferase, asparagine Acid aminotransferase, alkali aminotransferase, creatine kinase This may include, but is not limited to, the levels of total bilirubin, and the total bilirubin level.

[0067] In some embodiments, the evaluation between one or more doses of the RNAi agent is performed using body weight, weight gain, etc. This includes, but is not limited to, a fullness index, heart rate, blood pressure, and electrocardiogram parameters. It is possible to measure physical parameters or characteristics of an object.

[0068] In some embodiments, the evaluation results were used to determine the amount of RNAi agent and the timing of RNAi agent administration. For example, this may determine and / or influence the subsequent administration (one or multiple times) of the RNAi agent. If one or more results from the measurement indicate that the subsequent RNAi agent dose should be reduced by approximately 5%, or approximately 10%, Or approximately 20%, or approximately 30%, or approximately 40%, or approximately 50%, or approximately 60%, or It can be increased or decreased by approximately 70%, 80%, 90%, or more.

[0069] Various biochemical parameters can determine / influence the dose of RNAi agents administered. Example For example, alanine aminotransferase or aspartate aminotransferase that is more than twice the ULN Lanceferase and / or total bilirubin levels exceeding 1.5 times ULN are used in subsequent RNA This can lead to a reduction in the dose of drug i. Another example is the measurement of glycated hemoglobin A1c above 10%. The baseline value may lead to a reduction in subsequent RNAi drug doses. Approximately 1% from the baseline measurement. LDL-C levels that have decreased by more than 5%, less than 10%, or less than 15% are measured in subsequent R This may result in an increase in the dose of NAi agents. The dose may be, for example, about 1%, or about 5%, or about 10%. , or approximately 15%, or approximately 20%, or approximately 25%, or approximately 30%, or approximately 35%, or approximately 40%, or approximately 45%, Alternatively, it may decrease or increase by approximately 50% or more.

[0070] In a particular embodiment, after evaluating the subject before administering the RNAi agent as described above, the subject Lipid-lowering therapy may be administered. Lipid-lowering therapy is a treatment in which the patient receives lipid-lowering therapy regularly. It may be a therapeutic regimen. In some embodiments, the subject is a prior experience with lipid-lowering therapy. Having experience is not required. In a particular embodiment, after the implementation of a lipid-lowering treatment regimen, As mentioned above, the target is one or more biochemical parameters of the target, including lipid parameters. Additional evaluations may be performed to measure the subject. The subject is then described herein. RNAi agents may be administered.

[0071] Administration of RNAi agents The method of the invention is a process of administering an effective amount of RNAi agent to a target, such as a prophylactic effective dose or a therapeutic effective dose. Includes.

[0072] The "effective preventive dose" is the amount used to treat a person who has not yet experienced or exhibited symptoms of the condition but is at risk of contracting it. When administered to a target, it may prevent or improve the symptoms of a certain condition or one or more conditions. It may contain a sufficient amount of RNAi agent to do so. Improving the condition will slow the course of the condition. This includes reducing the severity of conditions that develop later. The "prophylactic effective dose" is determined by the RNAi agent. Method of drug administration, degree of risk of the condition, and medical history, age, weight, family history, genetic makeup, if any. This may vary depending on the type of preceding or concomitant treatment, as well as other individual characteristics of the patient being treated. .

[0073] The "therapeutic effective dose" is the amount administered to the subject when it is effective (for example, in the case of an existing condition or one or more conditions). Sufficient to bring about treatment of the condition (by reducing, improving, or maintaining the symptoms of a number of conditions). This may include the amount of RNAi agent. The "therapeutic effective dose" is determined by the RNAi agent, the method of administration, the degree of risk to the condition, and Medical history, age, weight, family history, genetic makeup, stage of the pathological process, and any preceding or concurrent cases. This may vary depending on the type of treatment used and other individual characteristics of the patient being treated.

[0074] "Baseline" can refer to a state of not receiving treatment, such as before treatment is administered.

[0075] RNAi agents may be administered to the target patient in a fixed dose. A "fixed dose" (e.g., a dose in mg) is... Regardless of any specific subject-related factors such as body weight, the dose used for all subjects. Alternatively, RNAi agents may be administered to the subject in a dose based on body weight (e.g., a dose in mg / kg). This may be done, and this is the dosage of the RNAi agent which changes according to the subject's body weight. In embodiments where dosage is obtained, the RNAi agent is a combination of a fixed dose and a body weight-based dose. It may be administered by hand.

[0076] In a particular embodiment, the RNAi agent is approximately 50 mg to approximately 800 mg, approximately 100 mg to approximately 800 mg, and approximately 150 mg From mg to approximately 800 mg, from approximately 200 mg to approximately 800 mg, from 250 mg to approximately 800 mg, from approximately 300 mg to approximately 800 mg, and from approximately 350 mg. From approximately 800mg, from approximately 400mg to approximately 800mg, from approximately 450mg to approximately 800mg, from approximately 500mg to approximately 800mg, and from approximately 550mg. From approximately 800mg, from approximately 600mg to approximately 800mg, from approximately 650mg to approximately 800mg, from approximately 700mg to approximately 800mg, and from approximately 750mg. From approximately 800mg, from approximately 50mg to approximately 750mg, from approximately 100mg to approximately 750mg, from approximately 150mg to approximately 750mg, and from approximately 200mg. From approximately 750mg, from approximately 250mg to approximately 750mg, from approximately 300mg to approximately 750mg, from approximately 350mg to approximately 750mg, and from approximately 400mg. From approximately 750mg, from approximately 450mg to approximately 750mg, from approximately 500mg to approximately 750mg, from approximately 550mg to approximately 750mg, and from approximately 600mg. From approximately 750mg, from approximately 650mg to approximately 750mg, from approximately 700mg to approximately 750mg, from approximately 50mg to approximately 700mg, and from approximately 100mg. From approximately 700mg, from approximately 150mg to approximately 700mg, from approximately 200mg to approximately 700mg, from approximately 250mg to approximately 700mg, and from approximately 300mg. From approximately 700mg, from approximately 350mg to approximately 700mg, from approximately 400mg to approximately 700mg, from approximately 450mg to approximately 700mg, and from approximately 500mg. From approximately 700mg, from approximately 550mg to approximately 700mg, from approximately 600mg to approximately 700mg, from approximately 650mg to approximately 700mg, and from approximately 50mg. From approximately 650 mg, from approximately 100 mg to approximately 650 mg, from approximately 150 mg to approximately 650 mg, from approximately 200 mg to approximately 650 mg, and from approximately 250 mg. From approximately 650mg, from approximately 300mg to approximately 650mg, from approximately 350mg to approximately 650mg, from approximately 400mg to approximately 650mg, and from approximately 450mg. From approximately 650mg, from approximately 500mg to approximately 650mg, from approximately 550mg to approximately 650mg, from approximately 600mg to approximately 650mg, and approximately 50mg From approximately 600mg, from approximately 100mg to approximately 600mg, from approximately 150mg to approximately 600mg, from approximately 200mg to approximately 600mg, and from approximately 250mg. From approximately 600mg, from approximately 300mg to approximately 600mg, from approximately 350mg to approximately 600mg, from approximately 400mg to approximately 600mg, and from approximately 450mg. From approximately 600mg, from approximately 500mg to approximately 600mg, from approximately 550mg to approximately 600mg, from approximately 50mg to approximately 550mg, and from approximately 100mg. From approximately 550 mg, from approximately 150 mg to approximately 550 mg, from approximately 200 mg to approximately 550 mg, from approximately 250 mg to approximately 550 mg, and from approximately 300 mg. From approximately 550mg, from approximately 350mg to approximately 550mg, from approximately 400mg to approximately 550mg, from approximately 450mg to approximately 550mg, and from approximately 500mg. From approximately 550mg, from approximately 50mg to approximately 500mg, from approximately 100mg to approximately 500mg, from approximately 150mg to approximately 500mg, and from approximately 200mg. From approximately 500mg, from approximately 250mg to approximately 500mg, from approximately 300mg to approximately 500mg, from approximately 350mg to approximately 500mg, and from approximately 400mg. From approximately 500mg, from approximately 450mg to approximately 500mg, from approximately 50mg to approximately 450mg, from approximately 100mg to approximately 450mg, and from approximately 150mg. From approximately 450mg, from approximately 200mg to approximately 450mg, from approximately 250mg to approximately 450mg, from approximately 300mg to approximately 450mg, and from approximately 350mg. From approximately 450 mg, from approximately 400 mg to approximately 450 mg, from approximately 50 mg to approximately 400 mg, from approximately 100 mg to approximately 400 mg, and from approximately 150 mg. From approximately 400mg, from approximately 200mg to approximately 400mg, from approximately 250mg to approximately 400mg, from approximately 300mg to approximately 400mg, and from approximately 350mg. From approximately 400mg, from approximately 50mg to approximately 350mg, from approximately 100mg to approximately 350mg, from approximately 150mg to approximately 350mg, and from approximately 200mg. From approximately 350mg, from approximately 250mg to approximately 350mg, from approximately 300mg to approximately 350mg, from approximately 50mg to approximately 300mg, and from approximately 100mg. From approximately 300 mg, from approximately 150 mg to approximately 300 mg, from approximately 200 mg to approximately 300 mg, or from approximately 250 mg to approximately 300 mg of solids Standard doses, for example, approximately 50 mg, approximately 75 mg, approximately 100 mg, approximately 125 mg, approximately 150 mg, approximately 175 mg, approximately 200 mg, approximately 2 25mg, about 250mg, about 275mg, about 300mg, about 325mg, about 350mg, about 375mg, about 400mg, about 425mg, about 450mg, approximately 475mg, approximately 500mg, approximately 525mg, approximately 550mg, approximately 575mg, approximately 600mg, approximately 625mg, approximately 650mg, The drug is administered to the target group in fixed doses of approximately 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, or 800 mg. The intermediate values ​​and ranges of those listed above are also intended to be part of this invention. It will be done.

[0077] RNAi agents can be administered, for example, as multiple doses repeated at regular intervals. AI agent lasts approximately 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, and 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, approximately 10 months, approximately 11 months, approximately 12 months, approximately 1 year, approximately 13 months, approximately 14 months, approximately 15 months, approximately 16 months Months, approximately 17 months, approximately 18 months, approximately 19 months, approximately 20 months, approximately 21 months, approximately 22 months, approximately 23 months, approximately 24 months It may be administered to the subject at intervals of one month or more, for example, in a chronic administration. In a particular embodiment So, the fixed dose is administered once or multiple times a year, i.e., 2, 3, 4, 5, 6, 7, 8 times. 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd It may be administered to the subject 1, 23, 24 times, or more frequently. In some embodiments, for fixation The dosage is approximately every 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, and 10 weeks. Every, approximately every 11 weeks, approximately every 12 weeks, approximately every 13 weeks, approximately every 14 weeks, approximately every 15 weeks, approximately every 16 weeks, approximately every 17 Weekly, approximately every 18 weeks, approximately every 19 weeks, approximately every 20 weeks, approximately every 22 weeks, approximately every 24 weeks, approximately every 26 weeks, Approximately every 28 weeks, every 30 weeks, every 32 weeks, every 34 weeks, every 36 weeks, every 38 weeks, every 40 weeks Every, approximately every 42 weeks, approximately every 44 weeks, approximately every 46 weeks, approximately every 48 weeks, approximately every 50 weeks, approximately every 52 weeks, or It may be administered to the subject once at longer intervals. In a particular embodiment, the fixed dose is 1 day Once a day, approximately once every two days, approximately once every three days, approximately once every four days, approximately once every five days, approximately once every six days, approximately once every seven days, approximately eight Once a day, once every 9 days, once every 10 days, once every 11 days, once every 12 days, once every 13 days, and once every 14 days. Once every 15 days, once every 16 days, once every 17 days, once every 18 days, once every 19 days, and on the 20th Once a day, once every 30 days, once every 40 days, once every 50 days, once every 60 days, once every 70 days, once every 80 days times, approximately once every 90 days, approximately once every 100 days, approximately once every 110 days, approximately once every 120 days, approximately once every 130 days, approximately every 140 days Once every 150 days, once every 160 days, once every 170 days, once every 180 days, once every 190 days, and once every 2 Once every 00 days, once every approximately 210 days, once every approximately 220 days, once every approximately 230 days, once every approximately 240 days, once every approximately 250 days Approximately once every 260 days, approximately once every 270 days, approximately once every 280 days, approximately once every 290 days, approximately once every 300 days, approximately 310 days Once every 320 days, once every 330 days, once every 340 days, once every 350 days, once every 360 days, and approximately 3 It may be administered to the subject once over a period of 65 days or longer.

[0078] In a particular embodiment, the RNAi agent is administered in a closely spaced "loading phase" followed by This includes a drug regimen that includes a "maintenance phase" in which RNAi agents can be administered at longer intervals. It can be administered as follows: For example, after administration once a week or once every two weeks for one month, administration may be reduced to once a month. It may be repeated for 6 months, 1 year, or longer (e.g., chronic administration).

