DOUBLE TAPE IRNA AGENT AND USES THEREOF

Description

DOUBLE TAPE IRNA AGENT AND USES THEREOF Separated from BR112020022546-0, filed on May 7, 2019.

[001] This application claims the benefit of priority with respect to U.S. Provisional Application No. 62 / 668,072 filed on May 7, 2018; U.S. Provisional Application No. 62 / 738,747 filed on September 28, 2018; and U.S. Provisional Application No. 62 / 773,082 filed on November 29, 2018, all of which are incorporated herein by reference in their entirety. BACKGROUND

[002] Efficient delivery of an iRNA agent to cells in vivo requires specific targeting and substantial protection of the extracellular environment, particularly serum proteins. RNAi-based therapeutics show promising clinical data for the treatment of liver-associated diseases. However, the delivery of siRNA to extrahepatic tissues remains a hurdle, limiting the use of siRNA-based therapies.

[003] One of the factors limiting the experimental and therapeutic application of iRNA agents in vivo is the ability to efficiently deliver intact siRNA. Particular difficulties have been associated with non-viral gene transfer to the retina in vivo. One of the challenges is overcoming the internal limiting membrane, which prevents retinal transfection. Additionally, it has been shown that negatively charged vitreous sugars interact with positive transfection DNA-reagent complexes, promoting their aggregation, which prevents diffusion and cellular uptake.

[004] The delivery of oligonucleotides to the central nervous system (CNS) poses particular problems due to the blood-brain barrier (BBB), which free oligonucleotides cannot cross. One means of delivering oligonucleotides to the CNS is via intrathecal delivery. However, oligonucleotides also need to be efficient Petition 870260049905, dated 05 / 25 / 2026, page 15 / 728 2 / 335 effectively internalized into target cells of the CNS to achieve the desired therapeutic effect. Previous work has typically used delivery reagents such as liposomes, cationic lipids, and nanoparticles forming complexes to aid in the intracellular internalization of oligonucleotides into cells of neuronal origin.

[005] Thus there is a continuing need for new and improved methods for delivering siRNA molecules in vivo, without the use of tissue delivery reagents, to achieve and enhance the therapeutic potential of iRNA agents. SUMMARY

[006] One aspect of the invention provides a double-stranded iRNA agent comprising:

[007] an antisense strand that is complementary to a target gene; a sense strand that is complementary to said antisense strand; and one or more lipophilic portions conjugated to one or more internal positions in at least one strand, optionally via a ligand or carrier.

[008] In some embodiments, the lipophilicity of the lipophilic portion, measured by the octanol-water partition coefficient, logKow, exceeds 0. The lipophilic portion may have a logKow exceeding 1, exceeding 1.5, exceeding 2, exceeding 3, exceeding 4, exceeding 5, or exceeding 10.

[009] In some embodiments, the hydrophobicity of the double-stranded RNA agent, measured by the unbound portion in the double-stranded RNA agent plasma protein binding assay, exceeds 0.2. In one embodiment, the determined plasma protein binding assay is an electrophoretic mobility shift assay (EMSA) using human serum albumin protein. The hydrophobicity of the double-stranded RNA agent, measured by the unbound siRNA portion in the binding assay, exceeds 0.15, exceeds 0.2, exceeds 0.25, Petition 870260049905, dated 05 / 25 / 2026, p. 16 / 728 3 / 335 exceeds 0.3, exceeds 0.35, exceeds 0.4, exceeds 0.45, or exceeds 0.5 for enhanced in vivo delivery of siRNA.

[0010] In some embodiments, the lipophilic moiety is an aliphatic, cyclic compound such as alicyclic or polycyclic such as polyalicyclic, such as a steroid (e.g., sterol) or a linear or branched aliphatic hydrocarbon. Exemplary lipophilic moieties are lipid, cholesterol, retinoic acid, cholic acid, adamantanoacetic acid, 1-pyrenebutyric acid, dihydrotestosterone, 1,3-bisO(hexadecyl)glycerol, geranyloxyhexanol, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, ibuprofen, naproxen, dimethoxytrityl or phenoxazine.

[0011] Suitable lipophilic moieties also include those containing a saturated or unsaturated C4-C30 hydrocarbon chain (e.g., C4-C30 alkyl or alkenyl) θ an optional functional group selected from the group consisting of hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide, and alkyne. The functional groups are useful for attaching the lipophilic moiety to the iRNA agent. In some embodiments, the lipophilic moiety contains a saturated or unsaturated C6-C18 hydrocarbon chain (e.g., a linear C6-C18 alkyl or alkenyl). In one embodiment, the lipophilic moiety contains a saturated or unsaturated Cw hydrocarbon chain (e.g., a linear Cw alkyl or alkenyl).

[0012] In some embodiments, the lipophilic portion is a C6-C30 acid (e.g., hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, octadecanoic acid, oleic acid, linoleic acid, arachidonic acid, cis4,7,10,13,16,19-docosahexaenoic acid, vitamin A, vitamin E, cholesterol, Petition 870260049905, dated 05 / 25 / 2026, page 17 / 728 4 / 335 etc.) or a C6-C30 alcohol (e.g., hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol, tridecanol, tetradecanol, pentadecanol, hexadecanol, heptadecanol, octadecanol, oleyl alcohol, linoleyl alcohol, arachidonic alcohol, c / s-4,7,10,13,16,19-docosahexanol, retinol, vitamin E, cholesterol, etc.).

[0013] The lipophilic portion can be conjugated to the iRNA agent through direct attachment to the ribosugar of the iRNA agent. Alternatively, the lipophilic portion can be conjugated to the iRNA agent via a ligand or a carrier.

[0014] In certain embodiments, the lipophilic portion is conjugated to the iRNA agent via one or more ligands (ties).

[0015] In some embodiments, the lipophilic portion is conjugated to the double-stranded RNA agent via a linker containing an ether, thioether, urea, carbonate, amine, amide, maleimidathioether, disulfide, phosphodiester, sulfonamide linkage, a product of a click reaction (e.g., a triazole from the azide-alkyne cycloaddition) or carbamate.

[0016] In some embodiments, at least one of the linkers (amara) is a redox-cleavable linker (such as a reductively cleavable linker; for example, a disulfide group), an acid-cleavable linker (for example, a hydrazone group, an ester group, an acetal or a ketal group), an esterase-cleavable linker (for example, an ester group), a phosphatase-cleavable linker (for example, a phosphate group) or a peptidase-cleavable linker (for example, a peptide linkage).

[0017] In other embodiments, at least one of the linkers (ties) is a biocleavable linker selected from the group consisting of DNA, RNA, disulfide, amide, functionalized monosaccharides or oligosaccharides of galactosamine, glucosamine, glucose, galactose, mannose and combinations thereof. Petition 870260049905, dated 05 / 25 / 2026, page 18 / 728 5 / 335

[0018] In certain embodiments, the lipophilic moiety is conjugated to the double-stranded RNA agent via a carrier that replaces one or more nucleotide(s). The carrier may be a cyclic group or an acyclic group. In one embodiment, the cyclic group is selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl, and decalin. In one embodiment, the acyclic group is a moiety based on a serinol backbone or a diethanolamine backbone.

[0019] In some embodiments, the carrier replaces one or more nucleotide(s) in the internal position(s) of the double-stranded iRNA agent.

[0020] In other embodiments, the carrier replaces the nucleotides at the terminal end of the sense strand or antisense strand. In one embodiment, the carrier replaces the terminal nucleotide at the 3' end of the sense strand, thus functioning as a terminal cap protecting the 3' end of the sense strand. In one embodiment, the carrier is a cyclic group having an amine, for example, the carrier may be pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuranyl, or decalinyl.

[0021] In one embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, which include all positions except the two terminal positions at each end of the strand. In another embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, which include all positions except the three terminal positions at each end of the strand. Petition 870260049905, dated 05 / 25 / 2026, p. 19 / 728 6 / 335

[0022] In one embodiment, at least one lipophilic moiety is conjugated to one or more positions at at least one end of the duplex region, which includes all positions within the duplex region but does not include the protruding region or the vehicle that replaces the terminal nucleotide at the 3' end of the sense strand.

[0023] In one embodiment, at least one lipophilic moiety is conjugated to the sense strand within the first five base pairs at the 5' end of the antisense strand of the duplex region.

[0024] In one embodiment, at least one lipophilic moiety is conjugated to the sense strand within the first four base pairs at the 5' end of the antisense strand of the duplex region.

[0025] In one embodiment, at least one lipophilic moiety is conjugated to the sense strand within the first three base pairs at the 5' end of the antisense strand of the duplex region.

[0026] In one embodiment, at least one lipophilic moiety is conjugated to the sense strand within the first two base pairs at the 5' end of the antisense strand of the duplex region.

[0027] In one embodiment, at least one lipophilic moiety is conjugated to the sense strand at the first base pair at the 5' end of the antisense strand of the duplex region.

[0028] In one embodiment, the lipophilic portion is conjugated to one or more internal positions on at least one strand, which excludes the region of the cleavage site of the sense strand. For example, the internal positions exclude positions 9-12 counting from the 5' end of the sense strand. For example, the internal positions exclude positions 9-11 counting from the 5' end of the sense strand. Alternatively, the internal positions exclude positions 11-13 counting from the 3' end of the sense strand.

[0029] In one embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, which excludes the re Petition 870260049905, dated 05 / 25 / 2026, page 20 / 728 7 / 335 region of the antisense tape splitting location. For example, the inner positions exclude positions 12-14 counting from the 5' end of the antisense tape.

[0030] In one embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, excluding positions 11-13 in the sense strand, counting from the 3' end, and positions 12-14 in the antisense strand, counting from the 5' end.

[0031] In one embodiment, one or more lipophilic portions are conjugated to one or more of the following internal positions: positions 4-8 and 13-18 on the sense strip and positions 6-10 and 15-18 on the antisense strip, counting from the 5' end of each strip.

[0032] In one embodiment, one or more lipophilic portions are conjugated to one or more of the following internal positions: positions 5, 6, 7, 15 and 17 on the sense strip and positions 15 and 17 on the antisense strip, counting from the 5' end of each strip.

[0033] In some embodiments, the sense and antisense strands of the double-stranded iRNA agent are each 15 to 30 nucleotides in length.

[0034] In one embodiment, the sense and antisense strands of a double-stranded iRNA agent are each 19 to 25 nucleotides in length.

[0035] In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each 21 to 23 nucleotides in length.

[0036] In some embodiments, the double-stranded iRNA agent comprises a single-stranded overhang at at least one of the termini, for example, 3' and / or 5' overhang(s) of 1-10 nucleotides in length, for example, an overhang of 1, 2, 3, 4, 5 or 6 nucleotides. In some embodiments, both strands have at least one 1-5 (e.g., 1, 2, 3, 4 or 5) single-stranded nucleotide stretch. Petition 870260049905, dated 05 / 25 / 2026, p. 21 / 728 8 / 335 pies in the double-stranded region. In one embodiment, the single-stranded overhang is 1, 2, or 3 nucleotides in length. In some embodiments, the double-stranded iRNA agent may also have a blunt end, located at the 5' end of the antisense strand (or at the 3' end of the sense strand) or vice versa. In one embodiment, the double-stranded iRNA agent comprises a 3' overhang at the 3' end of the antisense strand and, optionally, a blunt end at the 5' end of the antisense strand. In one embodiment, the double-stranded iRNA agent has a 5' overhang at the 5' end of the sense strand and, optionally, a blunt end at the 5' end of the antisense strand. In one embodiment, the double-stranded iRNA agent has two blunt ends at both ends of the iRNA duplex.

[0037] In one embodiment, the sense strand of the double-stranded iRNA agent is 21 nucleotides long, and the antisense strand is 23 nucleotides long, wherein the strands form a double-stranded region of 21 consecutive base pairs having a single-stranded overhang 2 nucleotides long at the 3' end.

[0038] In some embodiments, the lipophilic portion is conjugated to a nucleobase, sugar moiety, or internucleoside linkage of the double-stranded iRNA agent.

[0039] In some embodiments, the double-stranded iRNA agent additionally comprises a phosphate or phosphate mimetic at the 5' end of the antisense strand. In one embodiment, the phosphate mimetic is a 5'-vinyl phosphonate (VP).

[0040] In some embodiments, the 5' end of the antisense strand of the double-stranded iRNA agent does not contain a 5'-vinyl phosphonate (VP).

[0041] In some embodiments, the double-stranded iRNA agent additionally comprises at least one chiral phosphorus atom, Petition 870260049905, dated 05 / 25 / 2026, p. 22 / 728 9 / 335 terminal.

[0042] A chiral, site-specific modification for internucleotide bonding can occur at the 5' end, 3' end, or both the 5' and 3' ends of a strand. This is referred to here as a terminal chiral modification. The terminal modification can occur at a 3' or 5' terminal position in a terminal region, for example, at a position on a terminal nucleotide or within the last 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides of a strand. A chiral modification can occur on the sense strand, antisense strand, or both the sense and antisense strands. Each of the pure chiral phosphorus atoms can be in the Rp configuration or Sp configuration and their combinations.Further details regarding chiral modifications and chirally modified dsRNA agents can be found in PCT / US18 / 67103, entitled Chirally Modified Double-Strand RNA Agents, filed December 21, 2018, which is incorporated herein by reference in its entirety.

[0043] In some embodiments, the double-stranded iRNA agent further comprises a terminal chiral modification occurring at the first internucleotide bond at the 3' end of the antisense strand, having the phosphorus atom of the bond in the sp configuration; a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the antisense strand, having the phosphorus atom of the bond in the rp configuration; and a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the ser strand, having the phosphorus atom of the bond in the rp configuration or sp configuration.

[0044] In one embodiment, the double-stranded iRNA agent further comprises a terminal chiral modification occurring at the first and second internucleotide bonds at the 3' end of the antisense strand, having the phosphorus atom of the bond in the sp configuration; Petition 870260049905, dated 05 / 25 / 2026, page 23 / 728 10 / 335 a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the antisense strand, having the phosphorus atom of the bond in the Rp configuration; and a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the strand, having the phosphorus atom of the bond in the Rp or Sp configuration.

[0045] In one embodiment, the double-stranded iRNA agent further comprises a terminal chiral modification occurring at the first, second, and third internucleotide bonds at the 3' end of the antisense strand, having the phosphorus atom of the bond in the sp configuration; a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the antisense strand, having the phosphorus atom of the bond in the rp configuration; and a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the ser strand, having the phosphorus atom of the bond in the rp or sp configuration.

[0046] In one embodiment, the double-stranded iRNA agent further comprises a terminal chiral modification occurring at the first and second internucleotide bonds at the 3' end of the antisense strand, having the phosphorus atom of the bond in the sp configuration; a terminal chiral modification occurring at the third internucleotide bond at the 3' end of the antisense strand, having the phosphorus atom of the bond in the rp configuration; a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the antisense strand, having the phosphorus atom of the bond in the rp configuration; and a terminal chiral modification occurring at the first internucleotide bond at the 5' end of the sera strand, having the phosphorus atom of the bond in the rp or sp configuration.

[0047] In one embodiment, the double-stranded iRNA agent further comprises a terminal chiral modification occurring at Petition 870260049905, dated 05 / 25 / 2026, page 24 / 728 11 / 335 first and second internucleotide bonds at the 3' end of the antisense strand, with the phosphorus atom of the bond in the Sp configuration; a terminal chiral modification occurring in the first and second internucleotide bonds at the 5' end of the antisense strand, with the phosphorus atom of the bond in the Rp configuration; and a terminal chiral modification occurring in the first internucleotide bond at the 5' end of the strand, with the phosphorus atom of the bond in the Rp or Sp configuration.

[0048] In some embodiments, the double-stranded iRNA agent has at least two phosphorothioate internucleotide bonds in the first five nucleotides in the antisense strand (counting from the 5' end).

[0049] In some embodiments, the antisense strand comprises two blocks of one, two, or three phosphorothioate internucleotide bonds separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 phosphate internucleotide bonds.

[0050] In some embodiments, the double-stranded RNA agent additionally comprises a targeting ligand that targets a receptor that mediates delivery to a specific CNS tissue. In one embodiment, the targeting ligand is selected from the group consisting of Angiopep-2, lipoprotein receptor-related protein (LRP) ligand, bEnd.3 cell-binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein, and LDL receptor ligand.

[0051] In some embodiments, the double-stranded RNA agent additionally comprises a targeting ligand that targets a receptor that mediates delivery to an ocular tissue. In one embodiment, the targeting ligand is selected from the group consisting of trans-retinol, RGD peptide, LDL receptor ligand, and carbohydrate-based ligands. In another embodiment, the di Petition 870260049905, dated 05 / 25 / 2026, page 25 / 728 12 / 335 reaction is an RGD peptide, such as H-Gly-Arg-Gly-Asp-SerPro-Lys-Cys-OH or Cyclo(-Arg-Gly-Asp-D-Phe-Cys).

