AMYLOID PRECURSOR PROTEIN (APP) RNAi AGENT COMPOSITIONS AND METHODS OF USE THEREOF

Abstract

The disclosure relates to double stranded ribonucleic acid (dsRNAi) agents and compositions targeting the APP gene, as well as methods of inhibiting expression of an APP gene and methods of treating subjects having an APP-associated disease or disorder, such as cerebral amyloid angiopathy (CAA) and early onset familial Alzheimer disease (EOFAD or eFAD), using such dsRNAi agents and compositions.

1 - 168 . (canceled) 
     
     
         169 . A double stranded ribonucleic acid (RNAi) agent comprising a sense strand and an antisense strand, wherein
 (a) the antisense strand comprises a region of complementarity comprising at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence:   
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 2743) 
                 
                     
                   5′-UUAGGUTGGAUTUUCGUAGCCGU-3′ 
                 
                     
                   or 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 1870) 
                 
                     
                   5′-UUAGGUTGGAUUUUCGUAGCCGU-3′; 
                 
             
                
                
                
                
                
                
               
            
           
         
         (b) the sense strand comprises one or more lipophilic moieties conjugated to one or more non-terminal nucleotide positions, excluding the cleavage site region of the sense strand; and 
         (c) the double stranded RNAi agent comprises at least one modified nucleotide. 
       
     
     
         170 . The double stranded RNAi agent of  claim 169 , wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence 5′-GGCUACGAAAAUCCAACCUAA-3′ (SEQ ID NO: 2735). 
     
     
         171 . The double stranded RNAi agent of  claim 170 , wherein
 (a) all of the nucleotides of the sense strand are modified nucleotides;   (b) substantially all of the nucleotides of the antisense strand are modified nucleotides;   (c) all of the nucleotides of the antisense strand are modified nucleotides; or   (d) all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified nucleotides.   
     
     
         172 . The double stranded RNAi agent of  claim 170 , wherein at least one of the modified nucleotides is selected from the group consisting of a deoxy-nucleotide, a 3′-terminal deoxy-thymidine (dT) nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-O-allyl-modified nucleotide, 2′-C-alkyl-modified nucleotide, 2′-hydroxy-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a tetrahydropyran modified nucleotide, a 1,5-anhydrohexitol modified nucleotide, a cyclohexenyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a nucleotide comprising a 5′-methylphosphonate group, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic, a nucleotide comprising vinyl phosphate, a nucleotide comprising adenosine-glycol nucleic acid (GNA), a nucleotide comprising thymidine-glycol nucleic acid (GNA)S-Isomer, a nucleotide comprising 2-hydroxymethyl-tetrahydrofurane-5-phosphate, a nucleotide comprising 2′-deoxythymidine-3′phosphate, a nucleotide comprising 2′-deoxyguanosine-3′-phosphate, and a terminal nucleotide linked to a cholesteryl derivative and a dodecanoic acid bisdecylamide group. 
     
     
         173 . The double stranded RNAi agent of  claim 172 , wherein said modified nucleotide is selected from the group consisting of a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, 3′-terminal deoxy-thymidine nucleotides (dT), a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide. 
     
     
         174 . The double stranded RNAi agent of  claim 172 , wherein the modifications on the nucleotides are 2′-O-methyl, GNA, and 2′fluoro modifications. 
     
     
         175 . The double stranded RNAi agent of  claim 172 , further comprising at least one phosphorothioate internucleotide linkage. 
     
     
         176 . The double stranded RNAi agent of  claim 175 , wherein the double stranded RNAi agent comprises 6-8 phosphorothioate internucleotide linkages. 
     
     
         177 . The double stranded RNAi agent of  claim 169 , wherein the region of complementarity is at least 17 nucleotides in length. 
     
     
         178 . The double stranded RNAi agent of  claim 169 , wherein the region of complementarity is 19-23 nucleotides in length. 
     
     
         179 . The double stranded RNAi agent of  claim 169 , wherein each strand is no more than 30 nucleotides in length. 
     
     
         180 . The double stranded RNAi agent of  claim 169 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide. 
     
     
         181 . The double stranded RNAi agent of  claim 169 , wherein the antisense strand comprises a 3′ overhang of at least 2 nucleotides. 
     