[0079] In some embodiments, the load phase is the first week, the first two weeks, the first three weeks This may include administering RNAi agents during the first month or so.

[0080] In certain embodiments, one or more doses may be administered during the loading phase. The loading phase consists of the first administration of the RNAi agent on day 1, followed by approximately day 1, day 2, day 3, and day 3. 4th, approximately 5th, approximately 6th, approximately 7th, approximately 8th, approximately 9th, approximately 10th, approximately 11th, approximately 12th, approximately 13th, approximately 14th, Approximately 15 days, approximately 16 days, approximately 17 days, approximately 18 days, approximately 19 days, approximately 20 days, approximately 30 days, approximately 40 days, approximately 50 days, approximately 60 days, Approximately 70 days, approximately 80 days, approximately 90 days, approximately 100 days, approximately 110 days, approximately 120 days, approximately 130 days, approximately 140 days, approximately 150 days, approximately 160 days, approximately 170 days, approximately 180 days, approximately 190 days, approximately 200 days, approximately 210 days, approximately 220 days, approximately 230 days, approximately 240 days, Approximately 250 days, 260 days, 270 days, 280 days, 290 days, 300 days, 310 days, 320 days, 330 days, One or more R after approximately 340 days, 350 days, 360 days, 365 days, or longer This may include the administration of a NAi agent. In some embodiments, the loading phase takes place on day 1. The RNAi agent is administered, followed by approximately 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, and 7 weeks, approximately 8 weeks, approximately 9 weeks, approximately 10 weeks, approximately 13 weeks, approximately 15 weeks, approximately 20 weeks, approximately 25 weeks, approximately 26 Weeks, approximately 30 weeks, approximately 35 weeks, approximately 40 weeks, approximately 45 weeks, approximately 50 weeks, approximately 52 weeks, or longer. This may include a second administration of the RNAi agent after the load. In a particular embodiment, the load The phases involve the administration of RNAi agents on day 1, followed by administration at approximately 1 month, 2 months, 3 months, and 4 months. Months, approximately 5 months, approximately 6 months, approximately 7 months, approximately 8 months, approximately 9 months, approximately 10 months, approximately 11 months, approximately 12 months, Or it may include a second dose of the RNAi agent at a later time. Several embodiments Therefore, the loading phase may include an additional dose of RNAi agent (i.e., more than two doses), The intervals between each dose may vary.

[0081] In a particular embodiment, the RNA agent may be administered on day 1 and again around day 90. .

[0082] In a particular embodiment, the RNAi agent is administered during the loading phase at approximately 50 mg to approximately 800 mg, or approximately 100 mg. From approximately 800mg, from approximately 150mg to approximately 800mg, from approximately 200mg to approximately 800mg, from 250mg to approximately 800mg, from approximately 300mg Approximately 800mg, approximately 350mg to approximately 800mg, approximately 400mg to approximately 800mg, approximately 450mg to approximately 800mg, approximately 500mg to Approximately 800mg, approximately 550mg, approximately 800mg, approximately 600mg to approximately 800mg, approximately 650mg to approximately 800mg, approximately 700mg to approximately 800mg, approximately 750mg to approximately 800mg, approximately 50mg to approximately 750mg, approximately 100mg to approximately 750mg, approximately 150mg to approximately 7 50mg, approximately 200mg to approximately 750mg, approximately 250mg to approximately 750mg, approximately 300mg to approximately 750mg, approximately 350mg to approximately 7 50mg, approximately 400mg to approximately 750mg, approximately 450mg to approximately 750mg, approximately 500mg to approximately 750mg, approximately 550mg to approximately 7 50mg, approximately 600mg to approximately 750mg, approximately 650mg to approximately 750mg, approximately 700mg to approximately 750mg, approximately 50mg to approximately 70 0mg, approximately 100mg to approximately 700mg, approximately 150mg to approximately 700mg, approximately 200mg to approximately 700mg, approximately 250mg to approximately 70 0mg, approximately 300mg to approximately 700mg, approximately 350mg to approximately 700mg, approximately 400mg to approximately 700mg, approximately 450mg to approximately 70 0mg, approximately 500mg to approximately 700mg, approximately 550mg to approximately 700mg, approximately 600mg to approximately 700mg, approximately 650mg to approximately 70 0mg, approximately 50mg to approximately 650mg, approximately 100mg to approximately 650mg, approximately 150mg to approximately 650mg, approximately 200mg to approximately 650mg mg, approximately 250 mg to approximately 650 mg, approximately 300 mg to approximately 650 mg, approximately 350 mg to approximately 650 mg, approximately 400 mg to approximately 650 mg, approximately 450 mg to approximately 650 mg, approximately 500 mg to approximately 650 mg, approximately 550 mg to approximately 650 mg, approximately 600 mg to approximately 650 mg, approximately 50 mg to approximately 600 mg, approximately 100 mg to approximately 600 mg, approximately 150 mg to approximately 600 mg, approximately 200 mg to approximately 600 mg g, approximately 250 mg to approximately 600 mg, approximately 300 mg to approximately 600 mg, approximately 350 mg to approximately 600 mg, approximately 400 mg to approximately 600 mg g, approximately 450 mg to approximately 600 mg, approximately 500 mg to approximately 600 mg, approximately 550 mg to approximately 600 mg, approximately 50 mg to approximately 550 mg Approximately 100mg to 550mg, approximately 150mg to 550mg, approximately 200mg to 550mg, approximately 250mg to 550mg Approximately 300mg to 550mg, approximately 350mg to 550mg, approximately 400mg to 550mg, approximately 450mg to 550mg Approximately 500mg to 550mg, approximately 50mg to 500mg, approximately 100mg to 500mg, approximately 150mg to 500mg Approximately 200mg to 500mg, approximately 250mg to 500mg, approximately 300mg to 500mg, approximately 350mg to 500mg Approximately 400mg to 500mg, approximately 450mg to 500mg, approximately 50mg to 450mg, approximately 100mg to 450mg Approximately 150mg to 450mg, approximately 200mg to 450mg, approximately 250mg to 450mg, approximately 300mg to 450mg Approximately 350mg to 450mg, approximately 400mg to 450mg, approximately 50mg to 400mg, approximately 100mg to 400mg , from about 150 mg to about 400 mg, from about 200 mg to about 400 mg, from about 250 mg to about 400 mg, from about 300 mg to about 400 mg , from about 350 mg to about 400 mg, from about 50 mg to about 350 mg, from about 100 mg to about 350 mg, from about 150 mg to about 350 mg , from about 200 mg to about 350 mg, from about 250 mg to about 350 mg, from about 300 mg to about 350 mg, from about 50 mg to about 300 mg , from about 100 mg to about 300 mg, from about 150 mg to about 300 mg, from about 200 mg to about 300 mg, or from about 250 mg to about 3 00 mg as a fixed dose, for example, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg , about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 m 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 as a fixed dose can be administered to the subject. Intermediate values and ranges of those listed above are also intended to be part of the present invention .

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

[0084] In certain embodiments, the RNAi agent can be administered as a dose of about 300 mg on day 1 and at about day 90 or as a dose of about 300 mg at about day 90.

[0085] In certain embodiments, the maintenance phase involves administration of one or more doses of the RNAi agent to the subject May include. Dosage may be once a month, once every two months, once every three months, once every four months, once every five months, or 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, 18 Once a month, once every 19 months, once every 20 months, once every 21 months, once every 22 months, once every 23 months, once every 24 months Once every 25 months, once every 26 months, once every 27 months, once every 28 months, once every 29 months, once every 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, 43 Once a month, once every 44 months, once every 45 months, once every 46 months, once every 47 months, once every 48 months, or It may be once more frequently. In one particular embodiment, the maintenance dose is once every three months, or every six months. It is administered once a month, once every nine months, or once a year.

[0086] In embodiments of the present invention, the maintenance phase is, for example, repeated multiple times at regular intervals. This may include the administration of RNAi agents in terms of quantity. For example, RNAi agents may be administered for approximately 1 day, 2 days, 3 days, 4 days, Approximately 5 days, approximately 6 days, approximately 1 week, approximately 2 weeks, approximately 3 weeks, approximately 4 weeks, approximately 1 month, approximately 2 months, approximately 3 months, approximately 4 Months, approximately 5 months, approximately 6 months, approximately 7 months, approximately 8 months, approximately 9 months, approximately 10 months, approximately 11 months, approximately 12 months Approximately 1 year, approximately 13 months, approximately 14 months, approximately 15 months, approximately 16 months, approximately 17 months, approximately 18 months, approximately 19 months, At intervals of approximately 20 months, 21 months, 22 months, 23 months, 24 months, or longer, for example It can be administered to the target subject in a chronic manner. In certain embodiments, the fixed dose is once a year or multiple times. Several times, that is, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 11 times, 12 times, 13 times, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 times, or more frequently It can be administered to the subject. In some embodiments, the fixed dose 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, approximately every 11 weeks, approximately every 12 weeks, approximately Every 13 weeks, approximately every 14 weeks, approximately every 15 weeks, approximately every 16 weeks, approximately every 17 weeks, approximately every 18 weeks, approximately every 19 weeks Approximately every 20 weeks, every 22 weeks, every 24 weeks, every 26 weeks, every 28 weeks, every 30 weeks, every 32 weeks Every other week, approximately every 34 weeks, approximately every 36 weeks, approximately every 38 weeks, approximately every 40 weeks, approximately every 42 weeks, approximately every 44 weeks, approximately The drug is administered to the target individual once every 46 weeks, approximately every 48 weeks, approximately every 50 weeks, approximately every 52 weeks, or at longer intervals. It is possible. In certain embodiments, a fixed dose is once a day, about once every two days, about once every three days, about Once every 4 days, approximately once every 5 days, approximately once every 6 days, approximately once every 7 days, approximately once every 8 days, approximately once every 9 days, approximately once every 10 days approximately once every 11 days, approximately once every 12 days, approximately once every 13 days, approximately once every 14 days, approximately once every 15 days, approximately once every 16 days Approximately once every 17 days, approximately once every 18 days, approximately once every 19 days, approximately once every 20 days, approximately once every 30 days, approximately once every 40 days, Approximately once every 50 days, approximately once every 60 days, approximately once every 70 days, approximately once every 80 days, approximately once every 90 days, approximately once every 100 days, Approximately once every 110 days, approximately once every 120 days, approximately once every 130 days, approximately once every 140 days, approximately once every 150 days, approximately once every 160 days times, approximately once every 170 days, approximately once every 180 days, approximately once every 190 days, approximately once every 200 days, approximately once every 210 days, approximately 220 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 about 280 days, once every about 290 days, once every about 300 days, once every about 310 days, once every about 320 days, once every about 330 days, once every about 340 days, once every about 350 days, once every about 360 days, once every about 365 days or longer, it can be administered to a subject. In certain embodiments, the maintenance phase can include administration of a dose of the RNAi agent to the subject every about 3 months, every about 4 months, every about 6 months, every about 9 months, or every about 1 year. The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, 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, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention.

[0087] In certain embodiments, the maintenance phase can include administration of a 300 mg dose of the RNAi agent to the subject every 3 months, every 4 months, every 6 months, every 9 months, or annually. In certain embodiments, the RNAi agent is (1) a loading phase in which the RNAi agent is administered as a 300 mg dose around day 1 and around day 90; and (2) after administration around day 90, as 300 mg every about 6 months.

[0088] The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, 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, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention. [[ID=I6]]The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about I25 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, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention. The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, 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, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention. The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, 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 【75 mg, about 700 mg, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention. The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about I00 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, 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, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention. The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, 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, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention. The maintenance dose(s) can be the same as or different from the loading dose(s). For example, the maintenance dose administered to the subject can be from about 25 mg to about 800 mg, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, 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, 725 mg, about 750 mg, about 775 mg, or about 800 mg. Intermediate values and ranges among those listed above are also intended to be part of the present invention.

[0089] In certain embodiments, the maintenance phase can include administration of a 300 mg dose of the RNAi agent to the subject every 3 months, every 4 months, every 6 months, every 9 months, or annually. In certain embodiments, the RNAi agent is (1) a loading phase in which the RNAi agent is administered as a 300 mg dose around day 1 and around day 90; and (2) after administration around day 90, as 300 mg every about 6 months.

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

[0091] RNAi agents are administered subcutaneously, intravenously, intramuscularly, intraocularly, intrabronchally, intrapleurally, intraperitoneally, intraarterially, and lymphatically. This technology includes, but is not limited to, the internal, intracerebrospinal fluid, and any combination thereof. It can be administered to the subject using any known method of administration in the field. In a preferred embodiment, The medication is administered subcutaneously.

[0092] In some embodiments, administration is via an accumulation injection. The accumulation injection may be a subcutaneous or intramuscular injection. This may include injection. In certain embodiments, the cumulative injection is a subcutaneous injection.

