Granted Patent B2

US 12,709,751 · App. 17/800,471 · Granted Aug 18, 2026

Inventors: James D. Mcininch (Burlington, MA); Anna Borodovsky (Melrose, MA)

Assignee: Alnylam Pharmaceuticals, Inc.

C12N15/1137C12Y304/21104C12N2310/14C12N2310/315C12N2310/321C12N2310/322C12N2310/351C12N2320/30

Abstract

Claims (83)

1 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of mannan binding lectin serine peptidase 2 (MASP2) in a cell, wherein said dsRNA comprises a sense strand and an antisense strand forming a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding MASP2, and wherein the antisense strand comprises the nucleotide sequence set forth as SEQ ID NO: 93, 92, 100, 94, 96, 70, 71, 72, 76, or 83, and the sense strand comprises the nucleotide sequence set forth as SEQ ID NO: 47, 46, 54, 48, 50, 24, 25, 26, 30, or 37 that corresponds to the antisense sequence.

2 . The dsRNA agent of claim 1 , wherein

(a) the dsRNA agent comprises at least one modified nucleotide;

(b) substantially all of the nucleotides of the sense strand comprise a modification, substantially all of the nucleotides of the antisense strand comprise a modification, or substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand comprise a modification;

(c) all of the nucleotides of the sense strand comprise a modification, all of the nucleotides of the antisense strand comprise a modification, or all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification;

(d) the dsRNA agent comprises at least one modified nucleotide selected from the group consisting of a deoxy-nucleotide, a 3′-terminal deoxythymidine (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′-hydroxyl-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a nucleotide comprising a phosphoramidate, a nucleotide comprising a non-natural nucleotide base, a tetrahydropyran modified nucleotide, a 1,5-anhydrohexitol modified nucleotide, a cyclohexenyl modified nucleotide, a nucleotide comprising a phosphorothioate group, a nucleotide comprising a methylphosphonate group, a nucleotide comprising a 5′-phosphate, a nucleotide comprising a 5′-phosphate mimic, a thermally destabilizing nucleotide, a glycol modified nucleotide (GNA), and a 2-O—(N-methylacetamide) modified nucleotide, and combinations thereof;

(e) the dsRNA agent comprises at least one modified nucleotide selected from the group consisting of LNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-alkyl, 2′-O-allyl, 2′-C-allyl, 2′-fluoro, 2′-deoxy, 2′-hydroxyl, and glycol, and combinations thereof;

(f) the dsRNA agent comprises at least one modified nucleotide selected from the group consisting of a deoxy-nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a GNA, and, a vinyl-phosphonate nucleotide, and combinations thereof;

(g) the dsRNA agent comprises at least one modified nucleotide, wherein the modified nucleotide is a thermally destabilizing nucleotide; and/or

(h) the dsRNA agent comprises at least one modified nucleotide, wherein the modified nucleotide is a thermally destabilizing nucleotide modification, and wherein the thermally destabilizing nucleotide modification is selected from the group consisting of an abasic modification, a mismatch with the opposing nucleotide in the duplex, a destabilizing sugar modification, a 2′-deoxy modification, an acyclic modification, an unlocked nucleic acid (UNA), and GNA.

3 . The dsRNA agent of claim 1 , wherein

(a) the double stranded region is 23-27 nucleotide pairs in length;

(b) wherein the double stranded region is 21-23 nucleotide pairs in length;

(c) each strand is independently no more than 30 nucleotides in length;

(d) the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length;

(e) the region of complementarity is at least 17 nucleotides in length;

(f) the region of complementarity is between 19 and 23 nucleotides in length;

(g) the region of complementarity is 19 nucleotides in length;

(h) at least one strand comprises a 3′ overhang of at least 1 nucleotide; and/or

(i) at least one strand comprises a 3′ overhang of at least 2 nucleotides.

4 . The dsRNA agent of claim 1 , further comprising a ligand.

5 . The dsRNA agent of claim 4 , wherein

(a) the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent;

(b) the ligand is an N-acetylgalactosamine (GalNAc) derivative;

(c) the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent branched linker; and/or

(d) the ligand is

6 . The dsRNA agent of claim 1 , wherein the dsRNA agent is conjugated to a ligand as shown in the following schematic

and, wherein X is O or S.

7 . The dsRNA agent of claim 1 , wherein

(a) the dsRNA agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage at the 3′-terminus and/or 5′-terminus of the sense strand and/or the antisense strand; and/or

(b) the base pair at the 1 position of the 5′-end of the antisense strand of the duplex is an AU base pair.