     
         182 . The double-stranded RNAi agent of  claim 169 , wherein the one or more lipophilic moieties are conjugated to one or more of positions 4-8 and 13-18 on the sense strand. 
     
     
         183 . The double-stranded RNAi agent of  claim 169 , wherein one or more lipophilic moieties are conjugated to one or more of positions 5, 6, 7, 15, and 17 on the sense strand. 
     
     
         184 . The double-stranded RNAi agent of  claim 169 , wherein the lipophilic moiety contains a saturated or unsaturated C 4 -C 30  hydrocarbon chain, and an optional functional group selected from the group consisting of hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide, and alkyne. 
     
     
         185 . The double-stranded RNAi agent of  claim 184 , wherein the lipophilic moiety contains a saturated or unsaturated C6-Cis hydrocarbon chain. 
     
     
         186 . The double-stranded RNAi agent of  claim 185 , wherein the lipophilic moiety contains a saturated or unsaturated C16 hydrocarbon chain. 
     
     
         187 . The double stranded RNAi agent of  claim 169 , wherein the one or more non-terminal nucleotide positions of the sense strand have the following structure: 
       
         
           
           
               
               
           
         
       
       wherein B is a nucleotide base or a nucleotide base analog. 
     
     
         188 . The double-stranded RNAi agent  claim 169 , further comprising a phosphate or phosphate mimic at the 5′-end of the antisense strand. 
     
     
         189 . The double-stranded RNAi agent of  claim 188 , wherein the phosphate mimic is a 5′-vinyl phosphonate (VP). 
     
     
         190 . A cell containing the double stranded RNAi agent of  claim 169 . 
     
     
         191 . A pharmaceutical composition comprising the double stranded RNAi agent of  claim 169 . 
     
     
         192 . The pharmaceutical composition of  claim 191 , comprising a buffer solution. 
     
     
         193 . The pharmaceutical composition of  claim 192 , wherein said buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof. 
     
     
         194 . The pharmaceutical composition of  claim 192 , wherein the buffer solution is phosphate buffered saline (PBS). 
     
     
         195 . A method of inhibiting expression of an amyloid precursor protein (APP) gene in a cell, the method comprising:
 (a) contacting the cell with the double stranded RNAi agent of  claim 169 ; and   (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of an APP gene, thereby inhibiting expression of the APP gene in the cell.   
     
     
         196 . The method of  claim 195 , wherein said cell is within a subject. 
     
     
         197 . The method of  claim 196 , wherein the subject is a human. 
     
     
         198 . The method of  claim 197 , wherein the human subject suffers from an APP-associated disorder. 
     
     
         199 . The method of  claim 198 , wherein the APP-associated disease is cerebral amyloid angiopathy (CAA), early onset familial Alzheimer disease (EOFAD), or Alzheimer's disease (AD). 
     
     
         200 . A method of treating a human subject having an APP-associated disorder, comprising administering to the subject a therapeutically effective amount of the double stranded RNAi agent of  claim 169 , thereby treating said subject. 
     
     
         201 . The method of  claim 200 , wherein the APP-associated disease is cerebral amyloid angiopathy (CAA), early onset familial Alzheimer disease (EOFAD), or Alzheimer's disease (AD). 
     
     
         202 . The method of  claim 200 , further comprising administering an additional therapeutic agent to the subject. 
     
     
         203 . The method of  claim 200 , wherein the administering is by intrathecal administration. 
     
     
         204 . The double-stranded RNAi agent  claim 169 , selected from the group consisting of AD-454973, AD-886864.1, AD-886865.1, AD-886866.1, AD-886867.1, AD-886868.1, AD-886869.1, AD-886872.1, AD-886876.1, AD-886878.1, AD-886879.1, AD-886886.1, and AD-886887.1. 
     
     
         205 . The double-stranded RNAi agent  claim 169 , selected from the group consisting of AD-886870.1, AD-886871.1, AD-886873.1, AD-886874.1, AD-886875.1, AD-886877.1, AD-886877.2, AD-886880.1, AD-886881.1, AD-886888.1, AD-886889.1, or AD-886889.2.