[0093] In some embodiments, administration is via a pump. The pump is either an external pump or surgically implanted. It may be an embedded pump. In a particular embodiment, the pump is an infiltration pump implanted subcutaneously. It is a pressure pump. In other embodiments, the pump is an infusion pump. Infusion pumps are used for intravenous, subcutaneous, intra-arterial, or epidural infusions. In a particular embodiment, the infusion pump can perform subcutaneous infusions. It is a pump. In other embodiments, the pump is surgically implanted to deliver RNAi agents to the liver. It is a pump.

[0094] Other methods of administration include epidural, intracerebral, intraventricular, intranasal, intraarterial, intracardiac, intraosseous, and arachnoid injection. This includes intraluminal, intravitreous, lung, oral, topical, intratracheal, epidermal, or transdermal administration. The mode of administration is: Based on whether local or systemic treatment is preferable, and based on the area to be treated. The administration route and site may be selected to enhance targeting.

[0095] RNAi agents are administered at approximately 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, and 13 minutes. 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 longer It can be administered by intravenous infusion over a period of time such as weekly, bi-weekly, etc. (For example, every two weeks) or regularly for one month, two months, three months, four months, or longer. The treatment may be repeated in between. After the initial treatment regimen, treatment may be administered less frequently. For example, after administering the drug once a week or once every two weeks for three months, the administration may be continued for six months or one year. Or it may be repeated once a month for a longer period of time.

[0096] RNAi agents may be administered in combination with one or more other prophylactic or therapeutic agents. Agent i may be in the same formulation as one or more other prophylactic or therapeutic agents, or RN The RNAi agent may be present in a formulation different from one or more other prophylactic or therapeutic agents. The agent and one or more prophylactic or therapeutic agents may be used simultaneously or in a short period of time with respect to each other, for example. If so, within approximately 1 minute, or within approximately 5 minutes, or within approximately 15 minutes, or within approximately 30 minutes, or within approximately 60 minutes, Approximately 2 hours or less, or approximately 3 hours or less, or approximately 4 hours or less, or approximately 6 hours or less, or approximately 9 hours or less Administer within the same timeframe, or within approximately 12 hours, or within approximately 15 hours, or within approximately 18 hours, or within approximately 24 hours. It is possible.

[0097] One or more other prophylactic or therapeutic agents are used for hypercholesterolemia, atherosclerosis. This includes treatments known to treat lipid disorders such as dyslipidemia or other lipid-related disorders. For example, one or more other preventive or therapeutic agents include HMG-CoA reductase inhibitors, fibrinolytic agents, and fibrous agents. Bile acid chelating agents, niacin, antiplatelet agents, angiotensin-converting enzyme inhibitors, and Geotensin II receptor antagonists, acyl-CoA cholesterol acetyltransferase (AC) AT) inhibitors, cholesterol absorption inhibitors, cholesterol ester transfer proteins CETP inhibitors, microsomal triglyceride transfer protein (MTTP) inhibitors Cholesterol regulators, bile acid regulators, peroxisome proliferation-activating receptor (PPAR) jaw Nist, gene-based therapies, combination vascular protective agents (e.g., AGI-1067 from Atelogenics) , glycoprotein IIb / IIIa inhibitors, aspirin or aspirin-like compounds, IBAT inhibitors, sucrose It may be an allen synthase inhibitor or a monocyte chemotactic protein (MCP)-I inhibitor. For example, one or more other prophylactic or therapeutic agents include, for instance, alirocumab (Plarent). (Company), Evolocumab (Repata), Vococizumab, Roderucizumab, Larpanchizumab, RG7 These could be anti-PCSK9 antibodies such as 652, LY3015014, LPD1462, AX1, ALD1306, or Ig1-PA4.

[0098] Patients receiving RNAi agents The subjects to whom RNAi agents are administered may be humans or non-human animals, and in certain embodiments, the spine These may be vertebrates, and in certain embodiments, mammals. In a preferred embodiment, The elephant is a human. In certain embodiments, the subject is an adult. In some embodiments, The target is the patient.

[0099] The subjects may have a baseline LDL-C level of approximately 70 mg / dl or higher. In a particular embodiment... In this case, the baseline LDL-C level is approximately 80 mg / dl or higher, or approximately 90 mg / dl or higher, or approximately 100 mg / dl / dl or higher, or approximately 110 mg / dl or higher, or approximately 120 mg / dl or higher, or approximately 130 mg / dl or higher, or approximately 140 mg / dl or higher, or approximately 150 mg / dl or higher, or approximately 160 mg / dl or higher, or approximately 170 mg / dl or higher. Or approximately 180 mg / dl or higher, or approximately 190 mg / dl or higher, or approximately 200 mg / dl or higher, or approximately 210 mg / dl or higher, or approximately 220 mg / dl or higher, or approximately 230 mg / dl or higher, or approximately 240 mg / dl or higher, or approximately 250 mg / dl or higher, or approximately 260 mg / dl or higher, or approximately 270 mg / dl or higher, or approximately 280 mg / dl or higher, or approximately 290 mg / dl or higher, or approximately 300 mg It is above / dl.

[0100] In some embodiments, the subject requires a reduction in LDL-C. The target group is those with a blood alcohol level of approximately 50 mg / dl or higher, or approximately 60 mg / dl or higher, or approximately 70 mg / dl or higher, or approximately 80 mg / dl or higher. , or approximately 90 mg / dl or more, or approximately 100 mg / dl or more, or approximately 110 mg / dl or more, or approximately 120 mg / dl or more , or approximately 130 mg / dl or more, or approximately 140 mg / dl or more, or approximately 150 mg / dl or more, or approximately 160 mg / dl or more , or approximately 170 mg / dl or higher, or approximately 180 mg / dl or higher, or approximately 190 mg / dl or higher, or approximately 200 mg / dl or higher , or approximately 210 mg / dl or more, or approximately 220 mg / dl or more, or approximately 230 mg / dl or more, or approximately 240 mg / dl or more , or approximately 250 mg / dl or more, or approximately 260 mg / dl or more, or approximately 270 mg / dl or more, or approximately 280 mg / dl or more They may have LDL-C levels of approximately 290 mg / dl or higher, or approximately 300 mg / dl or higher.

[0101] In some embodiments, the subjects do not have active liver disease. Active liver disease is, Nin aminotransferase, aspartate aminotransferase, and total bil This can be determined by measuring one or more of the biochemical parameters of rubin. Biochemical parameters may be measured at baseline. In certain embodiments, The subjects may have alanine aminotransferase levels less than or equal to 2x (ULN). In some aspects, the target is approximately 1.5 × ULN, or nearly the same as ULN, or slightly less than ULN. It may have nin-aminotransferase levels.

[0102] In some embodiments, the target is aspartate amino acid with a concentration of 2x or less than ULN. It may have a lanceferase level. In certain embodiments, the target is about 1.5 × ULN, or It has an aspartate aminotransferase level that is approximately the same as or less than ULN. obtain.

[0103] In some embodiments, the subject may have a total bilirubin level of 1.5 × ULN or less. In certain aspects, the subjects have total bilirubin levels that are approximately the same as or below ULN. It is possible.

[0104] Alternatively, the subject may have active liver disease. In a particular embodiment, the subject is Approximately 2.5 × ULN, or approximately 3 × ULN, or approximately 3.5 × ULN, or approximately 4 × ULN, or more, such as twice the ULN. It may have alanine aminotransferase levels exceeding (2×). Several implementations In this state, the target is approximately 2.5 × ULN, or approximately 3 × ULN, or approximately 3.5 × ULN, or approximately 4 × ULN, or that It has aspartate aminotransferase levels that are more than twice (2x) those of ULN. Obtain. In certain embodiments, the target is approximately 2 × ULN, approximately 2.5 × ULN, or approximately 3 × ULN, or approximately Total bilirubin levels exceeding 1.5 × ULN, such as 3.5 × ULN, or approximately 4 × ULN, or more. It is possible.

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

[0106] In some embodiments, the subjects may be receiving background lipid-lowering therapy. Even if the subject continues to receive background lipid-lowering therapy while being administered RNAi agents, Alternatively, the subject may discontinue background lipid-lowering therapy. In a particular embodiment In this context, background lipid-lowering therapy can involve statins, and an example of this is atorvasta. Chin, pravastatin, simvastatin, lovastatin, fluvastatin, cerivastatin This includes, but is not limited to, rosuvastatin and pitavastatin. The target group is: You may be receiving the maximum permissible statin therapy; the maximum dose of statins is the maximum dose of this technique. It is publicly known in the field.

[0107] In some embodiments, the subjects are ezetimibe, LDL apheresis, bile acid blockade, You may also be receiving other types of lipid-lowering therapies, such as nicotinic acid and fibrates.

[0108] In other embodiments, the subjects have not received background lipid-lowering therapy.

[0109] In certain embodiments, the subject is designed to improve dietary needs, such as lipid levels. They may be receiving a dietary therapy. Such dietary therapy is publicly known in the art. Yes, for example, a diet designed to improve lipid levels involves eating red meat. And; remove fat from meat; avoid fried foods or high-fat sauces; and avoid eating egg yolks. Ingredients: Skim milk or 1% milk, low-fat frozen yogurt, low-fat ice cream. and use low-fat dairy products such as low-fat cheese; and use fruits and vegetables as sources of fiber. This may include eating certain foods.

[0110] The subjects may have triglyceride levels of approximately 400 mg / dl or less. For example, the subjects may have triglyceride levels of approximately 380 mg / dl. g / dl, or approximately 360 mg / dl, or approximately 340 mg / dl, or approximately 320 mg / dl, or approximately 300 mg / dl, or approximately 280 mg g / dl, or approximately 260 mg / dl, or approximately 240 mg / dl, or approximately 220 mg / dl, or approximately 200 mg / dl, or approximately 180 mg g / dl, or approximately 160 mg / dl, or approximately 140 mg / dl, or approximately 120 mg / dl, or approximately 100 mg / dl, or approximately 80 mg They may have a baseline triglyceride level of / dl or approximately 60 mg / dl.

[0111] In other embodiments, the subject may have a triglyceride level greater than approximately 400 mg / dl. For example, The target group is those with a blood alcohol content of approximately 420 mg / dl, or approximately 440 mg / dl, or approximately 460 mg / dl, or approximately 480 mg / dl, or approximately 500 mg / dl. g / dl, or approximately 520 mg / dl, or approximately 540 mg / dl, or approximately 560 mg / dl, or approximately 580 mg / dl, or approximately 600 mg They may have baseline triglyceride levels of g / dl.

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

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

[0114] In some embodiments, the subjects are those with hyperlipidemia, such as hypercholesterolemia. In a particular embodiment, the subject has heterozygous familial hypercholesterolemia. In another embodiment, the subject has homozygous familial hypercholesterolemia. .

[0115] Alternatively, the target may be eGFR of approximately 25 ml / min, 20 ml / min, or 15 ml / min, or less than 30 ml / min. It may have a full eGFR.

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

[0117] The target group includes individuals with type 2 diabetes, etc., who are untreated or not adequately or completely untreated. It is not necessary to have poorly controlled type 2 diabetes. Diabetes can be identified by a glycated hemoglobin A1c level of at least about 10%. However The target group is less than approximately 10%, for example, about 9%, or about 8%, or about 7%, or about 6%, or about 5%, or It may have a glycated hemoglobin A1c level lower than that.

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

[0119] In some embodiments, glycated hemoglobin A1c levels are measured at baseline.

[0120] The subjects are characterized as Class II, III, or IV by the New York Heart Association (NYHA). Patients do not need to have heart failure. Alternatively, the target group may be the New York Heart Association (NYHA). They may also have heart failure that is more clearly characterized as Class II, III, or IV.

[0121] The target group includes ventricles with approximately 35%, 40%, 45%, 50%, or more than 30%. It may have an ejection rate. In other embodiments, the target is about 25%, or about 20%, or about 15%, or The ventricular ejection fraction may be less than 30%, including cases with a ventricular ejection fraction of less than 30%. It may be measured as a measurement of the ventricular ejection fraction, or as the last known ventricular ejection fraction measured previously. That's fine.

[0122] The target group does not need to experience a major cardiac adverse event within 6 months of receiving RNAi therapy. Cardiac adverse events include death, non-fatal myocardial infarction, severe recurrent ischemia, stroke, and symptomatic events. This includes, but is not limited to, pulmonary embolism and hemorrhage. In some embodiments, the main Cardiac adverse events occurred within approximately 7 months, 8 months, 9 months, and 10 months of RNAi drug administration. Within a month, within approximately 11 months, within approximately 12 months, within approximately 18 months, within approximately 24 months, within approximately 30 months, It has not occurred within approximately 36 months or a longer period, and this includes the occurrence of a major cardiac event. This also includes cases where the organism is not yet alive. In an alternative embodiment, the organism is within approximately 5 months, or approximately 4 months Within a month, or within approximately three months, or within approximately two months, or within approximately one month, or within approximately four weeks, RNAi, including within approximately 3 weeks, or within approximately 2 weeks, or within approximately 1 week, or earlier. A major cardiac event may have occurred within 6 months of administration of the drug.