[0052] In some embodiments, the double-stranded iRNA agent additionally comprises a targeting ligand that targets liver tissue. In some embodiments, the targeting ligand is a carbohydrate-based ligand. In one embodiment, the targeting ligand is a GaINAc conjugate.

[0053] All aspects and embodiments above would be applicable to an oligonucleotide having one or more lipophilic moieties conjugated to one or more internal positions in the oligonucleotide. In some embodiments, 100%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, or 30% of the oligonucleotide is modified. For example, when 50% of the oligonucleotide is modified, 50% of all nucleotides present in the oligonucleotide contain a modification as described herein.

[0054] In one embodiment, the oligonucleotide is a double-stranded dsRNA agent, and at least 50% of the nucleotides of the double-stranded dsRNA agent are independently modified with 2'-O-methyl, 2'-O-allyl, 2'-deoxy, or 2'-fluorine.

[0055] In one embodiment, the oligonucleotide is an antisense, and at least 50% of the antisense nucleotides are independently modified with LNA, CeNA, 2'-methoxyethyl, or 2'-deoxy.

[0056] In some embodiments, the double-stranded iRNA agent has fewer than 12, fewer than 10, fewer than 8, fewer than 6, fewer than 4, fewer than 2, or no 2'-F modifications on the sense strand. In some embodiments, the double-stranded iRNA agent has fewer than 12, fewer than 10, fewer than 8, fewer than 6, fewer than 4, fewer than 2, or no 2'-F modifications on the antisense strand.

[0057] In some embodiments, the double-stranded iRNA agent Petition 870260049905, dated 05 / 25 / 2026, p. 26 / 728 13 / 335 has one or more 2'-F modifications in any position on the sense tape or antisense tape.

[0058] In some embodiments, the double-stranded iRNA agent has less than 20%, less than 15%, less than 10%, less than 5% non-natural nucleotides, or substantially no non-natural nucleotides. Examples of unnatural nucleotides include acyclic nucleotides, LNA, HNA, CeNA, 2'-O-methoxyalkyl (e.g., 2'-O-methoxymethyl, 2'-O-methoxyethyl, or 2-O-2-methoxypropanyl), 2'-O-allyl, 2'-C-allyl, 2'-fluorine, 2'-ON-methylacetamide (2-O-NMA), 2'-O-dimethylaminoethoxyethyl (2-O-DMAEOE), 2-O-aminopropyl (2-O-AP), 2'-ara-F, L-nucleoside modification (such as 2'-modified L-nucleoside, e.g., 2'-deoxy-L-nucleoside), abasic sugar of BNA, cyclic abasic, and open-chain alkyl.

[0059] In some embodiments, the double-stranded iRNA agent has more than 80%, more than 85%, more than 90%, more than 95%, or virtually 100% natural nucleotides. For the purpose of these embodiments, natural nucleotides may include those having 2'-OH, 2'-deoxy, and 2'-OMe.

[0060] In one embodiment, the double-stranded RNA agent comprises a sense strand and an antisense strand each having a length of 15–30 nucleotides; at least two phosphorothioate internucleotide bonds in the first five nucleotides in the antisense strand (counting from the 5' end); wherein the duplex region has between 19 and 25 base pairs (preferably 19, 20, 21, or 22); wherein the double-stranded RNA agent has less than 20%, less than 15%, less than 10%, less than 5% of unnatural nucleotides, or substantially no unnatural nucleotides.

[0061] In one embodiment, the double-stranded iRNA agent with Petition 870260049905, dated 05 / 25 / 2026, page 27 / 728 14 / 335 comprises a sense strand and an antisense strand, each having a length of 15-30 nucleotides; at least two phosphorothioate internucleotide bonds in the first five nucleotides in the antisense strand (counting from the 5' end); wherein the duplex region has between 19 and 25 base pairs (preferably 19, 20, 21, or 22); wherein the double-stranded iRNA agent has more than 80%, more than 85%, more than 95%, or virtually 100% natural nucleotides, such as those having 2'-OH, 2-deoxy, or 2'-OMe.

[0062] Another aspect of the invention relates to a method of reducing the expression of a target gene in a cell, comprising contacting said cell with a double-stranded RNA agent comprising an antisense strand that is complementary to a target gene; a sense strand that is complementary to said antisense strand; and one or more lipophilic portions conjugated to one or more internal positions in at least one strand, optionally via a linker or carrier.

[0063] All the above embodiments relating to lipophilic moieties and their conjugation to the double-stranded RNA agent in the first aspect of the invention relating to the double-stranded RNA agent are suitable in this aspect of the invention relating to a method of reducing the expression of a target gene in a cell.

[0064] In one embodiment, the cell is an extrahepatic cell.

[0065] Another aspect of the invention relates to a method of reducing the expression of a target gene in an individual, comprising administering to the individual a double-stranded RNA agent comprising contact of said cell with a double-stranded RNA agent comprising an antisense strand that is complementary to a target gene; a sense strand that is complementary to said antisense strand; and one or more lipophilic portions conjugated to one or more Petition 870260049905, dated 05 / 25 / 2026, p. 28 / 728 15 / 335 internal positions on at least one tape, optionally via a binder or vehicle.

[0066] All the above embodiments relating to lipophilic moieties and their conjugation to the double-stranded RNA agent in the first aspect of the invention relating to the double-stranded RNA agent are suitable in this aspect of the invention relating to a method of reducing the expression of a target gene in an individual.

[0067] In some embodiments, the double-stranded iRNA agent is administered extrahepatically.

[0068] In one embodiment, the double-stranded RNA agent is administered intrathecally. By intrathecal administration of the double-stranded RNA agent, the method can reduce the expression of a target gene in a brain or spinal tissue, for example, cortex, cerebellum, cervical spine, lumbar spine, and thoracic spine.

[0069] In some modalities, exemplary target genes are APP, ATXN2, C9orf72, TARDBP, MAPT (Tau), HTT, SNCA, FUS, ATXN3, ATXN1, SCA1, SCA7, SCA8, MeCP2, PRNP, SOD1, DMPK, and TTR. To reduce the expression of these target genes in the individual, the double-stranded RNA agent can be administered intravitreally. By intravitreal administration of the double-stranded RNA agent, the method can reduce the expression of the target gene in ocular tissue.

[0070] Another aspect of the invention relates to a method of treating an individual having a CNS dysfunction, comprising administering to the individual a therapeutically effective amount of a double-stranded RNAi agent, thereby treating the individual. The double-stranded RNAi agent comprises an antisense strand that is complementary to a target gene; a sense strand that is complementary to said antisense strand; and one or more lipophilic portions conjugated to one or more internal positions in at least Petition 870260049905, dated 05 / 25 / 2026, p. 29 / 728 16 / 335 a tape, optionally via a binder or vehicle.

[0071] All the above embodiments relating to lipophilic moieties and their conjugation to the double-stranded RNA agent in the first aspect of the invention relating to the double-stranded RNA agent are suitable in this aspect of the invention relating to a method of treating an individual having a CNS dysfunction. Exemplary CNS dysfunctions that can be treated by the method of the invention include Alzheimer's, amyotrophic lateral sclerosis (ALS), frontotemporal dementia, Huntington's, Parkinson's, spinocerebellar, prion and Lafora. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 is a diagram showing ligands, such as lipophilic moieties, that are conjugated to siRNAs at internal sense or antisense strand positions (i.e., somewhere within the siRNA sequence).

[0073] Figure 2 is a diagram showing ligands, as lipophilic moieties, that are conjugated to siRNAs via ligands or carriers at the 3' and / or 5' ends of the sense or antisense strand.

[0074] Figure 3 is a diagram showing ligands, as lipophilic moieties, that are conjugated to siRNAs via biocleavable ligands.

[0075] Figure 4 is a graph showing the results of silencing the beta-catenin gene (ocular CTNNB1) by intravitreal injection of several exemplary siRNA conjugates in mice.

[0076] Figure 5 is a graph showing the results of SOD1 mRNA silencing by a single intrathecal injection of several exemplary siRNA conjugates into the cortex of Sprague Dawley mice.

[0077] Figure 6 is a graph showing the results of the silenciPetition 870260049905, dated 05 / 25 / 2026, page 30 / 728 17 / 335 SoD1 mRNA increase by a single intrathecal injection of several exemplary siRNA conjugates in the cerebellum of Sprague Dawley rats.

[0078] Figure 7 is a graph showing the results of SOD1 mRNA silencing by a single intrathecal injection of several exemplary siRNA conjugates in the cervical spine of Sprague Dawley rats.

[0079] Figure 8 is a graph showing the results of SOD1 mRNA silencing by a single intrathecal injection of several exemplary siRNA conjugates in the lumbar spine of Sprague Dawley rats.

[0080] Figure 9 is a graph showing the results of SOD1 mRNA silencing by a single intrathecal injection of several exemplary siRNA conjugates in the Thoracic Spine of Sprague Dawley Rats.

[0081] Figure 10 shows the results of free uptake of primary cine hepatocytes (PCH) (without transfection agent) into cells incubated with an F12 siRNA, modified by conjugation of a lipophilic portion (C16) at each position of the antisense and sense strands, at concentrations of 2.5 and 250 nM by measuring F12 mRNA levels after 24 hours using RT-qPCR.

[0082] Figure 11 shows the results of uptake of free primary cine hepatocytes (PCH) (without transfection agent) into cells incubated with an F12 siRNA, modified by conjugation of a lipophilic portion (C16) at each position of the antisense and sense strands, at concentrations of 2.5 and 250 nM by measuring F12 mRNA levels after 24 hours using RT-qPCR.

[0083] Figure 12 shows the relative hydrophobicity results for each position of the antisense and sense strands of a siRNA duplex, modified by conjugation of a lipophilic portion (C16) in Petition 870260049905, dated 05 / 25 / 2026, p. 31 / 728 18 / 335 each position of the antisense and sense strands, determined by measuring the unbound portion using an electrophoretic mobility shift assay after each siRNA conjugate has been incubated with human serum albumin.

[0084] Figures 13A-13C show that durable SOD1 mRNA silencing is seen in all brain and spinal cord regions tested. Figure 13A shows the results of SOD1 mRNA silencing by a single intrathecal injection of several exemplary siRNA conjugates in rats in the lumbar spine, thoracic spine, and cervical spine regions, respectively. Figure 13B is a diagram showing various tissues tested in the rat CNS. Figure 13C shows the results of SOD1 mRNA silencing by a single intrathecal injection of several exemplary siRNA conjugates in rats in the cerebellum, frontal cortex, and remaining brain regions, respectively.

[0085] Figures 14A-14B show the results of β-catenin silencing after a single intrathecal dose. Figure 14A shows the results of β-catenin silencing of several exemplary siRNA conjugates in rats in the lumbar spine, thoracic spine, and cervical spine regions, respectively. Figure 14B shows the results of β-catenin silencing of several exemplary siRNA conjugates in rats in the cerebellum, frontal cortex, and remaining brain regions, respectively.

[0086] Figures 15A-15C show the results of SOD1 silencing after a single intrathecal dosing of exemplary siRNA duplexes in rats, indicating higher drug levels and robust silencing observed in the brain with SOD1 siRNA conjugate. Figure 15A shows the levels of conjugated siRNA in the CSF, compared to the levels of unconjugated siRNA. Figure 15B shows the levels of conjugated siRNA in the brain, compared to the levels of unconjugated siRNA. Petition 870260049905, dated 05 / 25 / 2026, p. 32 / 728 19 / 335 Comparison with unconjugated siRNA levels. Figure 15C shows conjugated siRNA levels in the cerebellum compared with unconjugated siRNA levels and control siRNA levels.

[0087] Figures 16A-16B show the results of SOD1 silencing with different chemical modifications at various doses. Figure 16A shows the results of SOD1 silencing in rats in the lumbar spine, thoracic spine, and cervical spine regions, respectively. Figure 16B shows the results of SOD1 silencing in rats in the cerebellum, frontal cortex, and remaining brain regions, respectively.

[0088] Figure 17 shows the results of βcatenin siRNA levels after a single intrathecal (IT) measurement of an exemplary siRNA duplex in various non-human primate (NHP) regions on Day 31.

[0089] Figure 18 shows the results of robust gene silencing of β-catenin mRNA in various tissues, on Day 31.

[0090] Figure 19 shows images illustrating siRNAs distributed throughout the CNS in NHP, after a single IT measurement.

[0091] Figure 20 shows images illustrating siRNA conjugates located in neurons after a single IT measurement. MAP2 is a neuronal marker.

[0092] Figure 21 shows images illustrating siRNA conjugates located in microglia after a single IT assay. Iba1 is a microglia marker.

[0093] Figure 22 shows images illustrating siRNA conjugates located in astrocytes, after a single IT measurement.

[0094] Figure 23 shows a graph comparing gene silencing activity observed in rats and NHP at dose-escalating compartment levels.

[0095] Figure 24 shows the results of mRNA levels of Petition 870260049905, dated 05 / 25 / 2026, page 33 / 728 20 / 335 TTR in the eye of mice on Day 14 after administration of several exemplary siRNA duplexes shown in Table 7, at a dosage of 3 pg or 7.5 pg.

[0096] Figure 25 shows the results of TTR mRNA levels in the eye of mice on Day 14 after administration of several exemplary siRNA duplexes shown in Table 7, at a dosage of 7.5 pg.

[0097] Figure 26 shows the results of TTR mRNA levels in the eye of mice on Day 14 after intravitreal administration of several exemplary siRNA duplexes shown in Table 7, at a dosage of 7.5 pg. DETAILED DESCRIPTION

[0098] The inventors discovered, inter alia, that conjugating a lipophilic moiety to one or more internal positions in at least one strand of the double-stranded iRNA agent provides surprisingly good results for intravitreal delivery and in vivo intrathecal delivery of double-stranded iRNAs, resulting in efficient entry into CNS and ocular tissues and efficient internalization into cells of the CNS and ocular system.

[0099] One aspect of the invention provides a double-stranded RNA agent comprising: an antisense strand that is complementary to a target gene; a sense strand that is complementary to said antisense strand; and one or more lipophilic portions conjugated to one or more internal positions in at least one strand, optionally via a linker or carrier.

[00100] The term lipophilic or lipophilic portion broadly refers to any compound or chemical portion having an affinity for lipids. One way to characterize the lipophilicity of the lipophilic portion is by the octanol-water partition coefficient, logKow, where Kow is the ratio between the concentration of a chemical in the octanol phase and its concentration in the water phase. Petition 870260049905, dated 05 / 25 / 2026, p. 34 / 728 21 / 335 traction in the aqueous phase of a two-phase system in equilibrium. The octanol-water partition coefficient is a property of a substance measured in the laboratory. However, it can also be predicted using coefficients assigned to the structural components of a chemical that are calculated using first-principles or empirical methods (see, for example, Tetko et al., J. Chem. Inf. Comput. Sci. 41: 1407-21 (2001), which is incorporated here by reference in its entirety). It provides a thermodynamic measure of the substance's tendency to prefer a non-aqueous or oily medium over water (i.e., its hydrophilic / lipophilic equilibrium). In principle, a chemical substance has lipophilic character when its logKow exceeds 0. Typically, the moiety has a logKow exceeding 1, exceeding 1.5, exceeding 2, exceeding 3, exceeding 4, exceeding 5, or exceeding 10. For example, the logKow of 6-aminohexanol is predicted to be approximately 0.7.Using the same method, the logKow of cholesteryl N-(hexan-6-ol)carbamate is predicted to be 10.7.

[00101] The lipophilicity of a molecule can change with respect to the functional group it carries. For example, the addition of a hydroxyl group or an amine group to the end of a lipophilic portion can increase or decrease the value of the partition coefficient (e.g., logKow) of the lipophilic portion.

[00102] Alternatively, the hydrophobicity of the double-stranded RNA agent, conjugated to one or more lipophilic moieties, can be measured by its protein binding characteristics. For example, the unbound moiety in the plasma protein binding assay of the double-stranded RNA agent can be determined to correlate positively with the relative hydrophobicity of the double-stranded RNA agent, which can correlate positively with the silencing activity of the double-stranded RNA agent.

[00103] In one embodiment, the protein binding assay Petition 870260049905, dated 05 / 25 / 2026, page 35 / 728 22 / 335 plasma determined is an electrophoretic mobility shift assay (EMSA) using human serum albumin protein. An exemplary protocol of this binding assay is illustrated in detail in Example 14. The hydrophobicity of the double-stranded iRNA agent, measured by the unbound siRNA portion in the binding assay, exceeds 0.15, exceeds 0.2, exceeds 0.25, exceeds 0.3, exceeds 0.35, exceeds 0.4, exceeds 0.45, or exceeds 0.5 for enhanced in vivo siRNA delivery.