8 . A cell containing the dsRNA agent of claim 1 .

9 . A pharmaceutical composition for inhibiting expression of a gene encoding MASP2 comprising the dsRNA agent of claim 2 .

10 . The pharmaceutical composition of claim 9 , wherein

(a) the dsRNA agent is in an unbuffered solution;

(b) the dsRNA agent is in an unbuffered solution, wherein the unbuffered solution is saline or water;

(c) the dsRNA agent is in a buffer solution;

(d) the dsRNA agent is in a buffer solution, wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof; or

(e) the dsRNA agent is in a buffer solution, wherein the buffer solution is phosphate buffered saline (PBS).

11 . A method of inhibiting expression of a MASP2 gene in a cell, the method comprising introducing into the cell the dsRNA agent of claim 1 , thereby inhibiting expression of the MASP2 gene in the cell.

12 . The method of claim 11 , wherein

(a) the cell is within a subject;

(b) the cell is within a subject, wherein the subject is a human;

(c) the cell is within a subject, wherein the subject has a MASP2-associated disorder;

(d) the cell is within a subject having a MASP2-associated disorder, wherein the MASP2-associated disorder is selected from the group consisting of arthritis, IgA nephropathy, thrombotic microangiopathy, diabetic nephropathy and membranous nephropathy;

(e) introducing the dsRNA agent into the cell inhibits the expression of MASP2 by at least 50%, 60%, 70%, 80%, 90%, or 95%; and/or

(f) inhibiting expression of MASP2 decreases MASP2 protein level in serum of the subject by at least 50%, 60%, 70%, 80%, 90%, or 95%.

13 . A method of treating a subject having a disorder that would benefit from reduction in MASP2 expression, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of claim 1 , thereby treating the subject having the disorder that would benefit from reduction in MASP2 expression.

14 . A method of preventing at least one symptom in a subject having a disorder that would benefit from reduction in MASP2, comprising administering to the subject a prophylactically effective amount of the dsRNA agent of claim 1 , thereby preventing at least one symptom in the subject having the disorder that would benefit from reduction in MASP2 expression.

15 . The method of claim 13 , wherein

(a) the disorder is a MASP2-associated disorder;

(b) the disorder is a MASP2-associated disorder selected from the group consisting of arthritis, IgA nephropathy, thrombotic microangiopathy, diabetic nephropathy and membranous nephropathy;

(c) the subject is human;

(d) the administration of the agent to the subject causes a decrease in inflammation and/or a decrease in proteinuria;

(e) the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 50 mg/kg;

(f) the dsRNA agent is administered to the subject subcutaneously;

(g) the method further comprises determining the level of MASP2 in a sample(s) from the subject;

(h) the method further comprises determining the MASP2 protein level in a blood or serum sample(s); and/or

(i) the method further comprises administering to the subject an additional therapeutic agent for treatment of inflammation.

16 . A kit comprising the dsRNA agent of claim 1 .

17 . A vial comprising the dsRNA agent of claim 1 .

18 . A syringe comprising the dsRNA agent of claim 1 .

19 . The method of claim 14 , wherein

(a) the disorder is a MASP2-associated disorder;

(b) the disorder is a MASP2-associated disorder selected from the group consisting of arthritis, IgA nephropathy, thrombotic microangiopathy, diabetic nephropathy and membranous nephropathy;

(c) the subject is human;

(d) the administration of the agent to the subject causes a decrease in inflammation and/or a decrease in proteinuria;

(e) the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 50 mg/kg;

(f) the dsRNA agent is administered to the subject subcutaneously;

(g) the method further comprises determining the level of MASP2 in a sample(s) from the subject;

(h) the method further comprises determining the level of MASP2 protein level in a blood or serum sample(s); and/or

(i) the method further comprises administering to the subject an additional therapeutic agent for treatment of inflammation.

20 . The dsRNA of claim 1 , wherein

(a) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 139 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 185;

(b) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 138 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 184;

(c) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 146 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 192;

(d) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 140 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 186;

(e) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 142 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 188;

(f) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 116 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 162;

(g) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 117 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 163;

(h) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 118 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 164;

(i) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 122 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 168; or

(j) the sense strand comprises the sequence and all of the modifications of SEQ ID NO: 129 and the antisense strand comprises the sequence and all of the modifications of SEQ ID NO: 175.

Assignments (2)

Continuity (2)

References Cited (400)