[0123] The subjects do not necessarily have to have severe hypertension. In some embodiments, severe hypertension High blood pressure is defined as a blood pressure of approximately 190 mmHg, or approximately 200 mmHg, or approximately 220 mmHg, or approximately 240 mmHg, or approximately 260 mmHg. , or systolic blood pressure exceeding approximately 180 mmHg, such as approximately 280 mmHg, approximately 300 mmHg, or more. Systolic blood pressure; and / or approximately 120 mmHg, or approximately 140 mmHg, or approximately 160 mmHg, or approximately 180 mmHg , or a diastolic blood pressure of approximately 200 mmHg, or approximately 220 mmHg, or approximately 240 mmHg, or higher, etc. This can be identified by a diastolic blood pressure exceeding approximately 110 mmHg. In certain embodiments, the target It is not necessary to have uncontrolled severe hypertension. Hypertension is defined as a systolic blood pressure exceeding approximately 180 mmHg and / or approximately 11 mmHg despite antihypertensive therapy. It can be identified by diastolic blood pressure above 0 mmHg. Alternatively, the subjects are those with severe hypertension and / Alternatively, they may have uncontrolled severe hypertension.

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

[0125] The subjects do not need to have a history of any hemorrhagic stroke. Alternatively, the subjects may have a history of hemorrhagic stroke. Even if you have had a stroke.

[0126] The target group is those who have lived for approximately 4 months or less, or approximately 5 months or less, or approximately 6 months or less, or approximately 7 months or less. Within 8 months, or approximately within 9 months, or approximately within 10 months, or approximately within 11 months, or approximately within 12 months or, including a longer period, the patient has not had cardiac arrhythmias within 3 months of administration of the RNAi agent. Alternatively, in another embodiment, the subject has cardiac arrhythmia within 3 months of administration of the RNAi agent. However, cardiac arrhythmias are managed, for example, by medication or surgery. The target group includes those who have had cardiac arrhythmias that have not been controlled by, for example, medication or surgery within the past three months. It's fine if you do that.

[0127] In a particular embodiment, the subject may have one or more of the following: myocardial infarction Stable or unstable angina, revascularization of coronary arteries or other arteries, stroke, transient ischemic attack Having a history of peripheral artery disease of atherosclerosis origin; being male; having heart Having a family history of the disease, ASCVD, or a condition with a risk equivalent to ASCVD; having a smoking habit. and; being physically inactive; having high blood pressure; having high blood cholesterol; diabetes Having illnesses or prediabetic conditions; being overweight or obese; pre-eclampsia during pregnancy. Having a medical history; having uncontrollable stress or anger; being postmenopausal; for example If your diet is 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 those.

[0128] In some embodiments, the target is, for example, Alzheimer's disease, dementia, memory loss. And so on, they may have cognitive impairment. In some embodiments, the subject may have any cognitive impairment It is not harmful.

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

[0130] In embodiments of the present invention, at least one nucleotide on the antisense strand, At least one nucleotide on the sense strand, or both the antisense strand and the sense strand. At least one nucleotide is a modified nucleotide. In some embodiments, , virtually all nucleotides of the antisense strand, virtually all nucleotides of the sense strand If substantially all nucleotides in both the antisense and sense strands are modified nucleotides It is a rheotide. In a particular embodiment, the entire nucleotide of the antisense strand, sense strand All nucleotides, or all nucleotides in both the antisense and sense strands, are modified nucleotides. It is creotide.

[0131] Modifications to nucleotides are performed using techniques and methods known in the art. Obtained. The qualification is obtained from PCT application No. PCT / US2016 / 048666 filed on August 25, 2016 and June 2015. This may include what is described in U.S. application 14 / 650,128, filed on the 5th, and these may include, This is incorporated herein by reference.

[0132] In some embodiments, one or more nucleos of an antisense chain or sense chain. The tide may be 2'-O-methyl (2'-OMe) or 2'-fluoro (2'-F) modified. In the embodiment, one or more nucleotides of the antisense strand or sense strand are 3'-5' -They may be connected via phosphodiester linkages.

[0133] In some embodiments, the double-stranded ribonucleic acid includes a ligand. In certain embodiments, The ligand is conjugated to the 3' end of the sense strand of the double-stranded ribonucleic acid. Examples of this include PCT application no. PCT / US2016 / 048666, filed on August 25, 2016, and June 5, 2015. These are described in U.S. Patent Application No. 14 / 650,128, filed on [date], and are referred to herein by reference. It is incorporated into. In a particular embodiment, the ligand is N-acetylgalactosamine It is a (GalNAc) derivative.

[0134] In a particular embodiment, the RNAi agent is 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3'( The antisense strand of the nucleotide sequence of SEQ ID NO: 3) and 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO: 4) is a double-stranded ribonucleic acid containing the sense strand of the nucleotide sequence, where a, g c and u are 2'-O-methyl(2'-OMe)A, G, C, or U; Af, Gf, Cf, or Uf are 2'-fluorinated r is A, G, C or U; dT is 2'-deoxythymidine; s is phosphorothioate linked Yes, double-stranded ribonucleic acid has a covalently bonded tribranched GalNAc ligand.

[0135] In some embodiments, the RNAi agent is as shown in Figure 1.

[0136] RNAi agents can be in "naked" form or "free RNA" form. "Naked" form "State" refers to the absence of a pharmaceutical composition. For example, a naked RNAi agent is, for example, acetic acid. , citric acid, prolamin, carbonic acid, or phosphoric acid, or combinations thereof may be included. It may be in a sharp buffer. In some embodiments, the buffer is phosphate-buffered saline (P BS) The pH and osmotic pressure of the buffer containing the RNAi agent should be such that it is suitable for administration to the target. It can be adjusted accordingly. In certain embodiments, the RNAi agent may be in water for injection.

[0137] Alternatively, the RNAi agent may be a pharmaceutical composition comprising the RNAi agent and one or more pharmaceutically acceptable carriers. It can be formulated into a substance. Pharmaceutical compositions can be formulated based on the delivery method. For example, composition The substance is delivered parenterally, for example, by intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion. It may be formulated for systemic administration by a doctor; or the composition may be administered, for example, by a continuous pump infusion. It may be formulated for direct delivery into the brain parenchyma by any intracerebral injection; or the composition may be manufactured It can be formulated into a drug. Alternatively, the composition may be administered orally; topically (e.g., by a transdermal patch); for example If, inhalation or inhalation of powders or aerosols, including those administered by a sprayer, into the lungs; trachea It can be formulated for internal administration; intranasal administration; epidermal administration; or transdermal administration.

[0138] Compositions and formulations for parenteral, intraparenchymal (intracerebral), intrathecal, intraventricular, or intrahepatic administration are buffered. Agents, diluents, penetration enhancers, carrier compounds, and other pharmaceutically acceptable carriers or excipients The sterilization solution may include, but is not limited to, other suitable additives such as the above. It will not be done. 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 solvents, capsules, gel capsules, sachets, tablets This may include, but is not limited to, mini-tablet formulations. It also includes thickeners, flavoring agents, and diluents. Emulsifiers, dispersants, or binders may also be used. Pharmaceutical compositions and formulations for topical administration are Transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays The pharmaceutical composition of the present invention may include, but is not limited to, liquid and powder formulations. This may include, but is not limited to, liquid formulations, emulsion formulations, and liposome-containing formulations. The products include tablets, capsules, gel capsules, liquid syrups, soft gels, suppositories, and enemas. It can be formulated in various forms, including, but not limited to, intestinal preparations.

[0139] Examples of pharmaceutically acceptable carriers and specific compositions are provided in the PCT application filed on August 25, 2016. As described in PCT / US2016 / 048666 and U.S. application No. 14 / 650,128 filed on June 5, 2015 These are incorporated herein by reference.

[0140] In a particular embodiment, the RNAi agent is administered in doses of approximately 50 mg to approximately 800 mg, and approximately 100 mg to approximately 800 mg. Approximately 150 mg to approximately 800 mg, approximately 200 mg to approximately 800 mg, 250 mg to approximately 800 mg, approximately 300 mg to approximately 800 mg, Approximately 350mg to approximately 800mg, approximately 400mg to approximately 800mg, approximately 450mg to approximately 800mg, approximately 500mg to approximately 800mg, Approximately 550mg to approximately 800mg, approximately 600mg to approximately 800mg, approximately 650mg to approximately 800mg, approximately 700mg to approximately 800mg, Approximately 750 mg to approximately 800 mg, approximately 50 mg to approximately 750 mg, approximately 100 mg to approximately 750 mg, approximately 150 mg to approximately 750 mg, Approximately 200mg to approximately 750mg, approximately 250mg to approximately 750mg, approximately 300mg to approximately 750mg, approximately 350mg to approximately 750mg, Approximately 400mg to approximately 750mg, approximately 450mg to approximately 750mg, approximately 500mg to approximately 750mg, approximately 550mg to approximately 750mg, Approximately 600mg to approximately 750mg, approximately 650mg to approximately 750mg, approximately 700mg to approximately 750mg, approximately 50mg to approximately 700mg, Approximately 100mg to approximately 700mg, approximately 150mg to approximately 700mg, approximately 200mg to approximately 700mg, approximately 250mg to approximately 700mg, Approximately 300mg to approximately 700mg, approximately 350mg to approximately 700mg, approximately 400mg to approximately 700mg, approximately 450mg to approximately 700mg, Approximately 500mg to approximately 700mg, approximately 550mg to approximately 700mg, approximately 600mg to approximately 700mg, approximately 650mg to approximately 700mg, Approximately 50mg to approximately 650mg, approximately 100mg to approximately 650mg, approximately 150mg to approximately 650mg, approximately 200mg to approximately 650mg, Approximately 250mg to approximately 650mg, approximately 300mg to approximately 650mg, approximately 350mg to approximately 650mg, approximately 400mg to approximately 650mg, Approximately 450mg to approximately 650mg, approximately 500mg to approximately 650mg, approximately 550mg to approximately 650mg, approximately 600mg to approximately 650mg, Approximately 50mg to approximately 600mg, approximately 100mg to approximately 600mg, approximately 150mg to approximately 600mg, approximately 200mg to approximately 600mg, Approximately 250mg to approximately 600mg, approximately 300mg to approximately 600mg, approximately 350mg to approximately 600mg, approximately 400mg to approximately 600mg, Approximately 450mg to approximately 600mg, approximately 500mg to approximately 600mg, approximately 550mg to approximately 600mg, approximately 50mg to approximately 550mg, Approximately 100mg to approximately 550mg, approximately 150mg to approximately 550mg, approximately 200mg to approximately 550mg, approximately 250mg to approximately 550mg, Approximately 300mg to approximately 550mg, approximately 350mg to approximately 550mg, approximately 400mg to approximately 550mg, approximately 450mg to approximately 550mg, Approximately 500mg to 550mg, approximately 50mg to 500mg, approximately 100mg to 500mg, approximately 150mg to 500mg, Approximately 200mg to approximately 500mg, approximately 250mg to approximately 500mg, approximately 300mg to approximately 500mg, approximately 350mg to approximately 500mg, Approximately 400mg to approximately 500mg, approximately 450mg to approximately 500mg, approximately 50mg to approximately 450mg, approximately 100mg to approximately 450mg, Approximately 150mg to approximately 450mg, approximately 200mg to approximately 450mg, approximately 250mg to approximately 450mg, approximately 300mg to approximately 450mg, Approximately 350mg to approximately 450mg, approximately 400mg to approximately 450mg, approximately 50mg to approximately 400mg, approximately 100mg to approximately 400mg, Approximately 150mg to approximately 400mg, approximately 200mg to approximately 400mg, approximately 250mg to approximately 400mg, approximately 300mg to approximately 400mg, Approximately 350mg to approximately 400mg, approximately 50mg to approximately 350mg, approximately 100mg to approximately 350mg, approximately 150mg to approximately 350mg, Approximately 200mg to approximately 350mg, approximately 250mg to approximately 350mg, approximately 300mg to approximately 350mg, approximately 50mg to approximately 300mg, Approximately 100mg to approximately 300mg, approximately 150mg to approximately 300mg, approximately 200mg to approximately 300mg, or approximately 250mg to approximately 300mg Fixed doses in mg, for example, approximately 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg g, about 225mg, about 250mg, about 275mg, 300mg, about 325mg, about 350mg, about 375mg, about 400mg, about 425mg Approximately 450mg, 475mg, 500mg, 525mg, 550mg, 575mg, 600mg, 625mg, 650mg g, deliver a fixed dose of approximately 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, or 800 mg. It can be formulated at an appropriate concentration. The intermediate values ​​and ranges of those listed above are also relevant to this development. It is intended to be part of the Ming Dynasty.