[00104] Accordingly, conjugation of the lipophilic portions to the internal position(s) of the double-stranded iRNA agent provides optimal hydrophobicity for enhanced in vivo delivery of siRNA.

[00105] In certain embodiments, the lipophilic portion is an aliphatic compound, cyclic such as alicyclic or polycyclic such as polyalicyclic, such as a steroid (e.g., sterol) or a linear or branched aliphatic hydrocarbon. The lipophilic portion may generally comprise a hydrocarbon chain, which may be cyclic or acyclic. The hydrocarbon chain may comprise several substituents and / or one or more heteroatoms, such as an oxygen or nitrogen atom. Such lipophilic aliphatic moieties include, without limitation, saturated or unsaturated C4-C30 hydrocarbons (e.g., Oθ-Oιβ hydrocarbons), saturated or unsaturated fatty acids, waxes (e.g., monohydric alcohol esters of fatty acids and fatty diamides), terpenes (e.g., C10 terpenes, C15 sesquiterpenes, C20 diterpenes, C30 triterpenes and C40 tetraterpenes) and other polyalicyclic hydrocarbons.For example, the lipophilic portion may contain a C4-C30 hydrocarbon chain (e.g., C4-C30 alkyl or alkenyl). In some embodiments, the lipophilic portion contains a saturated or unsaturated Ce-Cie hydrocarbon chain (e.g., a linear Ce-Cie alkyl or alkenyl). In one embodiment, the lipophilic portion contains a saturated C16 hydrocarbon chain. Petition 870260049905, dated 05 / 25 / 2026, p. 36 / 728 23 / 335 or unsaturated (for example, a linear Cw alkyl or alkenyl).

[00106] The lipophilic moiety can be attached to the iRNA agent by any method known in the art, including through a functional group already present in the lipophilic moiety or introduced into the iRNA agent, such as a hydroxyl group (e.g., -CO-CH2OH). Functional groups already present in the lipophilic moiety or introduced into the iRNA agent include, but are not limited to, hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide, and alkyne.

[00107] Conjugation of the RNA agent and the lipophilic moiety can occur, for example, through the formation of a carboxylic ether or carbamoyl linkage between the hydroxyl and an alkyl group R-, an alkanol group RCO-, or a substituted carbamoyl group RNHCO-. The alkyl group R can be cyclic (e.g., cyclohexyl) or acyclic (e.g., linear or branched chain; and saturated or unsaturated). The alkyl group R can be a butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, or octadecyl group, or similar.

[00108] In some embodiments, the lipophilic portion is conjugated to the double-stranded RNA agent via a linker containing an ether, thioether, urea, carbonate, amine, amide, maleimidathioether, disulfide, phosphodiester, sulfonamide linkage, a product of a click reaction (e.g., a triazole from the azide-alkyne cycloaddition) or carbamate.

[00109] In another embodiment, the lipophilic portion is a steroid, such as a sterol. Steroids are polycyclic compounds containing a perhydro-1,2-cyclopentanophenanthrene ring system. Steroids include, without limitation, bile acids (e.g., cholic acid, deoxycholic acid, and dehydrocholic acid), cortisone, digoxigenin, testosterone, cholesterol, and cationic steroids such as cortisone. A cholesterol derivative refers to a compound derived from cholesterol. Petition 870260049905, dated 05 / 25 / 2026, page 37 / 728 24 / 335 for example by substitution, addition or removal of substituents.

[00110] In another embodiment, the lipophilic moiety is an aromatic moiety. In this context, the term aromatic refers broadly to mono- and polyaromatic hydrocarbons. Aromatic groups include, without limitation, Ce-Cu aryl moieties comprising one to three aromatic rings, which may be optionally substituted; aralkyl or arylalkyl groups comprising an aryl group covalently bonded to an alkyl group, either of which may be independently optionally substituted or unsubstituted; and heteroaryl groups. As used herein, the term heteroaryl refers to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14π shared electrons in a cyclic matrix, and having, in addition to carbon atoms, between one and about three heteroatoms selected from the group consisting of nitrogen (N), oxygen (O), and sulfur (S).

[00111] As used herein, a substituted alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic group is one having between one and about four, preferably between one and about three, more preferably one or two, substituents other than hydrogen. Suitable substituents include, without limitation, halo, hydroxy, nitro, haloalkyl, alkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano and ureido groups.

[00112] In some embodiments, the lipophilic moiety is an aralkyl group, for example, a 2-arylpropanoyl moiety. The structural features of the aralkyl group are selected such that the lipophilic moiety binds to at least one protein in vivo. In certain embodiments, the structural features of the aralkyl group are selected Petition 870260049905, dated 05 / 25 / 2026, p. 38 / 728 25 / 335 such that the lipophilic portion binds to serum, vascular, or cellular proteins. In certain embodiments, the structural features of the aralkyl group promote binding to albumin, an immunoglobulin, a lipoprotein, α-2-macroglobulin, or α-1-glycoprotein.

[00113] In certain embodiments, the ligand is naproxen or a structural derivative of naproxen. Procedures for the synthesis of naproxen can be found in U.S. Pat. No. 3,904,682 and U.S. Pat. No. 4,009,197, which are hereby incorporated by reference in their entirety. Naproxen has the chemical name (S)-6-methoxy-α-methyl-2-naphthaleneacetic acid and the structure is

[00114] In certain embodiments, the ligand is ibuprofen or a structural derivative of ibuprofen. Procedures for the synthesis of ibuprofen can be found in U.S. Pat. No. 3,228,831, which are hereby incorporated by reference in their entirety. The structure of ibuprofen is

[00115] Additional exemplary aralkyl groups are illustrated in U.S. Patent No. 7,626,014, which is incorporated herein by reference in its entirety.

[00116] In another embodiment, suitable lipophilic moieties include lipid, cholesterol, retinoic acid, cholic acid, adamantanoacetic acid, 1-pyrenebutyric acid, dihydrotestosterone, 1,3-bisO(hexadecyl)glycerol, geranyloxyhexanol, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, ibuprofen, naproxen, dimethoxytritil or phenoxazine. Petition 870260049905, dated 05 / 25 / 2026, page 39 / 728 26 / 335

[00117] In some embodiments, the lipophilic portion is a C6-C30 acid (e.g., hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, octadecanoic acid, oleic acid, linoleic acid, arachidonic acid, cis4,7,10,13,16,19-docosahexanoic acid, vitamin A, vitamin E, cholesterol, etc.) or a C6-C30 alcohol (e.g., hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol, tridecanol, tetradecanol, pentadecanol, hexadecanol, heptadecanol, octadecanol, oleyl alcohol, alcohol linoleic acid, arachidonic alcohol, c / s-4,7,10,13,16,19-docosahexaenolic acid, retinol, vitamin E, cholesterol, etc.).

[00118] In certain embodiments, more than one lipophilic moiety may be incorporated into the double-stranded RNA agent, particularly when the lipophilic moiety has low lipophilicity or hydrophobicity. In one embodiment, two or more lipophilic moieties are incorporated into the same strand of the double-stranded RNA agent. In one embodiment, each strand of the double-stranded RNA agent has one or more lipophilic moieties incorporated. In one embodiment, two or more lipophilic moieties are incorporated at the same position (i.e., the same nucleobase, same sugar moiety, or same internucleosidic bond) of the double-stranded RNA agent. This can be achieved by, for example, conjugation of the two or more lipophilic portions via a vehicle and / or conjugation of the two or more lipophilic portions via a branched ligand and / or conjugation of the two or more lipophilic portions via one or more ligands, with one or more ligands connecting the lipophilic portions consecutively.

[00119] The lipophilic portion can be conjugated to the iRNA agent via direct attachment to the ribosugar of the iRNA agent. Alternatively, the lipophilic portion can be conjugated to the agent of Petition 870260049905, dated 05 / 25 / 2026, p. 40 / 728 27 / 335 double-stranded iRNA via a linker or a vehicle.

[00120] In certain embodiments, the lipophilic portion can be conjugated to the iRNA agent via one or more ligands (ties).

[00121] In one embodiment, the lipophilic portion is conjugated to the double-stranded RNA agent via a linker containing an ether, thioether, urea, carbonate, amine, amide, maleimide-thioether, disulfide, phosphodiester, sulfonamide, a product of a click reaction (e.g., a triazole of the azide-alkyne cycloaddition), or carbamate linkage. Some exemplary linkages are illustrated in Figure 1, Examples 2, 3, 5, 6, and 7. Binders / Ties

[00122] The ligands / ties are connected to the lipophilic portion at a tether attachment point (TAP). The ligands / ties may include any C1-C100 carbon-containing portion (e.g., C1-C75, C1-C50, C1-C20, C1-C10; C1, C2, C3, C4, C5, C6, C7, C8, Cg, or C10) and may have at least one nitrogen atom. In certain embodiments, the nitrogen atom forms part of a terminal amino or amide group (NHC(O)-) in the ligand / ties, which may serve as a connection point for the lipophilic portion. Non-limited examples of ligands / ties (underlined) include TAP-(CH2)nNH-; TAPC(O)(CH2)nNH-; TAP-NR(CH2)nNH-, TAP-C(O)-(CH2)nC(O)-; TAPC(O)-(CH2)nC(O)O-; TAP-C(O)-O-; TAP-C(O)-(CH2)n-NH-C(O)-; TAPC(O)-(CH2)n-; TAP-C(O)-NH-; TAP-C(O)-; TAP-(CH2)nC(O)-; TAP(CH2)nC(O)O-; TAP-(CH2)n-; or TAP-(CH2)n-NH-C(O)-; wherein n is 1-20 (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) and R is C1-C1 alkyl. Preferably, n is 5, 6 or 11.In other embodiments, the nitrogen may form part of a terminal oxyamino group, for example, -ONH2, or a hydrazino group, NHNH2. The linker / bracket may be optionally substituted, for example, by hydroxy, alkoxy, per-haloalkyl and / or optionally inserted. Petition 870260049905, dated 05 / 25 / 2026, page 41 / 728 28 / 335 with one or more additional heteroatoms, for example, N, O or S. Preferred tethered ligands may include, for example, TAP2(CH2)nNH(LIGAND); TAP-C(Q)(CH2)nNH(LIGAND); TAPNR(CH2)nNH(LIGAND); TAP-(CH2)nONH(LIGAND); TAPC(O)(CH2)nONH(LIGAND); TAP-NR(CH2)nONH(LIGAND); TAP(CH2)nNHNH2(LIGAND); TAP-C(O)(CH2)nNHNH2(LIGAND); TAPNR(CH2)nNHNH2(LIGAND); TAP-C(O)-(CH2)nC(O)(LIGAND); TAP-C(Q)-(CH2)nC(Q)O(LIGAND); TAP-C(O)-O(LIGAND); TAPC(Q)-(CH2)n-NH-C(Q)(LIGAND); TAP-C(O)-(CH2)n(LIGAND); TAPC(Q)-NH(LIGANDQ); TAP-C(O)(LIGAND); TAP-(CH2)nC(O)(LIGAND); TAP-(CH2)nC(O)O(LIGAND); TAP-(CH2)n(LIGAND); or TAP-(CH2)n-NH-C(O)(LIGAND). In some embodiments, amino-terminated ligands / ties (e.g., NH2, ONH2, NH2NH2) can form an imino bond (i.e., C=N) with the ligand. In some embodiments, amino-terminated ligands / ties (e.g., NH2, ONH2, NH2NH2) can be acylated, for example, with C(O)CF3.

[00123] In some embodiments, the linker / tie may terminate with a mercapto group (i.e., SH) or an olefin (e.g., CH=CH2). For example, the tie may be TAP-(CH2)n-SH, TAPC(O)(CH2)nSH, TAP-(CH2)n-(CH=CH2) or TAP-C(O)(CH2)n(CH=CH2), where n may be as described elsewhere. The tie may optionally be replaced, for example, by hydroxy, alkoxy, perhaloalkyl and / or optionally inserted with one or more additional heteroatoms, for example, N, O or S. The double bond may be cis or trans or E or Z.

[00124] In other embodiments, the linker / tie may include an electrophilic moiety, preferably at the terminal position of the linker / tie. Exemplary electrophilic moieties include, for example, an aldehyde, alkyl halide, mesylate, tosylate, nosylate, or brosylate. Petition 870260049905, dated 05 / 25 / 2026, p. 42 / 728 29 / 335 or an activated carboxylic acid ester, for example, an NHS ester or a pentafluorophenyl ester. Preferred ligands / ties (underlined) include TAP-(CH2)nCHQ; TAP-C(O)(CH2)nCHO; or TAPNR(CH2)nCHO, where n is 1-6 and R is C1-C6 alkyl; or TAP2(CH2)nC(O)ONHS; TAP-C(O)(CH2)nC(O)ONHS; or TAP-NR(CH2)nC(Q)QNHS, where n is 1-6 and R is C1-C6 alkyl; TAP(CH2)nC(Q)QC6F5; TAP-C(O)(CH2)nC(O)OC6F5; or TAP-NR(CH2)nC(Q) QCeFs, where n is 1-11 and R is Ci-Ce alkyl; or -(CH2)nCH2LG; TAP-C(Q)(CH2)nCH2LG; or TAP-NR(CH2)nCH2LG, in which n may be as described elsewhere and R is Ci-Ce alkyl (LG may be a labile group, for example, halide, mesylate, tosylate, nosylate, brosylate). The latch may be achieved by coupling a nucleophilic group of a ligand, for example, a thiol or amino group, with an electrophilic group in the latch.

[00125] In other embodiments it may be desirable that the monomer include a phthalimide (K) group at the terminal position of the ligand. te / amarra. κ

[00126] In other embodiments, other protected amino groups may be in the terminal position of the linker / tie, for example, allo, monomethoxy trityl (MMT), trifluoroacetyl, Fmoc or aryl sulfonyl (for example, the aryl moiety may be ortho-nitrophenyl or ortho, paradinitrophenyl).

[00127] Any of the linkers / bonds described here may additionally include one or more additional linking groups, for example, -Q-(CH2)n-, -(CH2)n-SS-, -(CH2)n- or -(CH=CH)-. Cleavage ties / tie-downs

[00128] In some embodiments, at least one of the linkers / ties may be a redox cleavable linker, a linker cleavable by Petition 870260049905, dated 05 / 25 / 2026, p. 43 / 728 30 / 335 acid, an esterase-cleavable ligand, a phosphatase-cleavable ligand, or a peptidase-cleavable ligand.

[00129] In one embodiment, at least one of the linkers / ties can be a reductively cleavable linker (e.g., a disulfide group).

[00130] In one embodiment, at least one of the linkers / ties may be an acid-cleavable linker (e.g., a hydrazone group, an ester group, an acetal group, or a ketal group).

[00131] In one embodiment, at least one of the linkers / ties may be a linker that is cleavable by esterase (e.g., an ester group).

[00132] In one embodiment, at least one of the linkers / ties may be a phosphatase-cleavable linker (e.g., a phosphate group).

[00133] In one embodiment, at least one of the linkers / ties may be a peptidase-cleavable linker (e.g., a peptide linkage).

[00134] Cleavable linkage groups are susceptible to cleavage agents, for example, pH, redox potential, or the presence of degradation molecules. Generally, cleavage agents are more prevalent or found at higher levels or activities within cells than in serum or blood. Examples of such degradation agents include: redox agents that are selected for particular substrates or that have no substrate specificity, including, for example, oxidative or reductive enzymes or reducing agents such as mercaptans, present in cells, which can degrade a redox cleavable linkage group by reduction; esterases; endosomes or agents that can create an acidic environment, for example, those that result in a pH of five or lower; enzymes that can hydrolyze or degrade a cleavable linkage group. Petition 870260049905, dated 05 / 25 / 2026, page 44 / 728 31 / 335 by acting as a general acid, peptidases (which can be substrate-specific) and phosphatases.

[00135] A cleavable linking group, such as a disulfide bond, may be susceptible to pH. The pH of human serum is 7.4, while the average intracellular pH is slightly lower, ranging from about 7.1-7.3. Endosomes have a more acidic pH, in the range of 5.5-6.0, and lysosomes have an even more acidic pH around 5.0. Some ligands will have a linking group that is cleaved at a preferred pH, thereby releasing the iRNA agent from a ligand (e.g., a targeting or cell-permeable ligand, such as cholesterol) within the cell or into the desired cell compartment.

[00136] A chemical junction (e.g., a linking group) that links a ligand to an iRNA agent may include a disulfide bond. When the iRNA / ligand complex is absorbed into the cell by endocytosis, the acidic environment of the endosome will cause the disulfide bond to be cleaved, thereby releasing the iRNA agent from the ligand (Quintana et al., Pharm Res. 19: 1310-1316, 2002; Patri et al., Curr. Opin. Curr. Biot. 6: 466-471, 2002). The ligand may be a targeting ligand or a second therapeutic agent that may complement the therapeutic effects of the iRNA agent.

