Methods and compositions for treating a proprotein convertase subtilisin kexin (pcsk9) gene-associated disorder

Abstract

The invention relates to methods of inhibiting the expression of a PCSK9 gene in a subject, as well as therapeutic and prophylactic methods for treating subjects having a lipid disorder, such as a hyperlipidemia using RNAi agents, e.g., double-stranded RNAi agents, targeting the PCSK9 gene.

1 . A method of inhibiting the expression of a Proprotein convertase subtilisin kexin 9 (PCSK9) gene in a human subject, comprising subcutaneously administering to the subject a double stranded ribonucleic acid (RNAi) agent, or salt thereof, in a dosing regimen that includes a loading phase followed by a maintenance phase,
 wherein the loading phase comprises administering a fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject and wherein the maintenance phase comprises administering the same fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject,   wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises the nucleotide sequence of 5′-csusagacCfuGfudTuugcuuuugu-3′ (SEQ ID NO: 687) and the antisense strand comprises the nucleotide sequence of 5′-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3′ (SEQ ID NO: 688),   wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, and U, respectively; dT is 2′-deoxythymidine; and s is a phosphorothioate linkage, and   wherein the double stranded RNAi agent is conjugated to a N-acetylgalactosamine (GalNAc)3 ligand.   
     
     
         2 . A method of treating a human subject having a hyperlipidemia, comprising subcutaneously administering to the subject a double stranded ribonucleic acid (RNAi) agent, or salt thereof, in a dosing regimen that includes a loading phase followed by a maintenance phase,
 wherein the loading phase comprises administering a fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject and wherein the maintenance phase comprises administering the same fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject,   wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises the nucleotide sequence of 5′-csusagacCfuGfudTuugcuuuugu-3′ (SEQ ID NO: 687) and the antisense strand comprises the nucleotide sequence of 5′-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3′ (SEQ ID NO: 688),   wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, and U, respectively; dT is 2′-deoxythymidine; and s is a phosphorothioate linkage, and   wherein the double stranded RNAi agent is conjugated to a N-acetylgalactosamine (GalNAc)3 ligand.   
     
     
         3 .- 12 . (canceled) 
     
     
         13 . The method of  claim 1 , further comprising administering an additional therapeutic agent to the subject. 
     
     
         14 . The method of  claim 2 , further comprising administering an additional therapeutic agent to the subject. 
     
     
         15 . The method of  claim 2 , wherein the hyperlipidemia is hypercholesterolemia. 
     
     
         16 . The method of  claim 1 , wherein the double stranded RNAi agent or salt thereof is administered in a pharmaceutical composition. 
     
     
         17 . The method of  claim 2 , wherein the double stranded RNAi agent or salt thereof is administered in a pharmaceutical composition. 
     
     
         18 . The method of  claim 1 , wherein the (GalNAc)3 ligand is 
       
         
           
           
               
               
           
         
       
     
     
         19 . The method of  claim 2 , wherein the (GalNAc)3 ligand is 
       
         
           
           
               
               
           
         
       
     
     
         20 . The method of  claim 18 , wherein the double stranded RNAi agent is conjugated to the (GalNAc)3 ligand as shown in the following schematic 
       
         
           
           
               
               
           
         
         wherein X is O or S. 
       
     
     
         21 . The method of  claim 19 , wherein the double stranded RNAi agent is conjugated to the (GalNAc)3 ligand as shown in the following schematic 
       
         
           
           
               
               
           
         
         wherein X is O or S. 
       
     
     
         22 . The method of  claim 1 , wherein the sense strand comprises the nucleotide sequence of 5′-csusagacCfuGfudTuugcuuuuguL96-3′ (SEQ ID NO: 687) and the antisense strand comprises the nucleotide sequence of 5′-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3′ (SEQ ID NO: 688), wherein a, g, and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, and U, respectively; dT is 2′-deoxythymidine; s is a phosphorothioate linkage; and L96 is 
       
         
           
           
               
               
           
         
         wherein L96 is conjugated to the 3′ end of the sense strand as shown in the following schematic: 
       
       
         
           
           
               
               
           
         
       
       wherein X is O. 
     