[0141] In some embodiments, the RNAi agent is contained in approximately 1.0 ml, 1.1 ml, 1.2 ml, 1.3 ml, and 1.4 ml. Appropriate amounts of pharmaceutical compositions such as approximately 1.5 ml, 1.6 ml, 1.7 ml, 1.8 ml, 1.9 ml, or 2.0 ml. A composition can be formulated at an appropriate concentration so that a certain volume of the composition is administered to the target. For example, In terms of administration, RNAi agents provide a fixed dose of 300 mg of the drug when approximately 1.5 ml of the formulation is administered to the target. Therefore, it is formulated into appropriate pharmaceutical formulations at approximately 200 mg / ml.

[0142] Unless otherwise defined, all technical and scientific terms used herein are defined in this Specified Terms This has the same meaning as that generally understood by those skilled in the art to which the invention pertains. All publications, patent applications, patents, and other references mentioned in the specification, as well as sequence listings. And the drawings are incorporated by reference in their entirety. In case of any inconsistencies, including definitions, This specification shall take precedence. In addition, the materials, methods, and examples are illustrative and not limiting. That is not what I intended. [Examples]

[0143] In patients with high cardiovascular risk and elevated LDL-C, single or multiple subcutaneous injections and A placebo-controlled, double-blind, no-show study to compare the effects of different doses of RNAi agents administered. A controlled experiment was conducted.

[0144] method RNAi agents are nucleotides 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO: 3) The antisense strand of the sequence and the nucleotype of 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO: 4) A double-stranded ribonucleic acid containing a sense strand of a sequence, where a, g, c, and u are 2'-O-methyl( 2'-OMe)A, G, C or U; Af, Gf, Cf or Uf are 2'-fluoroA, G, C or U; dT is 2'-deoxythymidine; s was phosphorothioate linked. Double-stranded ribonucleic acid is It possessed a covalently bound tribranched GalNAc ligand.

[0145] The primary objective is to evaluate the effect of RNAi therapy on LDL-C levels at day 180. The secondary objective was to evaluate the effects of RNAi agents on the following: (i) 90 days (ii) LDL-C levels in the eye; (ii) LDL-C levels at other points in time; (iii) PCSK9 levels over time; ( iv) Other lipids, lipoproteins, apolipoproteins; (v) Pre-identified whole lipid guide (vi) Percentage of patients achieving drainage; (vii) Individual response to different doses; (vii) Different (viii) Duration of lipid-lowering effect of dose; and safety and tolerability profile of RNAi agents. The study also collected / evaluated the effects of RNAi agents on the following: (a) cardiovascular death, non-fatal heart disease. Cardiovascular events such as myocardial infarction, resuscitated cardiac arrest, and non-fatal strokes (ischemic and hemorrhagic). (b) Antidrug antibodies against RNAi agents.

[0146] This study is for individuals with ASCVD or conditions with equivalent risk to ASCVD (e.g., diabetes mellitus and familial hypercholesterolemia). The study included 501 patients with terolemia and elevated LDL-C. Patients were randomized prior to the study. Standing, and included if all of the following selection criteria are met: (a) Male or female patients aged 18 years or older; (b) ASCVD or a condition with a risk equivalent to ASCVD (symptomatic atherosclerosis, type 2 diabetes) Familial hypercholesterolemia, Framingham Risk Score*, or equivalent method are used for assessment. The 10-year risk of cardiovascular events was assessed for patients with a target LDL-C level of less than 100 mg / dl. Medical history; (c) At the time of screening, for ASCVD patients, serum LDL-C ≥ 1.8 mmol / l (≥ 70 mg / For patients with a condition equivalent to or at risk of ASCVD, serum LDL-C ≥ 2.6 mmol / l (≥ 100 mg) / dl); (d) Fasting triglycerides <4.52 mmol / l (<400 mg / dl) at the time of screening; (e) Estimated glomerular filtration rate (eGFR) calculated using standardized local clinical methodologies. Glomerular filtration rate > 30 ml / min; (f) Patients taking statins should be taking the maximum tolerable dose (clinical trial) Judgment of the responsible physician (investigator); (g) Patients receiving lipid-lowering therapy (such as statins and / or ezetimibe) should screen The medication or dosage should have been taken stably for at least 30 days prior to the trial, and the medication or dosage during participation in the trial should be monitored. There are no plans to change; and (h) If you are willing to give informed consent before commencing any procedure related to the test Furthermore, they must be able to do so and be willing to follow all necessary testing procedures. *Framingham Risk Score > 20%

[0147] Patients who meet any of the following exclusion criteria immediately before randomization will not be eligible to participate in the study. Excluded from: (a) This may prevent participation in the clinical trial, and / or if the patient participates in the clinical trial. , those who may be exposed to significant risk (principal investigator [or co-investigator (delegate)]) (As determined by the authorities), uncontrolled or serious illness, or medical or surgical condition; (b) The opinion of the principal investigator (or co-investigator) that may hinder the interpretation of the clinical trial results. A known underlying medical condition or surgical, physical, or medical condition that may be present; (c) New York Heart Association (NYHA) Class II, III, or IV heart failure, or most recently known left ventricular Ejection fraction <30%; (d) Cardiac arrhythmias within 3 months prior to randomization that are not controlled by medication or surgery; (e) A history of any hemorrhagic stroke; (f) Major adverse cardiac events within 6 months prior to randomization; (g) Uncontrolled severe hypertension: Pre-randomization systolic pressure despite antihypertensive therapy. Diastolic blood pressure > 180 mmHg, or diastolic blood pressure > 110 mmHg; (h) Type 2 diabetes that is not adequately controlled, i.e., glycated hemoglobin A1c before randomization ( HbA1c) > 10.0%; (i) any known current infectious, neoplastic, or metabolic condition of the liver, Active liver disease, or confirmed by measurements repeated at intervals of at least one week. Unexplained alanine aminotransferase and aspartate aminotransferase during cleaning An increase of more than twice the ULN of transferase, or an increase of more than 1.5 times the ULN of total bilirubin. rising; (j) A serious comorbidity in which the patient's average life expectancy is shorter than the duration of the study (e.g., acute systemic infection, or other serious illnesses); this is a treated illness that occurred more than 5 years prior to screening. Includes all cancers except basal cell carcinoma; (k) If you are pregnant or breastfeeding, or may be pregnant, and use at least two methods of contraception (oral contraception) Pregnancy pills, barrier devices, approved contraceptive implants, long-term injectable contraceptives, intrauterine devices or Women who do not wish to use tubal ligation**; defined as more than one year since the last menstrual period. If more than two years have passed since menopause, and if under 55 years of age, a negative pregnancy test within 24 hours of randomization is required. A woman, or a woman who has undergone surgical sterilization, is exempt from this exclusion; (l) A method of contraception that is acceptable throughout the entire trial period (i.e., spermicide-containing condoms) Men who do not wish to use it; (m) Known history of alcohol and / or drug abuse; (n) During the longer of 30 days or 5 half-lives, other investigational drugs or devices Treatment by night; (o) Use of other investigational drugs or devices during the study period; (p) Not limited to, but the following may be reasons why the principal investigator would want to conduct the trial. Any conditions that may prevent this: (i) This includes patients who are unable to communicate with or cooperate with the principal investigator. A condition unsuitable for the test; (ii) Protocol requirements, instructions and limitations related to the test, the nature, scope and occurrence of the test Patients who cannot understand the potential consequences (patients whose cooperation is suspected due to drug abuse or alcohol dependence) including (iii) Not likely to comply with the protocol requirements, instructions, and restrictions related to the test (e.g.) For example, an uncooperative attitude, inability to return for follow-up visits, and inability to complete the test. It seems unlikely; (iv) In the opinion of the principal investigator, participation in the trial would increase the patient's risk. Any medical or surgical condition that is suspected to be such; (v) Someone who is directly involved in the administration of the test, or a close relative of such someone, is involved; (vi) any known cognitive impairment (e.g., Alzheimer's disease); and (q) Previous or current treatment with monoclonal antibodies against PCSK9 (screening of 9) (Within 0 days). **Throughout the entire duration of the exam**

[0148] Patients were screened and randomly assigned to one of six RNAi therapy groups or a placebo group. The allocation is stratified by country and by the 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 doses of RNAi agent or placebo on day 90. I received an injection in my eye.

[0149] An overview of the study design is shown in Figure 2. On day 1, all eligible patients were randomized. The patient received the first subcutaneous dose of either the RNAi agent or placebo. The RNAi agent was administered as a single subcutaneous injection (dose: 200 mg, 3 doses). It was administered as either 00 mg or two injections (dose: 500 mg). The second dose was administered. Patients who were randomized received a second injection of either an RNAi agent or a placebo on day 90.

[0150] The placebo was administered as one or two subcutaneous injections of saline solution. The volume of placebo was as follows: The volume of RNAi agents was matched within the dose and injection regimen, but not between injection regimens. For example, the placebo group receiving a 200 mg dose received 1.0 ml of placebo, but the group receiving a 300 mg dose did not. The placebo group received 1.5 ml of placebo.

[0151] The patient's involvement period in the study includes screening, administration of the study drug, and single or multiple injections. The total period, including the progress of the shooting and the follow-up period up to day 210, was approximately 224 days. Additional follow-up studies were conducted. The maximum duration of intervention, if necessary, was 374 days. LDL-C levels were 20% below the baseline level. Excluding patients who did not return within the time limit, the end-of-trial evaluation was performed on day 210; these patients This is evaluated on day 210, and thereafter until day 360, or when LDL-C is within 20% of the baseline level. Follow-up visits were conducted every 30 days until the condition returned (or until one of the conditions occurred first).

[0152] In the single-dose group (one or two injections on day 1), patients followed the following schedule: (a) Screening: Day 14 to Day 1 (b) Randomization and initiation of investigational drug: Day 1 (c) Treatment phase: Day 1 (d) Follow-up survey: (i) Follow-up survey: Day 2 to Day 210; EOS on Day 210 (ii) Additional follow-up (patients whose LDL-C levels did not recover to 80% of baseline levels) In the case of patients; the patient will be re-examined monthly for follow-up until this level is reached, or until day 360. Patients will be required to visit the hospital on the following days: day 240, day 270, day 300, day 330, and day 360.

[0153] In the two-dose group (one injection on day 1 and one on day 90), patients follow the following schedule: tta: (a) Screening: Day 14 to Day 1 (b) Randomization and initiation of investigational drug: Day 1 (c) Treatment phase: Day 1 to Day 90 (d) Follow-up survey: (i) Follow-up survey: Day 91 to Day 210; EOS on Day 210 (ii) Additional follow-up (patients whose LDL-C levels did not recover to 80% of baseline levels) In the case of patients; the patient will be re-examined monthly for follow-up until this level is reached, or until day 360. Patients will be required to visit the hospital on the following days: day 240, day 270, day 300, day 330, and day 360.

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

[0155] The secondary endpoint was (i) the percentage change in LDL-C from baseline to day 90. (ii) From baseline to day 14, day 30, day 60, day 120, day 150, and day 210 Percentage change in LDL-C; (iii) LDL-C levels at days 180 and 210 are below baseline levels. (iv) The proportion of patients in each group with more than 80% LDL; (iv) Treated patients with 80% or more LDL at baseline -The time until the protein returns to C or PCSK9; (v)On days 90, 120, and 180, Treatment reduces LDL-C levels to less than 25 mg / dl, less than 50 mg / dl, less than 70 mg / dl, and less than 100 mg / dl. (vi) Individual response defined as the number of patients who reached the target; (vi) on day 90, day 120, and day 180 (vii) The proportion of patients in each group who have a 50% or greater reduction in LDL-C from baseline; Days 14, 30, 60, 90, 104, 120, 150, and 180 from the line, (viii) Percentage change in PCSK9 level up to day 210; (viii) 210 days from each subsequent visit Up to the eye, other lipids, lipoproteins, and apolipoproteins from baseline Percentage change; and (ix) overall lipid modification target with respect to ASCVD risk level This was evaluated by determining the proportion of patients in each group.

[0156] The effectiveness was evaluated using total cholesterol, triglycerides, HDL-C, non-HDL-C, VLDL-C, and Apo- In relation to the levels of LDL-C lipids and lipoproteins, including A1, Apo-B, Lp(a), CRP, and PCSK9 This included measuring the effectiveness of the RNAi agent.

[0157] Adverse events, serious adverse events, vital signs, clinical laboratory values ​​(blood tests, coagulation tests, chemistry tests) Medical tests (including urine tests) and electrocardiograms (ECG) are collected at specific visits until the end of service (EOS) visit (day 210). They were collected. Between visits for additional monthly follow-up, adverse events, serious adverse events, and Clinical laboratory values ​​were continuously evaluated (LDL-C levels did not recover to above 80% of baseline values). (In the case of patients who have had a heart attack). Cardiovascular events include cardiovascular death, non-fatal myocardial infarction, and major coronary artery injury. Events (death due to CHD, resuscitation-induced cardiac arrest, non-fatal myocardial infarction), ischemic stroke, and Information regarding cardiovascular events such as hemorrhagic stroke is reported as an adverse event. Ta.