[00137] A tether may include a linking group that is cleavable by a particular enzyme. The type of linking group incorporated into a tether may depend on the cell being targeted by the iRNA agent. For example, an iRNA agent targeting an mRNA in liver cells may be conjugated to a tether that includes an ester group. Liver cells are rich in esterase, and therefore the tether will be cleaved more efficiently in liver cells than in cell types that are not rich in esterases. Cleavage of the tether releases the Petition 870260049905, dated 05 / 25 / 2026, page 45 / 728 32 / 335 an iRNA agent is a ligand that is attached to the distal end of the tether, thereby potentially intensifying the silencing activity of the iRNA agent. Other cell types rich in esterases include lung cells, renal cortex, and testes.

[00138] Tethers containing peptide linkages can be conjugated to RNA agents targeting peptidase-rich cell types, such as liver cells and synoviocytes. For example, an RNA agent targeting synoviocytes, such as for the treatment of an inflammatory disease (e.g., rheumatoid arthritis), can be conjugated to a tether containing a peptide linkage.

[00139] In general, the suitability of a candidate cleavable link group can be assessed by testing the ability of a degradation agent (or condition) to cleave the candidate link group. It will also be desirable to test the candidate cleavable link group for its ability to resist cleavage in blood or when in contact with other non-target tissue, for example, tissue to which the iRNA agent would be exposed when administered to an individual. Thus, the relative susceptibility to cleavage can be determined between a first and a second condition, where the first is selected to be indicative of cleavage in a target cell and the second is selected to be indicative of cleavage in other tissues or biological fluids, for example, blood or serum. Assessments can be carried out in cell-free systems, in cells, in cell culture, in organ or tissue culture, or in whole animals.It may be useful to perform initial assessments under cell-free or culture-free conditions and confirm by further assessments in whole animals. In preferred embodiments, useful candidate compounds are cleaved at least 2, 4, 10, or 100 times more rapidly in the cell (or under in vitro conditions selected to mimic intracellular conditions) compared with blood or serum (or under in vitro conditions). Petition 870260049905, dated 05 / 25 / 2026, page 46 / 728 33 / 335 selected to mimic extracellular conditions). Redox Cleavageable Link Groups

[00140] A class of cleavable linking groups are redox-cleavable linking groups that are cleaved upon reduction or oxidation. An example of a reductively cleavable linking group is a disulfide linking group (-SS-). To determine whether a candidate cleavable linking group is a suitable reductively cleavable linking group or, for example, is suitable for use with a particular RNAi moiety and particular targeting agent, methods described herein may be considered. For example, a candidate may be evaluated by incubation with dithiothreitol (DTT), or another reducing agent using reagents known in the art, which mimic the cleavage rate that would be observed in a cell, for example, a target cell. Candidates may also be evaluated under conditions that are selected to mimic blood or serum conditions. In a preferred embodiment, candidate compounds are cleaved by a maximum of 10% in blood.In preferred embodiments, useful candidate compounds are degraded at least 2, 4, 10, or 100 times more rapidly in the cell (or under in vitro conditions selected to mimic intracellular conditions) compared to blood (or under in vitro conditions selected to mimic extracellular conditions). The cleavage rate of candidate compounds can be determined using standard enzyme kinetic assays under conditions chosen to mimic intracellular media and compared with conditions chosen to mimic extracellular media. Phosphate-Based Cleavable Linking Groups

[00141] Cleavable phosphate-based linkage groups are cleaved by agents that degrade or hydrolyze the phosphate group. An example of an agent that cleaves phosphate groups in cells is enzyme Petition 870260049905, dated 05 / 25 / 2026, page 47 / 728 34 / 335 but such as phosphatases in cells. Examples of phosphate-based linking groups are -OP(O)(ORk)-O-, -OP(S)(ORk)-O-, -OP(S)(SRk)-O-, -SP(O)(ORk)-O-, -OP(O)(ORk)-S-, -SP(O)(ORk)-S-, OP(S)(ORk)-S-, -SP(S)(ORk)-O-, -OP(O)(Rk)-O-, -OP(S)(Rk)-O-, SP(O)(Rk)-O-, -SP(S)(Rk)-O-, -SP(O)(Rk)-S-, -OP(S)(Rk)-S-. Preferred modalities are -OP(O)(OH)-O-, -OP(S)(OH)-O-, -OP(S)(SH)-O-, -SP(O)(OH)-O-, -OP(O)(OH)-S-, -SP(O)(OH)-S-, -OP(S)(OH)-S-, -SP(S)(OH)-O-, -OP(O)(H)-O-, -OP(S)(H)-O-, -SP(O)(H)-O-, -SP(S)(H)-O-, -SP(O)(H)-S-, -OP(S)(H)-S-. A preferred modality is -OP(O)(OH)-O-. These candidates can be evaluated using methods analogous to those described above. Acid-Clevitable Linkage Groups

[00142] Acid-cleavable linking groups are linking groups that are cleaved under acidic conditions. In preferred embodiments, acid-cleavable linking groups are cleaved in an acidic environment with a pH of about 6.5 or lower (e.g., about 6.0, 5.5, 5.0 or lower) or by agents such as enzymes that can act as a general acid. In a cell, specific low-pH organelles, such as endosomes and lysosomes, can provide a cleavage environment for acid-cleavable linking groups. Examples of acid-cleavable linking groups include, but are not limited to, hydrazones, ketals, acetals, esters, and amino acid esters. Acid-cleavable groups may have the general formula -C=NN-, C(O)O, or -OC(O). A preferred embodiment is when the carbon attached to the oxygen of the ester (the alkoxy group) is an aryl group, substituted alkyl group, or tertiary alkyl group such as dimethyl pentyl or t-butyl.These candidates can be evaluated using methods similar to those described above. Ester-Based Linking Groups

[00143] Ester-based cleavable linking groups are cleaved Petition 870260049905, dated 05 / 25 / 2026, p. 48 / 728 35 / 335 by enzymes such as esterases and amidases in cells. Examples of ester-based cleavable linking groups include, but are not limited to, alkylene, alkenylene, and alkynylene esters. Ester-cleavable linking groups have the general formula -C(O)O- or OC(O)-. These candidates can be evaluated using methods analogous to those described above. Peptide-Based Cleavage Groups

[00144] Peptide-based cleavable linkages are cleaved by enzymes such as peptidases and proteases in cells. Peptide-based cleavable linkages are peptide bonds formed between amino acids to produce oligopeptides (e.g., dipeptides, tripeptides, etc.) and polypeptides. Peptide-based cleavable linkages do not include the amide group (-C(O)NH-). The amide group can be formed between any alkylene, alkenylene, or alkynylene. A peptide bond is a special type of amide bond formed between amino acids to produce peptides and proteins. The peptide-based cleavable linkage is generally limited to the peptide bond (i.e., the amide bond) formed between amino acids to produce peptides and proteins and does not include the entire amide functional group. Peptide-cleavable linking groups have the general formula -NHCHR1C(O)NHCHR2C(O)-, where R1 and R2 are the R groups of the two adjacent amino acids.These candidates can be evaluated using methods similar to those described above. Biocleavable binders / ties

[00145] Ligands may also include biocleavable ligands that are nucleotide and non-nucleotide ligands or combinations thereof that connect two parts of a molecule, for example, one or both strands of two individual siRNA molecules to generate a bis(siRNA). In some embodiments, the mere electrostatic or stacking interaction between two individual siRNAs may represent Petition 870260049905, dated 05 / 25 / 2026, page 49 / 728 36 / 335 a ligand. Non-nucleotide ligands include ligands or ligands derived from monosaccharides, disaccharides, oligosaccharides and their derivatives, aliphatic, alicyclic, heterocyclic and their combinations.

[00146] In some embodiments, at least one of the linkers (ties) is a biocleavable linker selected from the group consisting of DNA, RNA, disulfide, amide, functionalized monosaccharides or oligosaccharides of galactosamine, glucosamine, glucose, galactose and mannose and combinations thereof.

[00147] In one embodiment, the biocleavable carbohydrate linker may have 1 to 10 saccharide units, which have at least one anomeric linkage capable of connecting two siRNA units. When two or more saccharides are present, these units may be linked via 1-3, 1-4, or 1-6 sugar linkages or via alkyl chains.

[00148] Exemplary biocleavable ligands include: Petition 870260049905, dated 05 / 25 / 2026, p. 50 / 728 37 / 335 HO NHAc Petition 870260049905, dated 05 / 25 / 2026, p. 51 / 728 38 / 335

[00149] Additional exemplary biocleavable ligands are illustrated in Schemes 28-30.

[00150] Further discussion on biocleavable ligands can be found in CT application No. PCT / US18 / 14213, entitled Endosomal Cleavable Ligands, filed on January 18, 2018, the contents of which are incorporated herein by reference in their entirety. Vehicles

[00151] In certain embodiments, the lipophilic portion is conjugated to the iRNA agent via a carrier that replaces one or more nu Petition 870260049905, dated 05 / 25 / 2026, p. 52 / 728 39 / 335 cleotide(s).

[00152] The vehicle can be a cyclic group or an acyclic group. In one embodiment, the cyclic group is selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl, and decalin. In another embodiment, the acyclic group is a moiety based on a serinol skeleton or a diethanolamine skeleton.

[00153] In some embodiments, the carrier replaces one or more nucleotide(s) in the internal position(s) of the double-stranded iRNA agent.

[00154] In other embodiments, the carrier replaces the nucleotides at the terminal end of the sense strand or antisense strand. In one embodiment, the carrier replaces the terminal nucleotide at the 3' end of the sense strand, thus functioning as a terminal cap protecting the 3' end of the sense strand. In one embodiment, the carrier is a cyclic group having an amine, for example, the carrier may be pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuranyl or decalinyl.

[00155] A ribonucleotide subunit in which the ribose sugar of the subunit has been thus substituted is referred to here as a ribose substitution modification subunit (RRMS). The carrier may be a cyclic or acyclic moiety and include two attachment points to the backbone (e.g., hydroxyl groups) and a linker (e.g., the lipophilic moiety). The lipophilic moiety may be directly attached to the carrier or indirectly attached to the carrier by an intervening linker / tie, as described above. Petition 870260049905, dated 05 / 25 / 2026, page 53 / 728 40 / 335 w w

[00156] The ligand-conjugated monomer subunit can be the 5' or 3' terminal subunit of the iRNA molecule, that is, one of the two W groups can be a hydroxyl group, and the other W group can be a chain of two or more unmodified or modified ribonucleotides. Alternatively, the ligand-conjugated monomer subunit can occupy an internal position, and both W groups can be one or more unmodified or modified ribonucleotides. More than one ligand-conjugated monomer subunit may be present in an iRNA agent. Sugar-Substitution Based Monomers, for example, Ligand-Conjugated (Cyclic) Monomers

[00157] Cyclic sugar-substituted base monomers, for example, monomers conjugated to a sugar-substituted base ligand, are also referred to here as RRMS monomer compounds. The carriers may have the general formula (LCM-2) provided below (In this structure, the preferred backbone attachment points may be chosen from R1 or R2; R3 or R4; or R9 and R10 if Y is CR9R10 (two positions are chosen to give two backbone attachment points, for example, R1 and R4 or R4 and R9)). Preferred backbone attachment points include R7; R5 or R6 when X is CH2. The carriers are described below as an entity, which may be incorporated into a ribbon. Thus it is understood that the structures Petition 870260049905, dated 05 / 25 / 2026, page 54 / 728 41 / 335 also encompasses situations where one (in the case of a terminal position) or two (in the case of an internal position) of the attachment points, for example, R1 or R2; R3 or R4; or R9 or R10 (when Y is CR9R10) are connected to the phosphate-containing skeleton, or modified phosphate, for example, sulfur. For example, one of the R groups mentioned above may be -CH2-, where one bond is connected to the carrier and one to a skeleton atom, for example, a bonding oxygen or a central phosphorus atom. (LCM-2) where: X is N(CO)R7, NR7 or CH2; Yé NR8, O, S, CR9R10; Z is CR11R12 or is absent; Each of R1, R2, R3, R4, R9 and R10 is, independently, H, ORa or (CH2)nORb, provided that at least two of R1, R2, R3, R4, R9 and R10 are ORa or (CH2)nORb; Each of R5, R6, R11 and R12 is independently a C1-C1 alkyl ligand optionally substituted by 1-3 R13 or C(O)NHR7; or R5 and R11 are together C3-C8 cycloalkyl optionally substituted by R14; R7 may be a linker, for example, R7 may be Rd, or R7 may be a linker indirectly tied to the vehicle, for example, through a tie portion, for example, Ci-C2o alkyl substituted by NRcRd; or Ci-C2o alkyl substituted by NHC(O)Rd; R8e H or Ci-Ce alkyl; R13 is hydroxy, C1-C4 alkoxy, or halo; R14 is NRCR7; Petition 870260049905, dated 05 / 25 / 2026, p. 55 / 728 42 / 335 R15 is C1-Ce alkyl optionally substituted by cyano or C2-C6 alkenyl; R16 is a C1-C10 alkyl group; R17 is a liquid or solid phase supporting reagent; L is -C(O)(CH2)qC(O)- or -C(O)(CH2)qS-; Ra is a protecting group, for example, CAra; (for example, a dimethoxytrityl group) or Si(X5')(X5)(X5') in which (X5), (X5) and (X5') are as described elsewhere. Rbé P(O)(O-)H, P(OR15)N(R16)2or L-R17; Rcé H or O-Ce alkyl; Rdé H or a ligand; Each Ar is independently aryl Ce-Cw optionally replaced by alkoxy C1-C4; n is 1-4; eq is 0-4.

[00158] Exemplary vehicles include those in which, for example, X is N(CO)R7 or NR7, Y is CR9R10, and Z is absent; or X is N(CO)R7 or NR7, Y is CR9R10, and Z is CR11R12; or X is N(CO)R7 or NR7, Y is NR8, and Z is CR11R12; or X is N(CO)R7 or NR7, Y is O, and Z is CR11R12; or X is CH2; Y is CR9R10; Z is CR11R12, and R5 and R11 together form C5 cycloalkyl (H, z = 2), or the indane ring system, for example, X is CH2; Y is CR9R10; Z is CR11R12, and R5 and R11 together form C5 cycloalkyl (H, z = 1).

[00159] In certain embodiments, the vehicle may be based on the pyrroline ring system or the 4-hydroxyproline ring system, for example, N(CO)R7 or NR7, Y is CR9R10, and Z is absent (D). OFG2 C4^-€^CH2OFG1( .Λ 2 . OFG1 is preferentially attached to a carbon CONNECTING Petition 870260049905, dated 05 / 25 / 2026, page 56 / 728 43 / 335 in the primary, for example, an exocyclic alkylene group, for example, a methylene group, connected to one of the carbons in the five-membered ring (-CH2OFG1 in D). OFG2 is preferably attached directly to one of the carbons in the five-membered ring (-OFG2 in D). For pyrroline-based vehicles, -CH2OFG1 can be attached to C-2 and OFG2 can be attached to C-3; or -CH2OFG1 can be attached to C-3 and OFG2 can be attached to C-4. In certain embodiments, CH2OFG1 and OFG2 can be geminally substituted by one of the carbons referenced above. For 3-hydroxyproline-based vehicles, -CH2OFG1 can be attached to C-2 and OFG2 can be attached to C-4. Pyrroline and 4-hydroxyproline-based monomers can therefore contain linkages (e.g., carbon-carbon linkages) in which rotation of the linkage is restricted around that particular linkage, for example, restriction resulting from the presence of a ring.Thus, CH2OFG1 and OFG2 can be cis / trans or trans with respect to each other in any of the pairings outlined above. Accordingly, all cis / trans isomers are expressly included. The monomers may also contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, individual enantiomers, individual diastereomers, and diastereomeric mixtures. All such isomeric forms of the monomers are expressly included (e.g., the centers carrying CH2OFG1 and OFG2 may both have the R configuration; or both have the S configuration; or one center may have the R configuration and the other center may have the S configuration and vice versa). The point of attachment to the mooring is preferably nitrogen. Preferred examples of vehicle D include the following: Petition 870260049905, dated 05 / 25 / 2026, page 57 / 728 44 / 335 GFO

[00160] In certain embodiments, the vehicle may be based on the piperidine ring system (E), for example, X is N(CO)R7 or NR7, Y is OFG2η---(CH2)nOFG / c2 N CONNECTING CR9R10e Z is CR11R12. e

[00161] OFG1 is preferably attached to a primary carbon, for example, an exocyclic alkylene group, for example, a methylene group (n=1) or an ethylene group (n=2), connected to one of the carbons in the six-membered ring [-(CH2)nOFG1 in E]. OFG2 is preferably attached directly to one of the carbons in the six-membered ring (-OFG2 in E). -(CH2)nOFG1 and OFG2 can be arranged in Petition 870260049905, dated 05 / 25 / 2026, page 58 / 728 45 / 335 a geminal arrangement in the ring, that is, both groups can be attached to the same carbon, for example, at C-2, C-3 or C-4. Alternatively, -(CH2)nOFG1 and OFG2 can be arranged in a vicinal arrangement in the ring, that is, both groups can be attached to adjacent ring carbon atoms, for example, (CH2)nOFG1 can be attached to C-2 and OFG2 can be attached to C-3; (CH2)nOFG1 can be attached to C-3 and OFG2 can be attached to C-2; (CH2)nOFG1 can be attached to C-3 and OFG2 can be attached to C-4; or (CH2)nOFG1 can be attached to C-4 and OFG2 can be attached to C-3. Piperidine-based monomers can therefore contain bonds (e.g., carbon-carbon bonds) in which bond rotation is restricted around that particular bond, for example, restriction resulting from the presence of a ring. Thus, -(CH2)nOFG1 and OFG2 can be cis or trans relative to each other in any of the pairings outlined above.Accordingly, all cis / trans isomers are expressly included. Monomers may also contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, individual enantiomers, individual diastereomers, and diastereomeric mixtures. All such isomeric forms of the monomers are expressly included (e.g., centers carrying CH2OFG1 and OFG2 may both have the R configuration; or both have the S configuration; or one center may have the R configuration and the other center may have the S configuration and vice versa). The point of attachment to the tie is preferably nitrogen.

