DIPEPTIDYL PEPTIDASE 4 (DPP4) IRNA COMPOSITIONS AND METHODS OF USE THEREOF
The present invention relates to RNAi agents, e.g., dsRNA agents, targeting the dipeptidyl peptidase 4 (DPP4) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of a DPP4 gene and to methods of treating or preventing a DPP4-associated disease, such as metabolic diseases, e.g., diabetes or lipid metabolism diseases, in a subject.
1 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of Dipeptidyl peptidase 4 (DPP4) in a cell,
(a) wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of a portion of the nucleotide sequence of any one of SEQ ID NOs:1-15, or a nucleotide sequence having at least 90% nucleotide sequence identity to a portion of the nucleotide sequence of any one of SEQ ID NOs:1-15, and the antisense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of the corresponding portion of the nucleotide sequence of any one of SEQ ID NOs:16-30, or a nucleotide sequence having at least 90% nucleotide sequence identity to a portion of the nucleotide sequence of an one of SEQ ID NOs:16-30; and
wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties;
(b) wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises a region complementary to part of an mRNA encoding a DPP4 gene (any one of SEQ ID NOs:1-15), wherein each strand independently is 14 to 30 nucleotides in length; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties; or
(c) wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense nucleotide sequences in any one of Tables 2-3 and 5-6, wherein each strand independently is 14 to 30 nucleotides in length; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties; or
(d) wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequence of nucleotides 1204-1226, 1208-1230, 1209-1231, 1210-1232, 1211-1233, 1212-1234, 1700-1722, 2223-2245, 2224-2246, 2225-2247, or 3232-3254 of SEQ ID NO:6, and the antisense strand comprises at least 15 contiguous nucleotides from the corresponding nucleotide sequence of SEQ ID NO:21, and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties; or
(e) wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense nucleotide sequences in any one of Tables 2-3 and 5-6, wherein each strand independently is 14 to 30 nucleotides in length.
2 . (canceled)
3 . (canceled)
4 . The dsRNA agent of claim 1 ,
(a) wherein the sense strand or the antisense strand is a sense strand or an antisense strand selected from the group consisting of any of the sense strands and antisense strands in any one of Tables 2-3 and 5-6; and/or
(b) wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from any one of the antisense strand nucleotide sequences of a duplex selected from the group consisting of AD-1286365.1, AD-1286369.1, AD-1286370.1, AD-1286371.1, AD-1286372.1, AD-1286373.1, AD-1286829.1, AD-1287272.1, AD-1287273.1, AD-1287274.1, and AD-1288171.1; and/or
(c) wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of nucleotides 2224-2246 of SEQ ID NO:6, and the antisense strand comprises at least 15 contiguous nucleotides from the corresponding nucleotide sequence of SEQ ID NO:21; and/or
(d) wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from the antisense strand nucleotide sequences of duplex AD-1287273.1; and/or
(e) wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of nucleotides 1211-1233 of SEQ ID NO:6, and the antisense strand comprises at least 15 contiguous nucleotides from the corresponding nucleotide sequence of SEQ ID NO:21; and/or
(f) wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from the antisense strand nucleotide sequences of duplex AD-1286372.1.
5 .- 16 . (canceled)
17 . The dsRNA agent of claim 1 , wherein the dsRNA agent comprises at least one modified nucleotide.
18 . (canceled)
19 . (canceled)
20 . The dsRNA agent of claim 17 , wherein at least one of the modified nucleotides is selected from the group a deoxy-nucleotide, a 3′-terminal deoxythimidine (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, 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 phosphonate, 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, a 2′-0 hexadecyl nucleotide, a nucleotide comprising a 2′-phosphate, a cytidine-2′-phosphate nucleotide, a guanosine-2′-phosphate nucleotide, a 2′-O-hexadecyl-cytidine-3′-phosphate nucleotide, a 2′-O-hexadecyl-adenosine-3′-phosphate nucleotide, a 2′-O-hexadecyl-guanosine-3′-phosphate nucleotide, a 2′-O-hexadecyl-uridine-3′-phosphate nucleotide, a a 5′-vinyl phosphonate (VP), a 2′-deoxyadenosine-3′-phosphate nucleotide, a 2′-deoxycytidine-3′-phosphate nucleotide, a 2′-deoxyguanosine-3′-phosphate nucleotide, a 2′-deoxythymidine-3′-phosphate nucleotide, a 2′-deoxyuridine nucleotide, and a terminal nucleotide linked to a cholesteryl derivative and a dodecanoic acid bisdecylamide group; and combinations thereof.
21 .- 23 . (canceled)
24 . The dsRNA agent of claim 20 , further comprising at least one phosphorothioate internucleotide linkage.
25 . (canceled)
26 . The dsRNA agent of claim 1 , wherein each strand is no more than 30 nucleotides in length.
27 .- 37 . (canceled)
38 . The dsRNA agent of claim 1 , wherein one or more lipophilic moieties are conjugated to one or more internal positions on at least one strand.
39 .- 47 . (canceled)
48 . The dsRNA agent of claim 1 , wherein the one or more lipophilic moieties are conjugated to one or more of the internal positions selected from the group consisting of positions 4-8 and 13-18 on the sense strand, and positions 6-10 and 15-18 on the antisense strand, counting from the 5′-end of each strand.
49 .- 57 . (canceled)
58 . The dsRNA agent of claim 1 , wherein the lipophilic moiety contains a saturated or unsaturated C4-C30 hydrocarbon chain, and an optional functional group selected from the group consisting of hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide, and alkyne.
59 .- 67 . (canceled)
68 . The dsRNA agent of claim 1 , further comprising a targeting ligand that targets a liver tissue.
69 .- 74 . (canceled)
75 . The dsRNA agent of claim 1 , further comprising a phosphate or phosphate mimic at the 5′-end of the antisense strand.
76 .- 78 . (canceled)
79 . An isolated cell containing the dsRNA agent of claim 1 .
80 . A pharmaceutical composition for inhibiting expression of a DPP4 gene, comprising the dsRNA agent of claim 1 .
81 . (canceled)
82 . A device for oral inhalative administration comprising the dsRNA agent of claim 1 .
83 . (canceled)
84 . An in vitro method of inhibiting expression of a DPP4 gene in a cell, the method comprising:
(a) contacting the cell with the dsRNA agent of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the DPP4 gene, thereby inhibiting expression of the DPP4 gene in the cell.
85 .- 87 . (canceled)
88 . A method of treating a subject having a dipeptidyl peptidase 4-(DPP4-) associated disease or a subject at risk of developing a DPP4-associated disease, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of claim 1 .
89 . The method of claim 88 , wherein the subject is a human.
90 . The method of claim 88 , wherein the DPP4-associated disease is a metabolic disease.
91 .- 96 . (canceled)
97 . The method of claim 88 , wherein the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 50 mg/kg.
98 . The method of claim 88 , wherein the dsRNA agent is administered to the subject subcutaneously: intravenously: orally: or by pulmonary system administration.
99 .- 101 . (canceled)
102 . The method of claim 88 , further comprising administering to the subject an additional agent or a therapy suitable for treatment or prevention of a DPP4-associated disorder.
103 . (canceled)
104 . (canceled)