[0158] In addition, anti-drug antibodies were evaluated for RNAi agents. Anti-drug antibody formation was observed on day 1 (after injection). (Before and 4 hours later), and on the 30th, 60th, 90th, 120th, 150th, and 180th days (150th day and (Only in patients who received the second dose of RNAi on day 180), and on day 210, The response to anti-drug antibodies was evaluated until it became negative within the study period. LDL-C levels were baseline. For patients whose I-value has not returned to above 80%, when LDL-C returns to the normal limit, or The formation of anti-drug antibodies was evaluated during one of the one-year follow-up visits.

[0159] An independent Data Monitoring Committee (DMC) determined that the first 40 patients were either receiving RNAi therapy or a placebo. Safety data can be examined from the time the patient completes a follow-up visit on the 14th day after receiving the initial injection. They discussed it. After that, DMC reviewed the safety data every two months until the end of the trial.

[0160] The primary endpoint is the least squares mean percentage from baseline to day 180. This was analyzed as a change. This was compared between the test group, baseline values, and follow-up by scheduled visits. The study, including the interaction between the test group and scheduled visits, was conducted using a repeated measures linear effects model. The calculations were performed. The analysis combined treatment at each visit as a fixed effect and patients as a random effect. Using the incorporated autoregressive variance structure, the PROC MIXED procedure is used in SAS software. The study was conducted using six RNAi agents for both the primary and secondary endpoints. Dunnett's test was used to compare the group with the placebo control group, and the p-value was used for multiple comparisons. The dosage strategies, namely single doses and double doses, were adjusted separately. Analysis was performed. The significance level for type I errors in the two-tailed test was 0.05.

[0161] All patients who received at least one dose of RNAi or placebo were included in the safety analysis. The safety population was determined to receive at least one dose of the investigational drug. The patient was administered the drug, and both baseline and 180-day follow-up LDL-C level measurements were available. Furthermore, all patients were defined as randomly assigned. Treatment intention analysis was performed due to insufficient data. For patients who were identified, the attribution was used.

[0162] Time-course data is shown as the mean of the 95% confidence interval. Variation in patient responses is, The data is shown graphically using a waterfall plot. The analysis was performed using SAS software. The test was conducted using version 9.2 or later (SAS Institute).

[0163] result The baseline demographics and medical history shown in Table 1 and Table 2 are based on patient data. This demonstrates that the balance of participants was well maintained. During the trial, 73% of patients were st They were undergoing tin therapy, and 31% were taking ezetimibe.

[0164] A single dose of RNAi on day 1 resulted in a 44% increase in baseline at day 30 across all doses. This resulted in an average reduction in LDL-C levels ranging from 5% to 51.5% (see Figures 3 and 4), with the lowest levels occurring around day 60. The points were shown. As shown in Figure 3, a single dose of 200 mg of RNAi agent was administered on day 30 and day 60. Furthermore, LDL-C levels were reduced by more than 40% from baseline by day 90, and by 25% by day 270. The single dose of RNAi agents at 300 mg and 500 mg doses was administered from day 15 to day 150. It reduced LDL-C by over 40% compared to the baseline, and by 30% by day 270 (Figure 3). For each RNAi drug dose intensity, LDL-C remained more than 20% lower than baseline up to day 360. It was up to this point (Figure 3). The least squares mean reduction was greater for RNAi agents than for placebo (2.1% increase). The effect was significantly greater after a single dose reduction of 27.9-41.9% (P<0.001 for all doses) (Ta (Table 4)). Average LDL-C from baseline across doses of 200 mg, 300 mg, and 500 mg. The average reduction was between 26% and 34.3% at day 270, and between 30.2% and 32.2% at day 360. It was within the range of [a certain range]. Furthermore, the time-adjusted percentage of LDL-C between baseline and day 360 [a certain range]. Changes (i.e., LDL-C levels measured at each time point between baseline and day 360) The mean percentage change in DL-C was -31.6% for doses of 200 mg, 300 mg, and 500 mg, respectively. The values ​​were -38.1% and -39.8% (see Figure 4). LDL-C levels between baseline and day 360 were [value missing]. Inter-adjusted absolute change (i.e., LDL-C measured at each time point between baseline and day 360) The mean absolute change in LDL-C was -39 for doses of 200 mg, 300 mg, and 500 mg, respectively. The values ​​were 1 mg / dl, -43.6 mg / dl, and -53.0 mg / dl (see Figure 5). Looking at the responses of individual patients... The coefficient of variation of the change in LDL-C in individual patients administered 200 mg, 300 mg, and 500 mg was: From the baseline to day 270, the rates were 45.2%, 60.8%, and 50.7%, respectively. From the start to day 360, the rates were 45.1%, 56.8%, and 50.2%, respectively (see Figures 6A and 6B).

[0165] The administration of the second dose of RNAi at day 90 further reduced LDL-C compared to baseline. It was reduced. The mean reduction in LDL-C across all doses was 34.2% to 44.1% at day 90. The intermittent and day 120 rates ranged from 41.1% to 54.6% (see Figure 7). Two-dose regimen The difference between men and placebo was significant: at day 180, patients who received two doses of RNAi were significantly more advanced. The least-squares mean reduction in LDL-C levels from baseline in these individuals ranged from 35.5% to 52.6%. On the other hand, the placebo group showed a 1.8% increase from baseline (compared to placebo). In the comparison, P<0.001 (see Table 4). Second dose of RNAi agents at 100 mg and 200 mg doses. The treatment maintained LDL-C levels below baseline, exceeding 30% and 40%, respectively, until day 210 (Figure 7). (See reference). The second dose of 300 mg was administered from day 120 to day 210, when LDL-C was reduced by more than 50% and the baseline was maintained. The baseline was maintained below the baseline level across 100 mg, 200 mg, and 300 mg doses. The average reduction in LDL-C from day 270 was between 25.6% and 43.4%, and at day 360... The range was between 13.3% (100 mg dose) and 33.3% (300 mg dose). In addition, baseline The time-adjusted percentage change in LDL-C between day 1 and day 360 was observed for 200 mg, 300 mg, and 50 mg. At the 0-mg dose, the percentages were -31.0%, -41.2%, and -46.8%, respectively (see Figure 4). The time-adjusted absolute change in LDL-C between day 60 and day 60 was observed for the 200-mg, 300-mg, and 500-mg doses. The values ​​were -39.6 mg / dl, -39.1 mg / dl, and -57.7 mg / dl, respectively (see Figure 5). The response is shown in Figures 8 and 9. The mean LDL-C from baseline at day 180. 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 was 45.7 from baseline to day 270. The percentages were 82.3% and 73.7%, respectively, from baseline to day 360, which were 46.0% and 77.5%. , and 74.2% (see Figures 9A-9B).

[0166] In patients who received placebo against the background of the maximum possible dose of statins, There was considerable fluctuation in the LDL cholesterol level from the sample at day 180. (Mean [±SD] absolute difference, -0.7±25.6 mg per deciliter [-0.02±0.66 mg per liter) (See Figure 10A). In contrast, all patients who received two doses of 300 mg of RNAi agent showed the same results on day 180. It shows a decrease in LDL-C levels (mean absolute change in LDL cholesterol levels, per deciliter) -64.2±20.7 mg [-1.66±0.54 mmol per liter] (see Figure 10B), 54% of patients had more than 50% A reduction was observed. In this RNAi dose group, 5%, 48%, and 66% of patients experienced a decrease of 1 day at day 180. Less than 25 mg per liter (0.65 mmol per liter), 50 mg per deciliter (1 liter Less than 1.3 mmol per tol, and less than 70 mg per deciliter (1.8 mmol per liter). Each patient had a different LDL-C level. On day 240, the individual cholesterol levels in this patient group were Terol levels remained lower than baseline (see Figure 10C).

[0167] A comparison of individual patient data between a single dose of 300 mg and a double dose of 300 mg is shown below. Administration of 300 mg of RNAi at day 1, day 90, day 270, and every 6 months thereafter within the individual This suggests that it will reduce variability and lead to a sustained reduction of approximately 50% in LDL-C. (See Figures 11A-11B).

[0168] Furthermore, there was a significant reduction in the levels of non-HDL-C and Apo-B, but the high-sensitivity C-reactive protein In patients randomly assigned to receive RNAi therapy, there was a significant change in the level. There was none. Percentage change from baseline for additional lipid measurements is shown in Table This is shown in Tables 3 through 5.

[0169] As shown in Figures 12 and 13, after the RNAi agent is administered on day 1, 100 mg to 500 mg Across the range of RNAi drug doses, PCSK9 levels increased from baseline levels by a mean of 59.6% and 68%. It was reduced by 0.7%. On day 30, PCSK9 levels were 66.2% lower than baseline levels. The percentage further decreased to between 74.0%, and similar decreases occurred on days 60 and 90. RNAi agents In patients who received a single dose, the mean reduction in PCSK9 levels at day 180 was 47.9% to 59%. The range was between 3% (P<0.001 for each dose vs. placebo) (see Table 4). 300 mg Alternatively, in patients who received a single dose of 500 mg, PCSK9 levels increased by 60% from baseline up to day 150. The percentage remained below 1%. In contrast, in patients who received two doses of RNAi, the second Further reductions in PCSK9 levels were observed after dose administration. At day 90, these patients were 4 There was a reduction from 7.0% to 62.8%, and by day 120, there was a reduction from 60.4% to 74.5%. Day 180 In this study, mean PCSK9 levels from baseline in patients who received two doses of RNAi agents The reduction ranged from 53.2% to 69.1% (P<0.001 for each dose vs. placebo) (Table See 4 (Table 4). In patients who received two doses of 200 mg or 300 mg, PCSK9 levels remained elevated until day 240. The levels remained more than 60% lower than baseline. Both single-dose and double-dose RNAi therapy were administered. In relation to the regimen, the reduction in PCSK9 levels at day 270 was greater than approximately 40% (Figure 12 and (See Figure 13).

[0170] To predict LDL-C levels over time when RNAi agents are administered at 300 mg twice or three times a year. The model was developed and validated. The model uses observational data up to day 210 and prediction data. The results were verified with minimal differences between the two. The results at day 270 were followed up to day 210. This was correctly predicted by a pharmacodynamic model using data from the patients who received the treatment. Figure 14 shows 22 This shows the modeled LDL-C levels throughout the month.

[0171] On day 210, 76% of patients who received RNAi agents and 76% of patients who received placebo had Adverse events were reported (see also Table 7 and Table 6). Most of these events (95%) occurred. The severity was mild or moderate (grade 1 or 2). The incidence of serious adverse events was RN The incidence was 11% in patients who received the AI ​​drug and 8% in patients who received the placebo. Two patients experienced adverse events. Two participants discontinued their participation in the clinical trial: one due to a herpes zoster infection (placebo group), and the other due to an influenza virus. The most common adverse events were due to luenza or nasopharyngitis (two doses of 100 mg RNAi agent group). (occurring in more than 2% of patients) include muscle pain, headache, fatigue, nasopharyngitis, lower back pain, high blood pressure, diarrhea, and dizziness. The incidence of these events was significantly different between the group that received RNAi agents and the group that received placebo. It was indeed as expected.

[0172] Injection site reactions occurred in 4% of patients who received a single dose of RNAi agents and 7% of patients who received two doses. Injection site reactions occurred (5% in combined proportions) after one or both doses; compared to placebo. It did not occur in the assigned patients (Table 7; Table 8 and Table 9 also show this). reference).

[0173] Two patients had elevated levels of hepatic aspartate aminotransferase ( (More than 3 times the upper limit of the normal range), one patient in the single-dose placebo group, and one patient in the single-dose 300 mg RNAi group. This was the drug group; patients in this 300-mg RNAi drug group had hepatic alanine aminotransferase There was also an increase in Bell's score. In addition, two more patients (one in the 100 mg twice-dose RNAi group and one in the 3 twice-dose group) One patient in the 00mg RNAi group also had elevated alanine aminotransferase levels. All elevations of aminotransferase were transient. Levels were normal at baseline. In patients with bilirubin levels, bilirubin levels associated with RNAi agents or placebo were observed. There was no increase in rilbin levels, but hepatocyte damage accompanied by jaundice (not cholestatic disorder) was present. If administered with a drug that causes this, the patient is at high risk of fatal drug-induced liver injury. There were no cases that met the definition of Hy's Law. One patient received an antidote before the first injection. The test was positive for physical antibodies; no cases of other anti-drug antibodies have been reported (Table 10). (See Table 10).

[0174] In the follow-up study from day 210 to day 360, no additional safety findings were observed. (See Table 11).