[00162] In certain embodiments, the vehicle may be based on the piperazine ring system (F), for example, X is N(CO)R7 or NR7, Y is NR8, and Z is CR11R12, or on the morpholine ring system (G), for example, X is N(CO)R7 or NR7, Y is O, and Z is CR11R12. Petition 870260049905, dated 05 / 25 / 2026, page 59 / 728 46 / 335 I OFG2 N, / OFG2ch2ofg' ch2ofg CONNECTING The linker . OFG1 is preferably attached to a primary carbon, for example, an exocyclic alkylene group, for example, a methylene group, connected to one of the carbons in the six-membered ring (-CH2OFG1 in F or G). OFG2 is preferably attached directly to one of the carbons in the six-membered rings (-OFG2 in F or G). For both F and G, CH2OFG1 can be attached to C-2 and OFG2 can be attached to C-3; or vice versa. In certain embodiments, CH2OFG1 and OFG2 may be geminally substituted for one of the carbons referenced above. Piperazine and morpholine-based monomers may therefore contain linkages (e.g., carbon-carbon linkages) in which the rotation of the linkage is restricted around that particular linkage, for example, restriction resulting from the presence of a ring. Thus, CH2OFG1 and OFG2 can be cis or trans with respect to each other in any of the pairings outlined above. Accordingly, all cis / trans isomers are expressly included.Monomers may also contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, individual enantiomers, individual diastereomers, and diastereomeric mixtures. All such isomeric forms of monomers are expressly included (for example, the centers carrying CH2OFG1 and OFG2 may both have the R configuration; or both have the S configuration; or one center may have the R configuration and the other center may have the S configuration and vice versa). R' may be, for example, C1-C1 alkyl. Petition 870260049905, dated 05 / 25 / 2026, page 60 / 728 47 / 335 preferably CH3. The attachment point to the mooring is preferably nitrogen in both F and G.

[00163] In certain embodiments, the vehicle may be based on the decalin ring system, for example, X is CH2; Y is CR9R10; Z is CR11R12, and R5 and R11 together form C5 cycloalkyl (H, z = 2), or the indane ring system, for example, X is CH2; Y is CR9R10; Z is CR11R12 and R5 and R11 together form C5 cycloalkyl (H, z = 1). h. OFG1 is preferably attached to a primary carbon, for example, an exocyclic methylene group (n=1) or ethylene group (n=2) connected to one of C-2, C-3, C-4 or C-5 [(CH2)nOFG1 in H]. OFG2 is preferably attached directly to one of C-2, C-3, C-4 or C-5 (-OFG2 in H). -(CH2)nOFG1 and OFG2 can be arranged in a geminal manner in the ring, that is, both groups can be attached to the same carbon, for example, at C2, C-3, C-4 or C-5.Alternatively, -(CH2)nOFG1 and OFG2 can be arranged vicinally in the ring, that is, both groups can be attached to adjacent ring carbon atoms, for example, -(CFk^OFG1) can be attached to C-2 and OFG2 can be attached to C-3; -(CH2)nOFG1 can be attached to C-3 and OFG2 can be attached to C-2; -(CFk^OFG1) can be attached to C-3 and OFG2 can be attached to C-4; or -(CH2)nOFG1 can be attached to C-4 and OFG2 can be attached to C-3; -(CH2)nOFG1 can be attached to C-4 and OFG2 can be attached to C-5; or -(CH2)nOFG1 can be attached to C-5 and OFG2 can be attached to C-4. The monomers to Decalin or indane bases can therefore contain bonds (e.g., carbon-carbon bonds) in which bond rotation is restricted around that particular bond, for example, restriction resulting from the presence of a ring. Thus, -(CH2)nOFG1 and OFG2 can be cis or trans. Petition 870260049905, dated 05 / 25 / 2026, page 61 / 728 48 / 335 relative to each other in any of the pairings outlined above. Accordingly, all cis / trans isomers are expressly included. The monomers may also contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, individual enantiomers, individual diastereomers, and diastereomeric mixtures. All such isomeric forms of the monomers are expressly included (e.g., the centers carrying CH2OFG1 and OFG2 may both have the R configuration; or both have the S configuration; or one center may have the R configuration and the other center may have the S configuration and vice versa). In a preferred embodiment, the substituents at C-1 and C-6 are trans relative to each other. The point of attachment to the bond is preferably C-6 or C-7.

[00164] Other vehicles may include those based on 32GLO----Z 5----(CH^OFG, I By linking hydroxyproline (J). Thus, -(CFDnOFG1 and OFG2) can be cis or trans relative to each other. Accordingly, all cis / trans isomers are expressly included. The monomers may also contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, individual enantiomers, individual diastereomers, and diastereomeric mixtures. All such isomeric forms of the monomers are expressly included (e.g., the centers carrying CH2OFG1 and OFG2 may both have the R configuration; or both have the S configuration; or one center may have the R configuration and the other center may have the S configuration and vice versa). The point of attachment to the linkage is preferably nitrogen. Petition 870260049905, dated 05 / 25 / 2026, page 62 / 728 49 / 335

[00165] Details concerning the most representative cyclic sugar replacement-based vehicles can be found in U.S. Patents Nos. 7,745,608 and 8,017,762, which are incorporated herein by reference in their entirety. Sugar Substitution-Based Monomers (Acyclic)

[00166] Acyclic sugar-substitution based monomers, for example, monomers conjugated to a sugar-substitution base ligand, are also referred to here as ribose substitution monomer subunit monomer (RRMS) compounds. Preferred acyclic carriers may have the formula LCM-3 or LCM-4: LCM-3 W'M·4

[00167] In some embodiments, each of x, yez can be, independently of each other, 0, 1, 2, or 3. In the LCM-3 formula, when yez are different, then the tertiary carbon can have the R or S configuration. In preferred embodiments, x is zero and eyez are each 1 in the LCM-3 formula (e.g., based on serinol), eye z are each 1 in the LCM-3 formula. Each of the LCM-3 or LCM-4 formulas below can be optionally replaced, for example, by hydroxy, alkoxy, per-haloalkyl.

[00168] Details concerning the most representative acyclic sugar-substituted vehicles can be found in U.S. Patents Nos. 7,745,608 and 8,017,762, which are incorporated herein by reference in their entirety.

[00169] In some embodiments, the double-stranded iRNA agent comprises one or more lipophilic moieties conjugated to the 5' end of the sense strand or to the 5' end of the antisense strand. Petition 870260049905, dated 05 / 25 / 2026, p. 63 / 728 50 / 335

[00170] In certain embodiments, the lipophilic portion is conjugated to the 5' end of a strand via a carrier and / or ligand. In one embodiment, the lipophilic portion is conjugated to the 5' end of a strand via a carrier of a formula: Ré um ligando tal como a portion lipophilica.

[00171] In some embodiments, the double-stranded iRNA agent comprises one or more lipophilic moieties conjugated to the 3' end of the sense strand or to the 3' end of the antisense strand.

[00172] In certain embodiments, the lipophilic portion is conjugated to the 3' end of a strand via a carrier and / or ligand. In one embodiment, the lipophilic portion is conjugated to the 3' end of a strand via a carrier of a formula: Petition 870260049905, dated 05 / 25 / 2026, p. 64 / 728 51 / 335 or or . R is a ligand, just like the lipophilic portion.

[00173] In some embodiments, the double-stranded iRNA agent comprises one or more lipophilic moieties conjugated to both ends of the sense strand.

[00174] In some embodiments, the double-stranded iRNA agent comprises one or more lipophilic moieties conjugated to both ends of the antisense strand.

[00175] In some embodiments, the double-stranded iRNA agent comprises one or more lipophilic portions conjugated to the 5' or 3' end of the sense strand and one or more lipophilic portions conjugated to the 5' or 3' end of the antisense strand.

[00176] In some embodiments, the lipophilic portion is conjugated to the terminal end of a ribbon by means of one or more ligands (ties) and / or a vehicle.

[00177] In one embodiment, the lipophilic portion is conjugated to the terminal end of a ribbon by means of one or more ligands (ties).

[00178] In one embodiment, the lipophilic portion is conjugated to the 5' end of the sense or antisense strand via a cyclic vehicle, optionally via one or more intervening ligands (ties). Petition 870260049905, dated 05 / 25 / 2026, p. 65 / 728 52 / 335

[00179] In some embodiments, the lipophilic portion is conjugated to one or more internal positions in at least one strand. Internal positions of a strand refer to the nucleotide at any position on the strand except the terminal position from the 3' end and the 5' end of the strand (e.g., excluding 2 positions: position 1 counting from the 3' end and position 1 counting from the 5' end).

[00180] In one embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, which include all positions except the two terminal positions at each end of the strand (for example, excluding 4 positions: positions 1 and 2 counting from the 3' end and positions 1 and 2 counting from the 5' end). In another embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, which include all positions except the three terminal positions at each end of the strand (for example, excluding 6 positions: positions 1, 2 and 3 counting from the 3' end and positions 1, 2 and 3 counting from the 5' end).

[00181] In one embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, except the region of the cleavage site of the sense strand, for example, the lipophilic portion is not conjugated to positions 9-12 counting from the 5' end of the sense strand, for example, the lipophilic portion is not conjugated to positions 9-11 counting from the 5' end of the sense strand. Alternatively, the internal positions exclude positions 11-13 counting from the 3' end of the sense strand.

[00182] In one embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, which excludes the region of the antisense strand splitting site. For example, the internal positions exclude positions 12-14 counting from the end. Petition 870260049905, dated 05 / 25 / 2026, p. 66 / 728 53 / 335 5' of the nonsensical tape.

[00183] In one embodiment, the lipophilic portion is conjugated to one or more internal positions in at least one strand, excluding positions 11-13 in the sense strand, counting from the 3' end, and positions 12-14 in the antisense strand, counting from the 5' end.

[00184] In one embodiment, one or more lipophilic portions are conjugated to one or more of the following internal positions: positions 4-8 and 13-18 on the sense strip and positions 6-10 and 15-18 on the antisense strip, counting from the 5' end of each strip.

[00185] In one embodiment, one or more lipophilic portions are conjugated to one or more of the following internal positions: positions 5, 6, 7, 15 and 17 on the sense strip and positions 15 and 17 on the antisense strip, counting from the 5' end of each strip.

[00186] In some embodiments, the lipophilic portion is conjugated to a nucleobase, sugar moiety, or internucleoside linkage of the double-stranded iRNA agent. DEFINITIONS

[00187] Unless specific definitions are provided, the nomenclature used in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein is that which is well known and commonly used in the art. Standard techniques for chemical synthesis and chemical analysis may be used. Certain techniques and procedures can be found, for example, in Carbohydrate Modifications in Antisense Research, edited by Sangvi and Cook, American Chemical Society, Washington, DC, 1994; Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 18th edition, 1990; and Antisense Drug Technology, Principles, Strategies, and Applications, edited by Stanley T. Crooke, CRC Press, Boca Raton, Fla.; and Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Har Petition 870260049905, dated 05 / 25 / 2026, page 67 / 728 54 / 335 bor Laboratory Press, 1989, which are hereby incorporated by reference for any purpose. Where permitted, all patents, applications, published applications and other publications and other data referred to throughout the disclosure herein are incorporated by reference in their entirety.

[00188] Unless otherwise indicated, the following terms have the following meanings:

[00189] As used herein, the term target nucleic acid refers to any nucleic acid molecule whose expression or activity is capable of being modulated by an siRNA compound. Target nucleic acids include, but are not limited to, RNA (including, but not limited to, pre-mRNA and mRNA or their portions) transcribed from DNA encoding a target protein, and also cDNA derived from such RNA and miRNA. For example, the target nucleic acid may be a cellular gene (or mRNA transcribed from the gene) whose expression is associated with a particular dysfunction or disease state. In some embodiments, a target nucleic acid may be a nucleic acid molecule of an infectious agent.

[00190] As used herein, the term iRNA refers to an agent that mediates the targeted cleavage of an RNA transcript. These agents associate with a cytoplasmic multiprotein complex known as the RNAi-induced silencing complex (RISC). Agents that are effective in inducing RNA interference are also referred to as siRNA, RNAi agent, or iRNA agent herein. Thus, these terms may be used interchangeably herein. As used herein, the term iRNA includes microRNAs and pre-microRNAs. Furthermore, the compound or compounds of the invention as used herein also refer to the iRNA agent and may be used interchangeably with the iRNA agent.

[00191] The iRNA agent must include a homology region Petition 870260049905, dated 05 / 25 / 2026, pp. 68 / 728 55 / 335 sufficient with the target gene and have sufficient length in terms of nucleotides, such that the iRNA agent, or its fragment, can mediate the downregulation of the target gene. (For ease of exposition, the term nucleotide or ribonucleotide is sometimes used here in reference to one or more monomeric subunits of an iRNA agent. It will be understood here that the use of the term ribonucleotide or nucleotide here may, in the case of a modified RNA or nucleotide substitute, also refer to a modified nucleotide or nucleotide substitute portion at one or more positions.) Thus, the iRNA agent is or includes a region that is at least partially and, in some embodiments, wholly, complementary to the target RNA.Perfect complementarity between the iRNA agent and the target is not necessary, but the match must be sufficient to allow the iRNA agent, or one of its cleavage products, to direct specific sequence silencing, for example, by RNAi cleavage of the target RNA, e.g., mRNA. Complementarity, or the degree of homology with the target strand, is most critical in the antisense strand. Although perfect complementarity, particularly in the antisense strand, is often desired, some embodiments may include, particularly in the antisense strand, one or more, or, for example, 6, 5, 4, 3, 2 or fewer mismatches (with respect to the target RNA). The sense strand only needs to be sufficiently complementary to the antisense strand to maintain the overall double-stranded character of the molecule.

[00192] RNAi agents include: molecules that are long enough to trigger an interferon response (which can be cleaved by Dicer (Bernstein et al. 2001. Nature, 409: 363-366) and enter a RISC (RNAi-induced silencing complex)); and molecules that are short enough that they do not trigger an interferon response (molecules that can also be cleaved by Petition 870260049905, dated 05 / 25 / 2026, pp. 69 / 728 56 / 335 Dicer cleavage (or entry into a RISC), for example, molecules that have a size that allows entry into a RISC, for example, molecules that resemble Dicer cleavage products. Molecules that are short enough that they do not trigger an interferon response are called siRNA agents or shorter iRNA agents herein. siRNA agent or shorter iRNA agent, as used herein, refers to an iRNA agent, for example, a double-stranded RNA agent or single-stranded RNA agent, that is sufficiently short that it does not induce a harmful interferon response in a human cell, for example, has a duplexed region of less than 60, 50, 40, or 30 nucleotide pairs. The siRNA agent, or one of its cleavage products, can downregulate a target gene, for example, by inducing RNAi with respect to a target RNA, where the target may comprise an endogenous or pathogenic target RNA.

[00193] A single-stranded RNA agent, as used herein, is an RNA agent consisting of a single molecule. It may include a duplexed region, formed by intrastrand pairing, for example, it may be, or include, a hairpin or loop structure. Single-stranded RNA agents may be antisense with respect to the target molecule. A single-stranded RNA agent may be long enough that it can enter the RISC and participate in the RISC-mediated cleavage of a target mRNA. A single-stranded RNA agent has at least 14, and in other embodiments, at least 15, 20, 25, 29, 35, 40, or 50 nucleotides in length. In certain embodiments it has less than 200, 100, or 60 nucleotides in length.