     
         23 . The method of  claim 2 , wherein the sense strand comprises the nucleotide sequence of 5′-csusagacCfuGfudTuugcuuuuguL96-3′ (SEQ ID NO: 687) and the antisense strand comprises the nucleotide sequence of 5′-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3′ (SEQ ID NO: 688), wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, and U, respectively; dT is 2′-deoxythymidine; s is a phosphorothioate linkage; and L96 is 
       
         
           
           
               
               
           
         
         wherein L96 is conjugated to the 3′ end of the sense strand as shown in the following schematic: 
       
       
         
           
           
               
               
           
         
         wherein X is O. 
       
     
     
         24 . The method of  claim 22 , wherein the double stranded RNAi agent is in a salt form. 
     
     
         25 . The method of  claim 23 , wherein the double stranded RNAi agent is in a salt form. 
     
     
         26 . A method of decreasing levels of low density lipoprotein (LDLc) in serum of a human subject having a hyperlipidemia, comprising subcutaneously administering to the subject a double stranded ribonucleic acid (RNAi) agent, or salt thereof, in a dosing regimen that includes a loading phase followed by a maintenance phase,
 wherein the loading phase comprises administering a fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject and wherein the maintenance phase comprises administering the same fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject,   wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises the nucleotide sequence of 5′-csusagacCfuGfudTuugcuuuugu-3′ (SEQ ID NO: 687) and the antisense strand comprises the nucleotide sequence of 5′-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3′ (SEQ ID NO: 688),   wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, and U, respectively; dT is 2′-deoxythymidine; and s is a phosphorothioate linkage, and   wherein the double stranded RNAi agent is conjugated to a N-acetylgalactosamine (GalNAc)3 ligand.   
     
     
         27 . The method of  claim 26 , wherein the subject has hypercholesterolemia. 
     
     
         28 . The method of  claim 26 , wherein the subject has heterozygous LDL receptor genotype. 
     
     
         29 . A method of decreasing levels of low density lipoprotein (LDLc) in serum of a human subject having risk factors associated with cardiovascular disease, comprising subcutaneously administering to the subject a double stranded ribonucleic acid (RNAi) agent, or salt thereof, in a dosing regimen that includes a loading phase followed by a maintenance phase,
 wherein the loading phase comprises administering a fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject and wherein the maintenance phase comprises administering the same fixed dose of 275 mg to 325 mg of the RNAi agent, or salt thereof, to the subject,   wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises the nucleotide sequence of 5′-csusagacCfuGfudTuugcuuuugu-3′ (SEQ ID NO: 687) and the antisense strand comprises the nucleotide sequence of 5′-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3′ (SEQ ID NO: 688),   wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, and U, respectively; dT is 2′-deoxythymidine; and s is a phosphorothioate linkage, and   wherein the double stranded RNAi agent is conjugated to a N-acetylgalactosamine (GalNAc)3 ligand,   wherein said one or more risk factors comprise diabetes, previous personal history of coronary heart disease (CHD) or noncoronary atherosclerosis, family history of cardiovascular disease, tobacco use, hypertension and/or obesity.   
     
     
         30 . The method of  claim 29 , wherein the risk factors are associated with arteriosclerosis, heart valve disease, arrhythmia, heart failure, hypertension, orthostatic hypotension, shock, endocarditis, diseases of the aorta and its branches, disorders of the peripheral vascular system, heart attack, cardiomyopathy, and congenital heart disease. 
     
     
         31 . The method of  claim 1 , wherein the fixed dose administered to the subject is about 300 mg. 
     
     
         32 . The method of  claim 2 , wherein the fixed dose administered to the subject is about 300 mg. 
     
     
         33 . The method of  claim 1 , wherein the maintenance phase comprises administering the fixed dose of the double stranded RNAi agent, or salt thereof, to the subject once every six months. 
     
     
         34 . The method of  claim 2 , wherein the maintenance phase comprises administering the fixed dose of the double stranded RNAi agent, or salt thereof, to the subject once every six months. 
     
     
         35 . The method of  claim 1 , wherein the loading phase comprises administering a fixed dose of 300 mg of the RNAi agent, or salt thereof, to the subject and wherein the maintenance phase comprises administering the same fixed dose of 300 mg of the RNAi agent, or salt thereof, to the subject once every six months. 
     
     
         36 . The method of  claim 2 , wherein the loading phase comprises administering a fixed dose of 300 mg of the RNAi agent, or salt thereof, to the subject and wherein the maintenance phase comprises administering the same fixed dose of 300 mg of the RNAi agent, or salt thereof, to the subject every six months.