[0175] Two deaths occurred in the later stages of the clinical trial. The first case involved a patient who suffered from long-term vascular complications and frequent angina. This occurred in patients who were randomly assigned to the 500 mg single-dose RNAi group. The first patient presented with cardiac arrest and died on day 104. The second death occurred in a patient who underwent percutaneous regeneration after participating in the study. A male patient in the 2-dose 200-mg RNAi group who had a thoracic aortic aneurysm and subsequently developed a fistula and sepsis. He died on the 198th day of the clinical trial.

[0176] conclusion These results indicate that RNAi agents significantly reduce LDL-C in patients over several months. This showed that RNAi agent administration affected Lp(a) and total cholesterol among other lipid parameters. It reduced and increased HDL-C. RNAi agents were effective against PCSK9, LDL-C, non-HDL-C, and Apo-B. A dose-response effect was observed. Individual patient variability in LDL-C was substantially eliminated, and sparseness was observed. The effects were sustained even during administration.

[0177] In addition, the administration of RNAi agents is well tolerated by patients and is not associated with any major safety issues. It hadn't been done. TEAE is rare, mild to moderate, and the burden of injection was minimal. Ta.

[0178] Overall, RNAi agents offer an 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] While embodiments of the present invention have been described in detail above, it should be understood that the present invention as defined in the above paragraphs should not be limited to the specific details shown above, as many obvious modifications thereto are possible without departing from the spirit or scope of the invention. Various embodiments of the present invention are shown below. 1. A method for lowering low-density lipoprotein cholesterol in a subject, comprising the step of administering a therapeutically effective dose of an interfering ribonucleic acid (RNAi) agent to the subject. RNAi is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 2. A method for preventing cardiac events in a subject, comprising the step of administering a therapeutically effective amount of interfering ribonucleic acid (RNAi) agent to the subject, RNAi is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 3. A method for reducing cardiovascular mortality and morbidity in a subject, comprising the step of administering a therapeutically effective dose of an interfering ribonucleic acid (RNAi) agent to the subject, RNAi is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 4. The method according to any one of items 1 to 3 above, wherein the subject has atherosclerotic cardiovascular disease (ASCVD), a condition with a risk equivalent to ASCVD, an increased risk of cardiovascular disease, heterozygous familial hypercholesterolemia, or homozygous familial hypercholesterolemia, or needs to lower LDL-C, or has a combination thereof. 5. The method according to any one of 1 to 4 above, wherein the subject has a baseline low-density lipoprotein (LDL-C) level of approximately 70 mg / dl or higher. 6. The method described in any of items 1 to 5 above, wherein the subject does not have active liver disease. 7. The method according to any one of items 1 to 6 above, wherein active liver disease is identified by one or more of the following characteristics: alanine aminotransferase (ALT) greater than twice the upper limit of normal (ULN); aspartate aminotransferase (AST) greater than twice the ULN; and total bilirubin greater than 1.5 times the ULN. 8. The method described in any of items 1 to 7 above, wherein the subject has active liver disease. 9. The method according to 8 above, wherein active liver disease is identified by one or more of the following characteristics: alanine aminotransferase (ALT) greater than twice the upper limit of normal (ULN); aspartate aminotransferase (AST) greater than twice the ULN; and total bilirubin greater than 1.5 times the ULN. 10. The method described in any of items 1 to 9 above, wherein the subject is being treated with background lipid-lowering therapy. 11. The method according to 10 above, wherein background lipid-lowering therapy is selected from statins, ezetimibe, and LDL apheresis. 12. The method according to 10 or 11 above, wherein the background lipid-lowering therapy is a statin. 13. Any method described in 10 to 12 above, wherein the subject is receiving the maximum acceptable statin therapy. 14. The method according to any one of items 10 to 13 above, wherein background lipid-lowering therapy is maintained while the subject is receiving RNAi agents. 15. The method described in any of items 1 to 9 above, provided that the subject has not received background lipid-lowering therapy. 16. The method described in any of items 1 to 15 above, wherein the subject has a baseline triglyceride level of approximately 400 mg / dl or less. 17. The method according to any one of 1 to 16 above, wherein the subject has a baseline estimated glomerular filtration rate (eGFR) of at least approximately 30 ml / min. 18. The method described in any of items 1 to 17 above, wherein the subject does not have poorly managed type 2 diabetes. 19. The method according to 18 above, wherein poorly controlled type 2 diabetes is identified by baseline glycated hemoglobin A1c levels of at least about 10%. 20. Any method described in 1 to 19 above, wherein the subject does not have New York Heart Association (NYHA) class II, III, or IV heart failure. 21. The method described in any of items 1 to 20 above, wherein the most recent known ventricular ejection fraction of the subject is 30% or greater. 22. The method described in any of items 1 to 21 above, wherein the subject has not experienced a major adverse cardiac event within 6 months of administration of the RNAi agent. 23. The method described in any of items 1 to 22 above, wherein the subject has not experienced a hemorrhagic stroke. 24. The method according to any one of items 1 to 23 above, wherein the subject does not have cardiac arrhythmias within 3 months of administration of the RNAi agent. 25. The method according to any one of items 1 to 24 above, wherein the subject does not have cardiac arrhythmias within 3 months of administration of RNAi agents and is not controlled by medication or resection. 26. The method described in any of items 1 to 25 above, wherein the subject does not have cardiac arrhythmia. 27. Any method described in 1 to 26 above, wherein the subject is an adult. 28. The method described in any of items 1 to 27 above, wherein the subject has heterozygous familial hypercholesterolemia or homozygous familial hypercholesterolemia. 29. The method described in any of items 1 to 28 above, wherein the subject requires a reduction in LDL-C. 30. The method according to any one of items 1 to 29 above, wherein administration of an RNAi agent reduces the LDL-C level by more than approximately 20% compared to the baseline LDL-C level. 31. The method according to any one of items 1 to 30 above, wherein a reduction in LDL-C levels of more than approximately 20% compared to baseline levels is maintained for 15 days or more after administration of the RNAi agent. 32. The method according to any one of items 1 to 31 above, wherein administration of an RNAi agent reduces the level of PCSK9 by more than approximately 25% compared to the baseline level of PCSK9, and the baseline level of PCSK9 is measured before administration of the RNAi agent. 33. The method according to any one of items 1 to 32 above, wherein a reduction in PCSK9 levels of more than approximately 25% compared to baseline levels is maintained for more than 30 days after administration of the RNAi agent. 34. The method according to any one of 2 or 4 to 33 above, wherein the cardiac event is selected from the group consisting of death, non-fatal myocardial infarction, severe recurrent ischemia, stroke, symptomatic pulmonary embolism, and hemorrhage. 35. The method according to any one of items 1 to 34 above, wherein the administration of the RNAi agent includes a loading phase and a maintenance phase. 36. The method according to 35 above, wherein the loading phase includes administering an RNAi agent as one or more doses. 37. The method according to 36 above, wherein the loading phase includes administering an RNAi agent in at least two doses. 38. The method according to 37 above, wherein at least two doses are separated by a time interval. 39. The method described in 38 above, wherein the time interval is approximately 1 day to approximately 180 days. 40. The method according to 38 or 39 above, wherein the time interval is approximately 60 days to approximately 120 days. 41. The method according to any one of 38 to 40 above, wherein at least two doses are separated by a time interval of approximately 90 days. 42. The method according to any one of the above 36 to 41, comprising an RNAi agent in a dose of approximately 50 to approximately 800 mg. 43. The method according to 42 above, comprising an RNAi agent in a dose of approximately 100 to approximately 500 mg. 44. The method described in 42 or 43 above, comprising an RNAi agent in a dose of approximately 300 mg. 45. The method according to any one of 35 to 44 above, wherein the maintenance phase includes administering an RNAi agent as one or more doses. 46. ​​The method according to 45, wherein the loading phase includes administering an RNAi agent as at least two doses. 47. The method according to 46 above, wherein at least two doses are separated by a time interval. 48. The method according to 47 above, wherein at least two doses are separated by a certain time interval. 49. The method described in 48 above, wherein the time interval is between approximately one month and approximately twelve months. 50. The method described in 48 or 49 above, wherein the time interval is between approximately 3 months and approximately 9 months. 51. The method described in any of the above 48 to 50, wherein the time interval is approximately 6 months. 52. The method according to any one of 45 to 51 above, comprising an RNAi agent in a dose of approximately 50 to approximately 800 mg. 53. The method according to 52 above, comprising an RNAi agent in a dose of approximately 100 to approximately 500 mg. 54. The method according to 52 or 53 above, wherein the dose contains approximately 300 mg of RNAi agent. 55. A method for preventing the onset of atherosclerotic cardiovascular disease in a subject, comprising the step of administering a therapeutically effective dose of an interfering ribonucleic acid (RNAi) agent to the subject, RNAi is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 56. A method for treating subjects having ASCVD, a condition at risk equivalent to ASCVD, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, a need to lower LDL-C, or a combination thereof, comprising the step of administering a therapeutically effective dose of an interfering ribonucleic acid (RNAi) agent to the subject. RNAi is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 57. The method according to 55 or 56 above, wherein the subject has a baseline LDL-C level of approximately 70 mg / dl or higher. 58. The method described in any of the above 55 to 57, wherein the subject has active liver disease. 59. The method according to 58 above, wherein active liver disease is identified by one or more of the following characteristics: alanine aminotransferase (ALT) greater than twice the upper limit of normal (ULN); aspartate aminotransferase (AST) greater than twice the ULN; and total bilirubin greater than 1.5 times the ULN. 60. The method described in any of the above 55 to 57, wherein the subject does not have active liver disease. 61. The method according to 60 above, wherein active liver disease is identified by one or more of the following characteristics: alanine aminotransferase (ALT) greater than twice the upper limit of normal (ULN); aspartate aminotransferase (AST) greater than twice the ULN; and total bilirubin greater than 1.5 times the ULN. 62. The method according to any one of items 55 to 61 above, wherein the subject is being treated with background lipid-lowering therapy. 63. The method according to 62 above, wherein background lipid-lowering therapy is selected from statins, ezetimibe, and LDL apheresis. 64. The method according to 62 or 63 above, wherein background lipid-lowering therapy is maintained while the subject is receiving an RNAi agent. 65. The method described in any of items 55 to 61 above, wherein the subject has not received background lipid-lowering therapy. 66. The method described in any of 55 to 65 above, wherein the subject has a baseline triglyceride level of approximately 400 mg / dl or higher. 67. The method according to any one of 55 to 66 above, wherein the subject has a baseline estimated glomerular filtration rate (eGFR) of less than approximately 30 ml / min. 68. The method described in any of the above 55 to 67, wherein the subject has poorly controlled type 2 diabetes. 69. The method according to 68 above, wherein poorly controlled type 2 diabetes is identified by baseline glycated hemoglobin A1c levels of at least about 10%. 70. The method described in any of the above 55 to 69, wherein the subject has New York Heart Association (NYHA) class II, III, or IV heart failure. 71. The method described in any of items 55 to 70 above, wherein the ventricular ejection fraction of the subject is less than 30%. 72. The method described in any of items 55 to 71 above, wherein the subject has experienced a major adverse cardiac event within 6 months of administration of the RNAi agent. 73. The subject has experienced at least one hemorrhagic stroke, as described in any of the methods described in items 55 to 72 above. 74. The method according to any one of the above 55 to 73, wherein the subject has cardiac arrhythmia within 3 months of administration of the RNAi agent. 75. The method according to any one of items 55 to 74 above, wherein the subject does not have cardiac arrhythmias within three months of administration of RNAi agents that are not controlled by medication or resection. 76. The method described in any of items 55 to 75 above, wherein the subject does not have cardiac arrhythmia. 77. Any method described in 55 to 76 above, wherein the subject is an adult. 78. The method described in any of the above 55 to 77, wherein the subject has heterozygous familial hypercholesterolemia or homozygous familial hypercholesterolemia. 79. Any method described in any of the above 55 to 78, wherein the subject requires a reduction in LDL-C. 80. The method according to any one of the above 55 to 79, wherein low-density lipoprotein levels are reduced by more than approximately 20% compared to baseline levels and maintained for more than 15 days after administration of the RNAi agent. 81. The method according to any one of paragraphs 55 to 80 above, wherein administration of an RNAi agent reduces the level of PCSK9 by more than approximately 25% compared to the baseline level of PCSK9, and the baseline level of PCSK9 is measured before administration of the RNAi agent. 82. The method according to 81 above, wherein a reduction in PCSK9 levels of more than approximately 25% compared to baseline levels is maintained for more than 30 days after administration of the RNAi agent. 83. The method according to any one of items 55 to 82 above, wherein the administration of an RNAi agent includes a loading phase and a maintenance phase. 84. The method according to 83 above, wherein the loading phase includes administering an RNAi agent as one or more doses. 85. The method according to 84 above, wherein the loading phase includes administering an RNAi agent in at least two doses. 86. The method according to 85 above, wherein at least two doses are separated by a time interval. 87. The method described in 86 above, wherein the time interval is approximately 1 day to approximately 180 days. 88. The method described in 86 or 87 above, wherein the time interval is approximately 60 days to approximately 120 days. 89. The method described in any of the above 86 to 88, wherein the time interval is approximately 90 days. 90. The method according to any one of the above 83 to 89, comprising an RNAi agent in a dose of approximately 50 to approximately 800 mg. 91. The method according to 90 above, comprising an RNAi agent in a dose of approximately 100 to approximately 500 mg. 92. The method according to 90 or 91 above, comprising an RNAi agent in a dose of approximately 300 mg. 93. The method according to any one of 83 to 92 above, wherein the maintenance phase includes administering an RNAi agent as one or more doses. 94. The method according to 93 above, wherein the loading phase includes administering an RNAi agent in at least two doses. 95. The method according to 94 above, wherein at least two doses are separated by a time interval. 96. The method according to 95 above, wherein at least two doses are separated by a certain time interval. 97. The method according to 96 above, wherein the time interval is between approximately one month and approximately two months. 98. The method described in 96 or 97 above, wherein the time interval is between approximately 3 months and approximately 9 months. 99. The method described in any of the above 96 to 98, wherein the time interval is approximately 6 months. 100. The method according to any one of the above 93 to 99, comprising an RNAi agent in a dose of approximately 50 to approximately 800 mg. 101. The method according to 100 above, comprising an RNAi agent in a dose of approximately 100 to approximately 500 mg. 102. The method according to 100 or 101 above, comprising an RNAi agent in a dose of approximately 300 mg. 103. A method for preventing cardiac events in subjects receiving a diet designed to improve lipid levels and maximally tolerable statin therapy, wherein the subjects have heterozygous familial hypercholesterolemia and require further reduction of low-density lipoprotein cholesterol. The method includes the step of administering a prophylactically effective dose of RNAi agent. The RNAi agent is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 104. A method for preventing cardiac events in subjects receiving a diet designed to improve lipid levels and maximally tolerable statin therapy, wherein the subjects have atherosclerotic cardiovascular disease and require further reduction of low-density lipoprotein cholesterol. The method includes the step of administering a prophylactically effective dose of RNAi agent. The RNAi agent is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 105. A method for preventing cardiac events in subjects receiving dietary therapy and low-density lipoprotein-lowering therapy designed to improve lipid levels, wherein the subjects have homozygous familial hypercholesterolemia and require further reduction of low-density lipoprotein cholesterol. The method includes the step of administering a prophylactically effective dose of RNAi agent. The RNAi agent is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 106. A method for treating a subject receiving dietary therapy and maximally tolerable statin therapy designed to improve lipid levels, wherein the subject has heterozygous familial hypercholesterolemia and requires further reduction of low-density lipoprotein cholesterol. The method includes the step of administering a therapeutically effective amount of RNAi agent. The RNAi agent is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 107. A method for treating a subject receiving a diet designed to improve lipid levels and maximally tolerable statin therapy, wherein the subject has atherosclerotic cardiovascular disease and requires further reduction of low-density lipoprotein cholesterol. The method includes the step of administering a therapeutically effective amount of RNAi agent. The RNAi agent is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. 108. A method for treating a subject receiving dietary therapy and low-density lipoprotein-lowering therapy designed to improve lipid levels, wherein the subject has homozygous familial hypercholesterolemia and requires further reduction of low-density lipoprotein cholesterol. The method includes the step of administering a therapeutically effective amount of RNAi agent. The RNAi agent is a double-stranded ribonucleic acid comprising a sense strand and an antisense strand that form a double-stranded region, wherein 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. [Sequence Listing Free Text]