[00194] A loop refers to a region of an iRNA strand that is not paired with the opposite nucleotide in the duplex when a section of the iRNA strand forms base pairs with another strand or with or Petition 870260049905, dated 05 / 25 / 2026, pp. 70 / 728 57 / 335 section tra of the same tape.

[00195] The iRNA clamp agents will have a duplex region equal to or at least 17, 18, 19, 29, 21, 22, 23, 24, or 25 nucleotide pairs. The duplex region may be 200, 100, or 50 nucleotides in length. In certain embodiments, the ranges for the duplex region are 15-30, 17-23, 19-23, and 19-21 nucleotide pairs in length. The clamp may have a single-stranded overhang or terminal unpaired region, in some embodiments on the 3' side and, in certain embodiments, antisense of the clamp. In some embodiments, the overhangs are 2-3 nucleotides in length.

[00196] A double-stranded (ds) iRNA agent, as used herein, is an iRNA agent that includes more than one, and in some cases two, strands in which interstrand hybridization can form a duplex region of structure.

[00197] As used herein, the terms siRNA activity and RNAi activity refer to gene silencing by an siRNA.

[00198] As used herein, gene silencing by an RNA interference molecule refers to a decrease in the level of mRNA in a cell for a target gene by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 99% up to and including 100%, and any intermediate integer, from the level of mRNA found in the cell without the presence of the miRNA or RNA interference molecule. In a preferred embodiment, mRNA levels are decreased by at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 99%, up to and including 100% and Petition 870260049905, dated 05 / 25 / 2026, pp. 71 / 728 58 / 335 is any integer between 5% and 100%.

[00199] As used herein, the term modulate gene expression means that the expression of the gene, or the level of the RNA molecule or equivalent RNA molecules encoding one or more proteins or protein subunits, is upregulated or downregulated, such that the expression, level, or activity is greater than or less than that observed in the absence of the modulator. For example, the term modulate can mean inhibit, but the use of the word modulate is not limited to that definition.

[00200] As used herein, gene expression modulation occurs when gene expression, or the level of the RNA molecule or RNA molecules encoding one or more proteins or protein subunits, is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3-fold, 4-fold, 5-fold or more different from that observed in the absence of the siRNA, e.g., RNAi agent. The % and / or magnitude of the difference can be calculated relative to the control or non-control, e.g., [expression with siRNA - expression without siRNA] % difference = -----------------------------------------------------------------------------------------------------------------------------expression without siRNA or [expression with siRNA - expression without siRNA] % difference = -----------------------------------------------------------------------------------------------------------------------------expression without siRNA

[00201] As used herein, the term inhibit, downregulate, or reduce with respect to gene expression means that the expression of the gene, or the level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or the activity of one or more proteins or protein subunits, is reduced below that observed in the absence of a modulator. Gene expression is downregulated when the expression of the gene, or the level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or the activity of Petition 870260049905, dated 05 / 25 / 2026, pp. 72 / 728 59 / 335 one or more proteins or protein subunits is reduced by at least 10% below that of a corresponding unmodulated control and preferably by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99% or, most preferably, 100% (i.e., no gene expression).

[00202] As used herein, the term enhance or overregulate with respect to gene expression means that the expression of the gene, or the level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or the activity of one or more proteins or protein subunits, is increased above that observed in the absence of a modulator. Gene expression is upregulated when the expression of the gene, or the level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or the activity of one or more proteins or protein subunits, is increased by at least 10% relative to a corresponding unmodulated control and preferably by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 100%, 1.1 times, 1.25 times, 1.5 times, 1.75 times, 2 times, 3 times, 4 times, 5 times, 10 times, 50 times, 100 times or more.

[00203] The term increased or augmented as used herein generally means an increase by a statistically significant amount; for avoidance of doubt, increased means an increase of at least 10% compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including an increase of 100% or any increase between 10-100% compared to a reference level, or at least an increase Petition 870260049905, dated 05 / 25 / 2026, pp. 73 / 728 60 / 335 increase of about 2 times, or at least about 3 times, or at least about 4 times, or at least about 5 times, or at least about 10 times, or any increase between 2 times and 10 times or more compared to a reference level.

[00204] The term reduced or reduced as used here generally means a decrease by a statistically significant amount. However, for the avoidance of doubt, reduced means a decrease of at least 10% compared to a reference level, for example a decrease of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a decrease of 100% (i.e., absent level compared to a reference sample), or any decrease between 10-100% compared to a reference level.

[00205] Double-stranded iRNAs comprise two strands of oligonucleotides that are sufficiently complementary to hybridize to form a duplex structure. Generally, the duplex structure is between 15 and 30 base pairs long, more generally between 18 and 25, even more generally between 19 and 24, and most generally between 19 and 21 base pairs long. In some embodiments, longer double-stranded iRNAs of between 25 and 30 base pairs in length are preferred. In some embodiments, shorter double-stranded iRNAs of between 10 and 15 base pairs in length are preferred. In another embodiment, the double-stranded iRNA is at least 21 nucleotides long.

[00206] In some embodiments, the double-stranded iRNA comprises a sense strand and an antisense strand, wherein the antisense strand has a complementarity region that is complementary to at least part of a target sequence and the duplex region has 14-30 nu Petition 870260049905, dated 05 / 25 / 2026, pp. 74 / 728 61 / 335 cleotides in length. Similarly, the region of complementarity with the target sequence is between 14 and 30, more generally between 18 and 25, even more generally between 19 and 24, and most generally between 19 and 21 base pairs in length.

[00207] The phrase antisense ribbon, as used herein, refers to an oligomeric compound that is substantially or 100% complementary to a target sequence of interest. The phrase antisense ribbon includes the antisense region of both oligomeric compounds that are formed from two separate ribbons, as well as unimolecular oligomeric compounds that are capable of forming hairpin or dumbbell-type structures. The terms antisense ribbon and guide ribbon are used interchangeably herein.

[00208] The phrase sense strand refers to an oligomeric compound that has the same nucleoside sequence, in whole or in part, as a target sequence such as a messenger RNA or DNA sequence. The terms sense strand and passenger strand are used interchangeably herein.

[00209] Specifically hybridizable and complementary means that a nucleic acid can form hydrogen bond(s) with another nucleic acid sequence by traditional Watson-Crick or other non-traditional types. With reference to the nucleic acid molecules of the present invention, the binding free energy for a nucleic acid molecule with its complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, for example, RNAi activity. The determination of binding free energies for nucleic acid molecules is well known in the art (see, for example, Turner et al., 1987, CSH Symp. Quant. Biol. Lil pp. 123-133; Frier et al., 1986, Proc. Nat. Acad. Sci. USA 83: 9373-9377; Turner et al., 1987, J. Am. Chem. Soc. 109: 3783-3785). A complementarity percentage indicates the percentage of contiguous residues in Petition 870260049905, dated 05 / 25 / 2026, pp. 75 / 728 62 / 335 a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5, 6, 7, 8, 9, 10 of 10 being 50%, 60%, 70%, 80%, 90%, and 100% complementary). Perfectly complementary or 100% complementarity means that all contiguous residues of one nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence. Less than perfect complementarity refers to the situation in which some, but not all, nucleoside units of two strands can hydrogen bond with each other. Substantial complementarity refers to polynucleotide chains exhibiting 90% or more complementarity, excluding regions of the polynucleotide strands, such as overhangs, that are selected to be non-complementary.Specific binding requires a sufficient degree of complementarity to avoid non-specific binding of the oligomeric compound to non-target sequences under conditions where specific binding is desired, i.e., under physiological conditions in the case of in vivo assays or therapeutic treatment, or, in the case of in vitro assays, under the conditions in which the assays are performed. Non-target sequences typically differ by at least 5 nucleotides.

[00210] In some embodiments, the double-stranded region of a double-stranded iRNA agent is equal to or has at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more nucleotide pairs in length.

[00211] In some embodiments, the antisense strand of a double-stranded iRNA agent is equal to or at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

[00212] In some modes, the sense tape of an agent of Petition 870260049905, dated 05 / 25 / 2026, pp. 76 / 728 63 / 335 double-stranded iRNA is equal to or has at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

[00213] In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each 15 to 30 nucleotides in length.

[00214] In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each 19 to 25 nucleotides in length.

[00215] In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each 21 to 23 nucleotides in length.

[00216] In some embodiments, one strand has at least one 1-5 nucleotide stretch of single-stranded nucleotides in the double-stranded region. A single-stranded nucleotide stretch in the double-stranded region means that at least one nucleotide base pair is present at both ends of the single-stranded stretch. In some embodiments, both strands have at least one 1-5 (e.g., 1, 2, 3, 4, or 5) nucleotide stretch of single-stranded nucleotides in the double-stranded region. When both strands have a stretch of 1-5 (e.g., 1, 2, 3, 4, or 5) single-stranded nucleotides in the double-stranded region, such single-stranded nucleotides may be opposite to each other (e.g., a mismatch stretch) or may be located such that the second strand has no single-stranded nucleotides opposite to the single-stranded iRNAs of the first strand and vice versa (e.g., a single-stranded loop).In some embodiments, single-stranded nucleotides are present within 8 nucleotides from either end, for example 8, 7, 6, 5, 4, 3 or 2 nucleotides from the 5' or 3' end of the complementarity region between the two strands.

[00217] In one embodiment, the double-stranded iRNA agent with. Petition 870260049905, dated 05 / 25 / 2026, pp. 77 / 728 64 / 335 attaches a single-stranded overhang at at least one of its ends. In one embodiment, the single-stranded overhang is 1, 2, or 3 nucleotides in length.

[00218] In one embodiment, the sense strand of the iRNA agent is 21 nucleotides long, and the antisense strand is 23 nucleotides long, wherein the strands form a double-stranded region of 21 consecutive base pairs having a single-stranded overhang 2 nucleotides long at the 3' end.

[00219] In some embodiments, each strand of the double-stranded iRNA has a ZXY structure, as described in PCT Publication No. 2004080406, which is hereby incorporated by reference in its entirety.

[00220] In a certain embodiment, the two strands of the double-stranded oligomeric compound can be linked to each other. The two strands can be linked to each other at both ends or only at one end. By linkage at one end it is understood that the 5' end of the first strand is linked to the 3' end of the second strand or the 3' end of the first strand is linked to the 5' end of the second strand. When the two strands are connected to each other at both ends, the 5' end of the first strand is connected to the 3' end of the second strand and the 3' end of the first strand is connected to the 5' end of the second strand. The two strands can be linked to each other by an oligonucleotide linker including, but not limited to, (N)n; where N is independently a modified or unmodified nucleotide and n is 3-23. In some forms, n is 3-10, for example, 3, 4, 5, 6, 7, 8, 9 or 10.In some embodiments, the oligonucleotide linker is selected from the group consisting of GNRA, (G)4, (U)4, and (dT)4, wherein N is a modified or unmodified nucleotide and R is a modified or unmodified purine nucleotide. Some of the nucleotides in the linker may be involved in interactions. Petition 870260049905, dated 05 / 25 / 2026, pp. 78 / 728 65 / 335 base pairings with other nucleotides in the linker. The two strands can also be linked to each other by a non-nucleoside linker, for example, a linker described herein. It will be appreciated by a person skilled in the art that any modifications or chemical variations of oligonucleotides described herein can be used in the oligonucleotide linker.

[00221] Hairpin and dumbbell-type oligomeric compounds will have a duplex region equal to or having at least 14, 15, 15, 16, 17, 18, 19, 29, 21, 22, 23, 24, or 25 nucleotide pairs. The duplex region may have a length equal to or less than 200, 100, or 50. In some embodiments, the ranges for the duplex region are 1530, 17 to 23, 19 to 23, and 19 to 21 nucleotide pairs in length.

[00222] Hairpin oligomeric compounds may have a single-stranded overhang or terminal unpaired region, in some embodiments on the 3' side and, in some embodiments, antisense of the hairpin. In some embodiments, the protrusions are 1-4, more commonly 2-3, nucleotides in length. Oligomeric hairpin compounds that can induce RNA interference are also referred to as shRNA here.

[00223] In certain embodiments, two oligomeric strands hybridize specifically when there is a sufficient degree of complementarity to avoid nonspecific binding of the antisense compound to non-target nucleic acid sequences under conditions in which specific binding is desired, i.e., under physiological conditions in the case of in vivo assays or therapeutic treatment, and under conditions in which assays are performed in the case of in vitro assays.

[00224] As used herein, stringent hybridization conditions or stringent conditions refer to conditions under which an antisense compound will hybridize with its target sequence, but with a minimum number of other sequences. Stringent conditions are Petition 870260049905, dated 05 / 25 / 2026, pp. 79 / 728 66 / 335 sequence dependent and will be different under different circumstances, and the stringent conditions under which antisense compounds hybridize with a target sequence are determined by the nature and composition of the antisense compounds and the assays in which they are being investigated.

[00225] It is understood in the art that the incorporation of nucleotide affinity modifications may allow for a greater number of mismatches compared with an unmodified compound. Similarly, certain oligonucleotide sequences may be more tolerant of mismatches than other oligonucleotide sequences. A person skilled in the art is able to determine an appropriate number of mismatches between oligonucleotides, or between an oligonucleotide and a target nucleic acid, such as by determining the melting temperature (Tm). Tm or ATm can be calculated by techniques that are familiar to those skilled in the art. For example, the techniques described in Freier et al. (Nucleic Acids Research, 1997, 25, 22: 4429-4443) allow a person skilled in the art to assess nucleotide modifications for their ability to increase the melting temperature of an RNA:DNA duplex. siRNA drawing

[00226] In one embodiment, the iRNA agent of the invention is a 19 nt long double-ended bluntmer, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides at positions 7, 8, 9 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end.

[00227] In one embodiment, the iRNA agent of the invention is a 20 nt long double-ended bluntmer, wherein the sense strand contains at least one motif of three 2'-F modifications. Petition 870260049905, dated 05 / 25 / 2026, page 80 / 728 67 / 335 in three consecutive nucleotides at positions 8, 9, 10 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, 13 from the 5' end.

[00228] In one embodiment, the iRNA agent of the invention is a 21 nt long double-ended bluntmer, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end.

[00229] In one embodiment, the RNA agent of the invention comprises a 21-nucleotide (nt) sense strand and a 23-nucleotide (nt) antisense strand, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5' end; the antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end, wherein one end of the RNA agent is blunt, while the other end comprises a 2 nt overhang. Preferably, the 2 nt overhang is at the 3' end of the antisense. Optionally, the RNA agent further comprises a ligand (e.g., GaINAca).

[00230] In one embodiment, the RNA agent of the invention comprises sense and antisense strands, wherein: the sense strand is 25-30 nucleotide residues in length, wherein, beginning from the 5' terminal nucleotide (position 1), positions 1 to 23 of said first strand comprise at least 8 ribonucleotides; the antisense strand is 36-66 nucleotide residues in length and, beginning from the 3' terminal nucleotide, comprises at least 8 Petition 870260049905, dated 05 / 25 / 2026, page 81 / 728 68 / 335 ribonucleotides in positions paired with positions 1-23 of the sense strand to form a duplex; wherein at least the 3' terminal nucleotide of the antisense strand is not paired with the sense strand, and up to 6 consecutive 3' terminal nucleotides are not paired with the sense strand, thus forming a 3' single-stranded overhang of 1-6 nucleotides; wherein the 5' terminal of the antisense strand comprises 10-30 consecutive nucleotides that are not paired with the sense strand, thus forming a 5' single-stranded overhang of 10-30 nucleotides; whereby at least the 5'-terminal and 3'-terminal nucleotides of the sense strand are base-paired with nucleotides of the antisense strand when the sense and antisense strands are aligned for maximum complementarity, thereby forming a substantially duplexed region between the sense and antisense strands;and the antisense strand is sufficiently complementary to a target RNA along at least 19 ribonucleotides of antisense strand length to reduce target gene expression when said double-stranded nucleic acid is introduced into a mammalian cell; and wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides, wherein at least one of the motifs occurs at or near the cleavage site. The antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at or near the cleavage site.

[00231] In one embodiment, the RNA agent of the invention comprises sense and antisense strands, wherein said RNA agent comprises a first strand having a length of at least 25 and at most 29 nucleotides and a second strand having a length of at most 30 nucleotides with at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end; wherein the Petition 870260049905, dated 05 / 25 / 2026, p. 82 / 728 69 / 335 said 3' end of said first strand and said 5' end of said second strand form a blunt end and said second strand is 1-4 nucleotides longer at its 3' end than the first strand, wherein the duplex region is at least 25 nucleotides in length, and said second strand is sufficiently complementary to a target mRNA along at least 19 nt of said second strand length to reduce target gene expression when said iRNA agent is introduced into a mammalian cell, and wherein dicer cleavage of said iRNA preferentially results in an siRNA comprising said 3' end of said second strand, thereby reducing target gene expression in mammals. Optionally, the iRNA agent further comprises a ligand (e.g., GaINAca).

[00232] In one embodiment, the sense strand of the iRNA agent contains at least one motif of three identical modifications on three consecutive nucleotides, where one of the motifs occurs at the cleavage site on the sense strand. For example, the sense strand may contain at least one motif of three 2'-F modifications on three consecutive nucleotides within 7-15 positions from the 5' end.

[00233] In one embodiment, the antisense strand of the iRNA agent may also contain at least one motif of three identical modifications on three consecutive nucleotides, where one of the motifs occurs at or near the cleavage site on the antisense strand. For example, the antisense strand may contain at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides within 9-15 positions from the 5' end.

[00234] For an iRNA agent having a duplex region 17-23 nt in length, the antisense strand splitting site is typically around positions 10, 11, and 12 from the 5' end. Thus, motifs of three identical modifications can occur. Petition 870260049905, dated 05 / 25 / 2026, page 83 / 728 70 / 335 at positions 9, 10, 11; positions 10, 11, 12; positions 11, 12, 13; positions 12, 13, 14; or positions 13, 14, 15 of the antisense strand, starting the count from the 1st nucleotide from the 5' end of the antisense strand, or starting the count from the 1st paired nucleotide within the duplex region from the 5' end of the antisense strand. The cleavage site on the antisense strand may also change according to the length of the iRNA duplex region from the 5' end.

[00235] In some embodiments, the iRNA agent comprises a sense strand and an antisense strand, each having 14 to 30 nucleotides, wherein the sense strand contains at least two identical three-modification motifs on three consecutive nucleotides, wherein at least one of the motifs occurs at or near the cleavage site within the strand and at least one of the motifs occurs on another portion of the strand that is separated from the motif at the cleavage site by at least one nucleotide. In one embodiment, the antisense strand also contains at least one identical three-modification motif on three consecutive nucleotides, wherein at least one of the motifs occurs at or near the cleavage site within the strand. The motif modification occurring at or near the cleavage site on the sense strand is different from the motif modification occurring at or near the cleavage site on the antisense strand.

[00236] In some embodiments, the iRNA agent comprises a sense strand and an antisense strand, each having 14 to 30 nucleotides, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides, wherein at least one of the motifs occurs at or near the cleavage site on the strand. In one embodiment, the antisense strand also contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at or near the cleavage site. Petition 870260049905, dated 05 / 25 / 2026, page 84 / 728 71 / 335

[00237] In some embodiments, the iRNA agent comprises a sense strand and an antisense strand each having 14 to 30 nucleotides, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5' end, and wherein the antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end.

[00238] In one embodiment, the iRNA agent of the invention comprises mismatch(s) with the target, within the duplex, or combinations thereof. The mismatch may occur in the overhang region or in the duplex region. The base pair may be classified based on its propensity to promote dissociation or fusion (e.g., in the free energy of association or dissociation of a particular pairing, the simplest approach is to examine the pairs on an individual pair basis, although nearest neighbor analysis or similar may also be used). In terms of dissociation promotion: A:U is preferred over G:C; G:U is preferred over G:C; el:C is preferred over G:C (l=inosine).Incompatibilities, for example, non-canonical or non-canonical pairings (as described elsewhere here) are preferred over canonical pairings (A:T, A:U, G:C); and pairings that include a universal basis are preferred over canonical pairings.

[00239] In one embodiment, the iRNA agent of the invention comprises at least one of the first 1, 2, 3, 4, or 5 base pairs within the duplex regions from the 5' end of the antisense strand, which may be independently selected from the group of: A:U, G:U, I:C, and mismatched pairs, for example, non-canonical or non-canonical pairings or pairings that include Petition 870260049905, dated 05 / 25 / 2026, page 85 / 728 72 / 335 a universal base, to promote the dissociation of the antisense tape at the 5' end of the duplex.

[00240] In one embodiment, the nucleotide at position 1 within the duplex region from the 5' end of the antisense strand is selected from the group consisting of A, dA, dU, U, and dT. Alternatively, at least one of the first 1, 2, or 3 base pairs within the duplex region from the 5' end of the antisense strand is an AU base pair. For example, the first base pair within the duplex region from the 5' end of the antisense strand is an AU base pair.

[00241] In one aspect, the invention relates to a double-stranded RNA (dsRNA) agent for inhibiting the expression of a target gene. The dsRNA agent comprises a sense strand and an antisense strand, each strand having 14 to 40 nucleotides. The dsRNA agent is represented by formula (I):

[00242] In formula (I), B1, B2, B3, BΓ, B2', B3' and B4' are each independently a nucleotide containing a selected group modification consisting of 2'-O-alkyl, 2'-substituted alkoxy, 2'-substituted alkyl, 2'-halo, ENA and BNA / LNA. In one embodiment, B1, B2, B3, BΓ, B2', B3' and B4' each contain 2'-OMe modifications. In one embodiment, B1, B2, B3, BΓ, B2', B3' and B4' each contain 2'-OMe or 2'-F modifications. In one embodiment, at least one of B1, B2, B3, BΓ, B2', B3' and B4' contains a 2-O-N-methylacetamide (2-O-NMA) modification.

[00243] C1 is a thermally destabilizing nucleotide placed at a location opposite the seed region of the antisense strand (this Petition 870260049905, dated 05 / 25 / 2026, p. 86 / 728 73 / 335 is located at positions 2-8 from the 5' end of the antisense strand. For example, C1 is at the position on the sense strand that pairs with a nucleotide at positions 2-8 from the 5' end of the antisense strand. In one example, C1 is at position 15 from the 5' end of the sense strand. The C1 nucleotide carries a thermally destabilizing modification that may include an abasic modification; mismatch with the opposite nucleotide in the duplex; and a sugar modification such as a 2'-deoxy modification or an acyclic nucleotide, for example, unblocked nucleic acids (UNA) or glycerol nucleic acid (GNA). In one embodiment, C1 has a thermally destabilizing modification selected from the group consisting of: i) mismatch with the opposite nucleotide on the antisense strand; ii) a selected abasic modification from the group consisting of: ; and iii) selected sugar modification from the group consisting of: In the fixed group, R1 and R2 are independently H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar. In one embodiment, the thermally destabilizing modification at C1 is a selected incompatibility of the group consisting of G:G. Petition 870260049905, dated 05 / 25 / 2026, page 87 / 728 74 / 335 G:A, G:U, G:T, A:A, A:C, C:C, C:U, C:T, U:U, T:T and U:T; and, optionally, at least one nucleobase in the mismatch pair is a 2'-deoxy nucleobase. In one example, the terminally destabilizing modification at C1 is GNA or i

[00244] T1, T1', T2' and T3' each independently represent a nucleotide comprising a modification providing the nucleotide with a steric volume that is less than or equal to the steric volume of a 2'-OMe modification. A steric volume refers to the sum of the steric effects of a modification. Methods for determining the steric effects of a nucleotide modification are known to a person skilled in the art.The modification can be at the 2' position of a ribose sugar in the nucleotide, or a modification in a nucleotide other than ribose, an acyclic nucleotide, or in the nucleotide backbone that is similar or equivalent to the 2' position of the ribose sugar, and provides the nucleotide with a steric volume that is less than or equal to the steric volume of a 2'OMe modification. For example, T1, T1', T2', and T3' are each independently selected from DNA, RNA, LNA, 2'-F, and 2'-F-5'-methyl. In one embodiment, T1 is DNA. In one embodiment, T1' is DNA, RNA, or LNA. In one embodiment, T2' is DNA or RNA. In one embodiment, T3' is DNA or RNA.

[00245] n1, n3 and q1 independently have 4 to 15 nucleotides in length.

[00246] n5, q3 and q7 independently have 1-6 nucleotide(s) in length.

[00247] n4, q2 and q6 independently have 1-3 nucleotide(s) in length; alternatively, n4 is 0.

[00248] q5 independently has 0-10 nucleotide(s) in compress Petition 870260049905, dated 05 / 25 / 2026, page 88 / 728 75 / 335 ment.

[00249] n2e q4 independently have 0-3 nucleotide(s) in length.

[00250] Alternatively, n4 has 0-3 nucleotide(s) in length.

[00251] In one embodiment, n4 can be 0. In one instance, n4 is 0, and q2 and q6 are 1. In another instance, n4 is 0, and q2 and q6 are 1, with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00252] In one modality, n4, q2 and q6 are each 1.

[00253] In one embodiment, n2, n4, q2, q4 and q6 are each 1.

[00254] In one embodiment, C1 is at position 14-17 from the 5' end of the sense strand, when the sense strand is 19-22 nucleotides in length, en4é1. In another embodiment, C1 is at position 15 from the 5' end of the sense strand.

[00255] In one embodiment, T3' starts at position 2 from the 5' end of the antisense tape. In one example, T3' is at position 2 from the 5' end of the antisense tape and q6 is equal to 1.

[00256] In one embodiment, T1' starts at position 14 from the 5' end of the antisense tape. In one example, T1' is at position 14 from the 5' end of the antisense tape and q2 is equal to 1.

[00257] In an exemplary embodiment, T3' starts from position 2 from the 5' end of the antisense tape and TT starts from position 14 from the 5' end of the antisense tape. In an example, T3' starts from position 2 from the 5' end of the antisense tape and q6 equals 1 and T1' starts from position Petition 870260049905, dated 05 / 25 / 2026, page 89 / 728 76 / 335 from the 5' end of the antisense tape and q2 equals 1.

[00258] In one embodiment, TΓ and T3' are separated by 11 nucleotides in length (i.e., not counting the nucleotides T1' and T3').

[00259] In one embodiment, T1' is at position 14 from the 5' end of the antisense strand. In one instance, T1' is at position 14 from the 5' end of the antisense strand and q2 is equal to 1, and the modification at position or positions 2' in a non-ribose, acyclic or backbone provides less steric bulk than a 2'-OMe ribose.

[00260] In one embodiment, T3' is at position 2 from the 5' end of the antisense strand. In one instance, T3' is at position 2 from the 5' end of the antisense strand and q6 is equal to 1, and the modification at position or positions 2' in a non-ribose, acyclic or backbone that provide less than or equal to steric bulk than a 2'-OMe ribose.

[00261] In one embodiment, T1 is at the cleavage site of the sense strand. In one example, T1 is at position 11 from the 5' end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n2 is 1. In an exemplary embodiment, T1 is at the cleavage site of the sense strand at position 11 from the 5' end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n2 is 1.

[00262] In one embodiment, T2' starts at position 6 from the 5' end of the antisense tape. In one example, T2' is at positions 6-10 from the 5' end of the antisense tape, and q4 is 1.

[00263] In an exemplary embodiment, T1 is at the cleavage site of the sense strand, for example, at position 11 from the 5' end of the sense strand, when the sense strand is 19-22 nucleotides in length, en2e1;T1' is at position 14 from the ex Petition 870260049905, dated 05 / 25 / 2026, page 90 / 728 77 / 335 end 5' of the antisense strand, and q2 equals 1, and the TΓ modification is at position 2' of the ribose sugar or at positions on a non-ribose, acidic or backbone that provide less steric bulk than a 2'-OMe ribose; T2' is at positions 6-10 from the 5' end of the antisense strand, and q4 equals 1; and T3' is at position 2 from the 5' end of the antisense strand, and q6 equals 1, and the T3' modification is at position 2' or at positions on a non-ribose, acidic or backbone that provide less than or equal to steric bulk than a 2'-OMe ribose.

[00264] In one embodiment, T2' starts at position 8 from the 5' end of the antisense tape. In one example, T2' starts at position 8 from the 5' end of the antisense tape, and q4 is 2.

[00265] In one embodiment, T2' starts at position 9 from the 5' end of the antisense tape. In one example, T2' is at position 9 from the 5' end of the antisense tape, and q4 is 1.

[00266] In one embodiment, ΒΓ is 2'-OMe or 2'-F, q1 is 9, ΤΓ is 2'F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00267] In one embodiment, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TT is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two internucleotide linkage modifications Petition 870260049905, dated 05 / 25 / 2026, page 91 / 728 78 / 335 phosphorothioate nucleotides at positions 1 and 2 and two internucleotide phosphorothioate linkage modifications within positions 1823 of the antisense strand (counting from the 5' end of the antisense strand).

[00268] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1.

[00269] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00270] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 6, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 7, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1.

[00271] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 6, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 7, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phospho internucleotide linkage modifications Petition 870260049905, dated 05 / 25 / 2026, page 92 / 728 79 / 335 rotothioate within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00272] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1.

[00273] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00274] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 5, T2' is 2'F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; optionally with at least 2 additional TTs at the 3' end of the antisense tape.

[00275] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe Petition 870260049905, dated 05 / 25 / 2026, page 93 / 728 80 / 335 or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 5, T2' is 2'F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; optionally with at least 2 additional TTs at the 3' end of the antisense string; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00276] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1.

[00277] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end).

[00278] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1.

[00279] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, Petition 870260049905, dated 05 / 25 / 2026, page 94 / 728 81 / 335 n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is O, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00280] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1.

[00281] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00282] The dsRNA agent may comprise a phosphorus-containing group at the 5' end of the sense strand or antisense strand. The phosphorus-containing group at the 5' end may be 5'-phosphate (5'-P), 5'-phosphorothioate (5'-PS), 5'-phosphorodithioate (5-PS2), 5'-vinylphosphonate (5'-VP), 5'-methylphosphonate (MePhos), or 5'-deoxy-5'-C-malonyl Petition 870260049905, dated 05 / 25 / 2026, page 95 / 728 82 / 335,0 ΘO)—ι „ Base Θ V ν 7 ο W (OH 0H). When the phosphorus-containing group at the end 5' is vinylphosphonate at the 5' end (5'-VP), the 5'-VP can be the isomer 5-E-VP (i.e., trans-vinylphosphate, OH°), the isomer 5-Z-VP (is- to is, c / s-vinylphosphate,CH°) or mixtures thereof.

[00283] In one embodiment, the dsRNA agent comprises a phosphorus-containing group at the 5' end of the sense strand. In another embodiment, the dsRNA agent comprises a phosphorus-containing group at the 5' end of the antisense strand.

[00284] In one embodiment, the dsRNA agent comprises a 5'-P. In one embodiment, the dsRNA agent comprises a 5'-P on the antisense strand.

[00285] In one embodiment, the dsRNA agent comprises a 5'-PS. In another embodiment, the dsRNA agent comprises a 5'PS on the antisense strand.

[00286] In one embodiment, the dsRNA agent comprises a 5'-VP. In one embodiment, the dsRNA agent comprises a 5'VP on the antisense strand. In one embodiment, the dsRNA agent comprises a 5-E-VP on the antisense strand. In one embodiment, the dsRNA agent comprises a 5-Z-VP on the antisense strand.

[00287] In one embodiment, the dsRNA agent comprises a 5'PS2. In one embodiment, the dsRNA agent comprises a 5'PS2 on the antisense strand.

[00288] In one embodiment, the dsRNA agent comprises a 5-PS2. In another embodiment, the dsRNA agent comprises a 5' Petition 870260049905, dated 05 / 25 / 2026, page 96 / 728 83 / 335 deoxy-5'-C-malonyl in antisense tape.

[00289] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-PS.

[00290] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-P.

[00291] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-VP. 5'-VP can be 5-E-VP, 5-Z-VP, or a combination thereof.

[00292] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-PS2.

[00293] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, TT is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-deoxy-5'-Cmalonyl.

[00294] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe Petition 870260049905, dated 05 / 25 / 2026, page 97 / 728 84 / 335 or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-P.

[00295] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-PS.

[00296] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). Petition 870260049905, dated 05 / 25 / 2026, page 98 / 728 85 / 335 The dsRNA agent also comprises a 5'-VP. The 5'-VP can be 5-E-VP, 5-Z-VP, or a combination thereof.

[00297] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5-PS2.

[00298] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl.

[00299] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-P.

[00300] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, Petition 870260049905, dated 05 / 25 / 2026, page 99 / 728 86 / 335 n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is O, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is O, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-PS.

[00301] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-VP. 5'-VP can be 5-E-VP, 5-Z-VP, or a combination thereof.

[00302] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5-PS2.

[00303] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1. The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl.

[00304] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end).The dsRNA agent comprises size. Petition 870260049905, dated 05 / 25 / 2026, page 100 / 728 87 / 335 also a 5'-P.

[00305] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-PS.

[00306] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-VP. The 5'-VP can be 5-E-VP, 5-Z-VP or a combination thereof.

[00307] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within the 1-5 position of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications. Petition 870260049905, dated 05 / 25 / 2026, page 101 / 728 88 / 335 at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-PS2.

[00308] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl group.

[00309] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1. The dsRNA agent also comprises a 5'-P.