[0196] Sequence ID 1 5'-ACAAAAGCAAAACAGGUCUAGAA-3' Sequence ID 2 5'-CUAGACCUGUTUUGCUUUUGU-3' Sequence ID 3 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3', where a, g, c and u are 2'-O-methyl Lu(2'-OMe)A, G, C, or U; Af, Gf, Cf, or Uf are 2'-FluoroA, G, C, or U; dT is 2'-deoxythymidine; s is a phosphorothioate linkage. Sequence ID 4 5'-csusagacCfuGfudTuugcuuuugu-3', where a, g, c, and u are 2'-O-methyl(2'-OMe)A , G, C, or U; Af, Gf, Cf, or Uf are 2'-fluoroA, G, C, or U; dT is 2'- It is oxythymidine; s is phosphorothioate linked.

Claims

1. An injection device for subcutaneous administration containing a fixed dose of interferential ribonucleic acid (RNAi) agent of 275 mg to 325 mg for the prevention of cardiovascular events in human subjects, The RNAi agent comprises a double-stranded ribonucleic acid including a sense strand and an antisense strand forming a double-stranded region, the antisense strand comprising the nucleotide sequence 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO: 3), and the sense strand comprising the nucleotide sequence 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO: 4), where a, g, c, and u are 2'-O-methyl(2'-OMe)A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoroA, G, C, and U, respectively; dT is 2'-deoxythymidine; s is a phosphorothioate linkage, and the double-stranded ribonucleic acid has a tribranched GalNAc ligand covalently bonded to the 3' end of the sense strand. The subcutaneous administration includes a loading phase and a maintenance phase, wherein the maintenance phase includes an interval between administrations that is longer than the interval between loading phases. The maintenance phase comprises at least two doses, where the longer interval is a fixed time interval of 6 months between each dose. The subjects are those with atherosclerotic cardiovascular disease (ASCVD), a condition with a risk equivalent to ASCVD, an increased risk of cardiovascular disease, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, or those who need to lower low-density lipoprotein cholesterol (LDL-C), or a combination thereof. Injection device.

2. An injection device for subcutaneous administration containing a fixed dose of 275 mg to 325 mg of interfering ribonucleic acid (RNAi) agent, for the purpose of reducing cardiovascular mortality and morbidity in human subjects, The RNAi agent comprises a double-stranded ribonucleic acid including a sense strand and an antisense strand forming a double-stranded region, the antisense strand comprising the nucleotide sequence 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO: 3), and the sense strand comprising the nucleotide sequence 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO: 4), where a, g, c, and u are 2'-O-methyl(2'-OMe)A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoroA, G, C, and U, respectively; dT is 2'-deoxythymidine; s is a phosphorothioate linkage, and the double-stranded ribonucleic acid has a tribranched GalNAc ligand covalently bonded to the 3' end of the sense strand. The subcutaneous administration includes a loading phase and a maintenance phase, wherein the maintenance phase includes an interval between administrations that is longer than the interval between loading phases. The maintenance phase comprises at least two doses, where the longer interval is a fixed time interval of 6 months between each dose. The subjects are those with atherosclerotic cardiovascular disease (ASCVD), a condition with a risk equivalent to ASCVD, an increased risk of cardiovascular disease, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, or those who need to lower low-density lipoprotein cholesterol (LDL-C), or a combination thereof. Injection device.

3. An injection device for subcutaneous administration containing a fixed dose of interferential ribonucleic acid (RNAi) agent in the range of 275 mg to 325 mg, for the purpose of preventing the development of atherosclerotic cardiovascular disease in human subjects, The RNAi agent comprises a double-stranded ribonucleic acid including a sense strand and an antisense strand forming a double-stranded region, the antisense strand comprising the nucleotide sequence 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO: 3), and the sense strand comprising the nucleotide sequence 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO: 4), where a, g, c, and u are 2'-O-methyl(2'-OMe)A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoroA, G, C, and U, respectively; dT is 2'-deoxythymidine; s is a phosphorothioate linkage, and the double-stranded ribonucleic acid has a tribranched GalNAc ligand covalently bonded to the 3' end of the sense strand. The subcutaneous administration includes a loading phase and a maintenance phase, wherein the maintenance phase includes an interval between administrations that is longer than the interval between loading phases. The maintenance phase comprises at least two doses, where the longer interval is a fixed time interval of 6 months between each dose. Injection device.

4. The injection device according to any one of claims 1 to 3, wherein the fixed dose comprises 300 mg of an RNAi agent.

5. An injection device according to any one of claims 1 to 4, wherein the RNAi agent is shown in the following schematic diagram.

6. The injection device according to any one of claims 3 to 5, wherein the subject has atherosclerotic cardiovascular disease (ASCVD), a condition with a risk equivalent to ASCVD, an increased risk of cardiovascular disease, heterozygous familial hypercholesterolemia, or homozygous familial hypercholesterolemia, or needs to lower low-density lipoprotein cholesterol (LDL-C), or has a combination thereof.

7. An injection device according to any one of claims 1 to 6, wherein the subject has a baseline low-density lipoprotein cholesterol (LDL-C) level of 70 mg / dl or higher.

8. An injection device according to any one of claims 1 to 7, wherein the subject does not have active liver disease.

9. An injection device according to any one of claims 1 to 7, wherein the subject has active liver disease.

10. The injection device according to claim 8 or 9, wherein active liver disease is identified by one or more of the following characteristics: alanine aminotransferase (ALT) greater than twice the upper limit of normal (ULN); aspartate aminotransferase (AST) greater than twice the ULN; and total bilirubin greater than 1.5 times the ULN.

11. The injection device according to any one of claims 1 to 10, wherein the subject is being treated with background lipid-lowering therapy, and the background lipid-lowering therapy is selected from statins, ezetimibe, and LDL apheresis.

12. The injection device according to claim 11, wherein the background lipid-lowering therapy is a statin.

13. The injection device according to claim 11 or 12, wherein the subject is receiving the maximum acceptable statin therapy.

14. The injection device according to any one of claims 1 to 13, wherein the target has one or more of the following features: a) The subjects have a baseline triglyceride level of 400 mg / dl or less. b) The subject has a baseline estimated glomerular filtration rate (eGFR) of at least 30 ml / min, c) The subjects do not have type 2 diabetes that is not well managed. d) The subject does not have New York Heart Association (NYHA) class II, III, or IV heart failure. e) The most recent known ventricular ejection fraction of the subject is 30% or greater. f) The subjects have not experienced a major adverse cardiac event within 6 months of receiving the RNAi agent. g) The subject has not experienced a hemorrhagic stroke. h) The subject does not have cardiac arrhythmias within 3 months of receiving RNAi agents. i) The subject does not have cardiac arrhythmias within 3 months of administration of RNAi agents that are not controlled by medication or surgery. j) The subject does not have cardiac arrhythmias. k) The subject is an adult. l) The subject has heterozygous familial hypercholesterolemia or homozygous familial hypercholesterolemia, m) The subject requires a reduction in LDL-C.

15. Poorly controlled type 2 diabetes is associated with at least 10% baseline glycated hemoglobin A1c An injection device according to claim 14, specified by level.

16. An injection device according to any one of claims 1 to 15, wherein the RNAi agent reduces the level of LDL-C in the target.

17. An injection device according to any one of claims 1 to 16, wherein the reduction in LDL-C levels is greater than 20% compared to baseline levels and is maintained for more than 15 days after administration of the RNAi agent.

18. An injection device according to any one of claims 1 to 17, wherein the RNAi agent reduces the level of PCSK9 in the target.

19. The injection device according to any one of claims 1 to 18, wherein the reduction in PCSK9 levels is greater than 25% compared to baseline levels and is maintained for more than 30 days after administration of the RNAi agent, and the baseline level of PCSK9 is measured before administration of the RNAi agent.

20. The injection device according to any one of claims 1 and 4 to 19, wherein the cardiovascular event is selected from the group consisting of cardiovascular death, non-fatal myocardial infarction, severe recurrent ischemia, stroke, and symptomatic pulmonary embolism.

21. The injection device according to any one of claims 1 to 20, wherein the subject has one or more risk factors selected from the group consisting of having acute coronary syndrome; having a history of myocardial infarction, stable or unstable angina, revascularization of the coronary or other arteries, stroke, transient ischemic attack, or peripheral artery disease of atherosclerosis origin; being male; having a family history of heart disease, ASCVD, or a condition with a risk equivalent to ASCVD; being a smoker; being physically inactive; having hypertension; having high blood cholesterol; having diabetes or a prediabetic condition; being overweight or obese; having a history of pre-eclampsia during pregnancy; being unable to manage stress and / or anger; being postmenopausal; having an unhealthy diet; being 55 years of age or older; having sleep apnea; and being anemic.

22. The injectable device according to any one of claims 1 to 3, wherein the subject is receiving a dietary therapy designed to improve lipid levels and maximally tolerable statin therapy, and the subject has heterozygous familial hypercholesterolemia and requires further reduction of low-density lipoprotein cholesterol.

23. The injectable device according to any one of claims 1 to 3, wherein the subject is receiving a dietary therapy designed to improve lipid levels and maximally tolerable statin therapy, and the subject has atherosclerotic cardiovascular disease and requires further reduction of low-density lipoprotein cholesterol.

24. The injection device according to any one of claims 1 to 3, wherein the subject is receiving dietary therapy and low-density lipoprotein-lowering therapy designed to improve lipid levels, and the subject has homozygous familial hypercholesterolemia and requires further reduction of low-density lipoprotein cholesterol.

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