[00310] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1. The dsRNA agent also comprises a 5'-PS.

[00311] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 9, TT is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1. The dsRNA agent also comprises a 5'-VP. The 5'-VP may Petition 870260049905, dated 05 / 25 / 2026, page 102 / 728 89 / 335 be 5-E-VP, 5-Z-VP or a combination thereof.

[00312] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1. The dsRNA agent also comprises a 5-PS2.

[00313] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1. The dsRNA agent also comprises a 5'-deoxy-5'-Cmalonyl.

[00314] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-P.

[00315] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within the 1-5 position of the sense strand (counting from the 5' end of the sense strand) and two internucleotide linkage modifications Petition 870260049905, dated 05 / 25 / 2026, page 103 / 728 90 / 335 phosphorothioate molecules at positions 1 and 2 and two internucleotide linkage modifications of phosphorothioate molecules within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-PS.

[00316] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-VP. The 5'-VP may be 5-E-VP, 5-Z-VP or a combination thereof.

[00317] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5-PS2.

[00318] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2' Petition 870260049905, dated 05 / 25 / 2026, page 104 / 728 91 / 335 F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl.

[00319] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, q7 is 1.0 dsRNA agent also comprises a 5'-P.

[00320] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, q7 is 1.0 dsRNA agent also comprises a 5'-PS.

[00321] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, q7 is 1.0 dsRNA agent also comprises a 5'-VP. The 5'-VP can be 5'E-VP, 5'Z-VP, or a combination thereof.

[00322] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TT is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1. The dsRNA agent also comprises a 5-PS2.

[00323] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, Petition 870260049905, dated 05 / 25 / 2026, page 105 / 728 92 / 335 n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is O, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is O, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1. The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl.

[00324] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-P.

[00325] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-PS.

[00326] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with Petition 870260049905, dated 05 / 25 / 2026, page 106 / 728 93 / 335 two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-VP. The 5'-VP can be 5'-H-VP, 5'Z-VP or a combination thereof.

[00327] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5-PS2.

[00328] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl. Petition 870260049905, dated 05 / 25 / 2026, page 107 / 728 94 / 335

[00329] In one embodiment, 100%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, or 30% of the dsRNA agent of the invention is modified. For example, when 50% of the dsRNA agent is modified, 50% of all nucleotides present in the dsRNA agent contain a modification as described herein.

[00330] In one embodiment, each of the sense and antisense strands of the dsRNA agent is independently modified with acyclic nucleotides, LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-methyl, 2'-O-allyl, 2'-C-allyl, 2'-deoxy, 2'-fluorine, 2'-ON-methylacetamide (2-O-NMA), 2'-O-dimethylaminoethoxyethyl (2-O-DMAEOE), 2'-O-aminopropyl (2O-AP) or 2'-ara-F.

[00331] In one embodiment, each of the sense and antisense strands of the dsRNA agent contains at least two different modifications.

[00332] In one embodiment, the dsRNA agent of Formula (I) further comprises 3' and / or 5' overhang(s) of 1-10 nucleotides in length. In one example, the dsRNA agent of Formula (I) comprises a 3' overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand. In another example, the dsRNA agent has a 5' overhang at the 5' end of the sense strand.

[00333] In one embodiment, the dsRNA agent of the invention does not contain any 2'-F modification.

[00334] In one embodiment, the sense and / or antisense strand of the dsRNA agent comprises one or more blocks of phosphorothioate or methylphosphonate internucleotide bonds. In one example, the sense strand comprises one block of two phosphorothioate or methylphosphonate internucleotide bonds. In another example, the antisense strand comprises two blocks of two phosphorothioate or methylphosphonate internucleotide bonds. For example, the two blocks of internucleotide bonds Petition 870260049905, dated 05 / 25 / 2026, page 108 / 728 95 / 335 nucleotides of phosphorothioate or methylphosphonate are separated by 1618 internucleotide phosphate bonds.

[00335] In one embodiment, each of the sense and antisense strands of the dsRNA agent has 15-30 nucleotides. In one example, the sense strand has 19-22 nucleotides, and the antisense strand has 19-25 nucleotides. In another example, the sense strand has 21 nucleotides, and the antisense strand has 23 nucleotides.

[00336] In one embodiment, the nucleotide at position 1 from the 5' end of the antisense strand in the duplex is selected from the group consisting of A, dA, dU, U, and dT. In one embodiment, at least one of the first, second, and third base pairs from the 5' end of the antisense strand is an AU base pair.

[00337] In one embodiment, the antisense strand of the dsRNA agent of the invention is 100% complementary to a target RNA in order to hybridize with it and inhibits its expression through RNA interference. In another embodiment, the antisense strand of the dsRNA agent of the invention is at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55%, or at least 50% complementary to a target RNA.

[00338] In one aspect, the invention relates to a dsRNA agent as defined herein capable of inhibiting the expression of a target gene. The dsRNA agent comprises a sense strand and an antisense strand, each strand having from 14 to 40 nucleotides. The sense strand contains at least one thermally destabilizing nucleotide, wherein at least one of said thermally destabilizing nucleotides occurs at or near the site opposite the seed region of the antisense strand (i.e., at position 2-8 of the 5' end of the antisense strand). Each of the embodiments and aspects described in this descriptive report relating to the dsRNA represented by formula (I) may also apply to dsRNA containing the nucleotide Petition 870260049905, dated 05 / 25 / 2026, p. 109 / 728 96 / 335 thermally destabilizing.

[00339] The thermally destabilizing nucleotide can occur, for example, between positions 14-17 of the 5' end of the sense strand when the sense strand is 21 nucleotides long. The antisense strand contains at least two modified nucleic acids that are smaller than a sterically demanding 2'-OMe modification. Preferably, the two modified nucleic acids that are smaller than a sterically demanding 2'-OMe are separated by 11 nucleotides in length. For example, the two modified nucleic acids are at positions 2 and 14 of the 5' end of the antisense strand.

[00340] In one embodiment, the dsRNA agent further comprises at least one ASGPR ligand. For example, the ASGPR ligand is one or more GaINAc derivatives attached via a bivalent or trivalent branched ligand, such as: In one example, the ASGPR ligand is attached to the 3' end of the sense tape.

[00341] For example, the dsRNA agent as defined herein may comprise i) a phosphorus-containing group at the 5' end of the sense strand or antisense strand; ii) with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand); and iii) a ligand, such as an ASGPR ligand (for Petition 870260049905, dated 05 / 25 / 2026, page 110 / 728 97 / 335 example, one or more derivatives of GaINAc) at the 5' end or 3' end of the sense or antisense tape. For example, the ligand may be at the 3' end of the sense tape.

[00342] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-P and a targeting ligand. In one embodiment, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00343] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-PS and a targeting ligand. In one embodiment, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the end of the antisense strand. Petition 870260049905, dated 05 / 25 / 2026, page 111 / 728 98 / 335 3' of the sense tape.

[00344] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-VP (e.g., a 5-E-VP, 5-Z-VP, or a combination thereof) and a targeting ligand. In one embodiment, the 5'-VP is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00345] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5-PS2 and a targeting ligand. In one embodiment, the 5-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand. Petition 870260049905, dated 05 / 25 / 2026, page 112 / 728 99 / 335

[00346] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl and a targeting ligand. In one embodiment, the 5'-deoxy-5'-C-malonyl is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00347] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-P and a targeting ligand. In one embodiment, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00348] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, Petition 870260049905, dated 05 / 25 / 2026, page 113 / 728 100 / 335 B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-PS and a targeting ligand. In one embodiment, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00349] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-VP (e.g., a 5-E-VP, 5-Z-VP, or a combination thereof) and a targeting ligand. In one embodiment, the 5'-VP is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00350] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'-F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BT is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the end). Petition 870260049905, dated 05 / 25 / 2026, page 114 / 728 101 / 335 The dsRNA agent comprises a 5-PS2 and a targeting ligand. In one embodiment, the 5-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand. The dsRNA agent also includes two phosphorothioate internucleotide linker modifications at positions 1 and 2 and two phosphorothioate internucleotide linker modifications within positions 18-23 of the antisense strand (counting from the 5' end).

[00351] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl and a targeting ligand. In one embodiment, the 5'-deoxy-5'-C-malonyl is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00352] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the Petition 870260049905, dated 05 / 25 / 2026, page 115 / 728 102 / 335 antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-P and a targeting ligand. In one embodiment, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00353] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-PS and a targeting ligand. In one embodiment, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00354] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-VP (e.g., a Petition 870260049905, dated 05 / 25 / 2026, page 116 / 728 103 / 335 5-E-VP, 5-Z-VP or a combination thereof) and a direction linker. In one embodiment, the 5'-VP is at the 5' end of the antisense tape, and the direction linker is at the 3' end of the sense tape.

[00355] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5-PS2 and a targeting ligand. In one embodiment, the 5-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00356] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl and a targeting ligand. In one embodiment, the 5'-deoxy-5' Petition 870260049905, dated 05 / 25 / 2026, page 117 / 728 104 / 335 C-malonyl is at the 5' end of the antisense tape, and the direction linker is at the 3' end of the sense tape.

[00357] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-P and a targeting ligand. In one embodiment, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00358] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-PS and a targeting ligand. In one embodiment, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00359] In one modality, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, Petition 870260049905, dated 05 / 25 / 2026, page 118 / 728 105 / 335 n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-VP (e.g., a 5-E-VP, 5-ZVP, or a combination thereof) and a targeting ligand. In one embodiment, the 5'-VP is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00360] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5-PS2 and a targeting ligand. In one embodiment, the 5-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00361] In one embodiment, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, BΓ is 2'-OMe or 2'-F, q1 is 9, TΓ is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, Petition 870260049905, dated 05 / 25 / 2026, page 119 / 728 106 / 335 B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F, and q7 is 1; with two phosphorothioate internucleotide linking modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linking modifications at positions 1 and 2 and two phosphorothioate internucleotide linking modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The dsRNA agent also comprises a 5'-deoxy-5'-C-malonyl and a targeting ligand. In one embodiment, the 5'-deoxy-5'-C-malonyl is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[00362] In a particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; and (iii) 2'-F modifications at positions 1, 3, 5, 7, 9 to 11, 13, 17, 19 and 21, and 2'-OMe modifications at positions 2, 4, 6, 8, 12, 14 to 16, 18 and 20 (counting from the 5' end); and (b) a nonsense chain having: (i) a length of 23 nucleotides; (ii) 2'-F modifications at positions 1, 3, 5, 9, 11 to 13, 15, 17, 19, 21 and 23, and 2'-OMe modifications at positions 2, 4, 6 to 8, 10, 14, 16, 18, 20 and 22 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); Petition 870260049905, dated 05 / 25 / 2026, page 120 / 728 107 / 335 in which the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00363] In another particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-F modifications at positions 1, 3, 5, 7, 9 to 11, 13, 15, 17, 19 and 21, and 2'-OMe modifications at positions 2, 4, 6, 8, 12, 14, 16, 18 and 20 (counting from the 5' end); and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) a nonsense chain having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19 and 21 to 23, and 2'-F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18 and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00364] In another particular embodiment, the dsRNA agents of Petition 870260049905, dated 05 / 25 / 2026, page 121 / 728 108 / 335 of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-OMe modifications at positions 1 to 6, 8, 10 and 12 to 21, 2'-F modifications at positions 7 and 9 and a deoxynucleotide (e.g., dT) at position 11 (counting from the 5' end); and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) a nonsense chain having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 7, 9, 11, 13, 15, 17 and 19 to 23, and 2'-F modifications at positions 2, 4 to 6, 8, 10, 12, 14, 16 and 18 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00365] In another particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; Petition 870260049905, dated 05 / 25 / 2026, page 122 / 728 109 / 335 (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-OMe modifications at positions 1 to 6, 8, 10, 12, 14 and 16 to 21, and 2'-F modifications at positions 7, 9, 11, 13 and 15; and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) a nonsense chain having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 5, 7, 9, 11, 13, 15, 17, 19 and 21 to 23, and 2'-F modifications at positions 2 to 4, 6, 8, 10, 12, 14, 16, 18 and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00366] In another particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-OMe modifications in positions 1 to 9 and 12 to 21, and 2'-F modifications in positions 10 and 11; and Petition 870260049905, dated 05 / 25 / 2026, page 123 / 728 110 / 335 (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) a nonsense chain having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19 and 21 to 23, and 2'-F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18 and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00367] In another particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-F modifications at positions 1, 3, 5, 7, 9 to 11 and 13, and 2'-OMe modifications at positions 2, 4, 6, 8, 12 and 14 to 21; and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) a nonsense chain having: Petition 870260049905, dated 05 / 25 / 2026, page 124 / 728 111 / 335 (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 5 to 7, 9, 11 to 13, 15, 17 to 19 and 21 to 23, and 2'-F modifications at positions 2, 4, 8, 10, 14, 16 and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00368] In another particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-OMe modifications at positions 1, 2, 4, 6, 8, 12, 14, 15, 17 and 19 to 21, and 2'-F modifications at positions 3, 5, 7, 9 to 11, 13, 16 and 18; and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) an antisense string having: (i) a length of 25 nucleotides; (ii) 2'-OMe modifications at positions 1, 4, 6, 7, 9, 11 to 13, 15, 17 and 19 to 23, 2'-F modifications at positions 2, 3, 5, 8, 10, 14, 16 and 18 and deoxynucleotides (e.g., dT) at positions 24 and Petition 870260049905, dated 05 / 25 / 2026, p. 125 / 728 112 / 335 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a four-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00369] In another particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-OMe modifications at positions 1 to 6, 8 and 12 to 21, and 2'-F modifications at positions 7, and 9 to 11; and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) a nonsense chain having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3 to 5, 7, 8, 10 to 13, 15 and 17 to 23, and 2'-F modifications at positions 2, 6, 9, 14 and 16 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); Petition 870260049905, dated 05 / 25 / 2026, page 126 / 728 113 / 335 where the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00370] In another particular embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 21 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-OMe modifications at positions 1 to 6, 8 and 12 to 21, and 2'-F modifications at positions 7, and 9 to 11; and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) an antisense string having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15 and 17 to 23, and 2'-F modifications at positions 2, 6, 8, 9, 14 and 16 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22 and between nucleotide positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00371] In another particular embodiment, the dsRNA agents of the present invention comprise: Petition 870260049905, dated 05 / 25 / 2026, page 127 / 728 114 / 335 (a) a sense chain having: (i) a length of 19 nucleotides; (ii) optionally an ASGPR ligand attached to the 3' end, wherein said ASGPR ligand comprises three GaINAc derivatives attached via a trivalent branched ligand; (iii) 2'-OMe modifications at positions 1 to 4, 6 and 10 to 19, and 2'-F modifications at positions 5, and 7 to 9; and (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 (counting from the 5' end); and (b) a nonsense chain having: a length of 21 nucleotides; (ii) 2'-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15 and 17 to 21, and 2'-F modifications at positions 2, 6, 8, 9, 14 and 16 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 19 and 20 and between nucleotide positions 20 and 21 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00372] In one embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 18-23 nucleotides; (ii) three consecutive 2'-F modifications at positions 715; and (b) an antisense string having: Petition 870260049905, dated 05 / 25 / 2026, page 128 / 728 115 / 335 a length of 18-23 nucleotides; (ii) at least 2'-F modifications anywhere in the strand; and (iii) at least two phosphorothioate internucleotide linkages in the first five nucleotides (counting from the 5' end); wherein the dsRNA agents have one or more lipophilic portions conjugated to one or more positions on at least one strand; and have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand; or a blunt end at both ends of the duplex.

[00373] In one embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: (i) a length of 18-23 nucleotides; (ii) less than four 2'-F modifications; (b) a nonsense chain having: a length of 18-23 nucleotides; (ii) fewer than twelve 2'-F modifications; and (iii) at least two phosphorothioate internucleotide linkages in the first five nucleotides (counting from the 5' end); wherein the dsRNA agents have one or more lipophilic portions conjugated to one or more positions on at least one strand; and have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand; or a blunt end at both ends of the duplex.

[00374] In one embodiment, the dsRNA agents of the present invention comprise: (a) a sense chain having: Petition 870260049905, dated 05 / 25 / 2026, p. 129 / 728 116 / 335 (i) a length of 19-35 nucleotides; (ii) less than four 2'-F modifications; (b) a nonsense chain having: a length of 19-35 nucleotides; (ii) fewer than twelve 2'-F modifications; and (iii) at least two phosphorothioate internucleotide linkages in the first five nucleotides (counting from the 5' end); wherein the duplex region has between 19 and 25 base pairs (preferably 19, 20, 21, or 22); and wherein the dsRNA agents have one or more lipophilic portions conjugated to one or more positions on at least one strand; and have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand; or a blunt end at both ends of the duplex.

[00375] In one embodiment, the dsRNA agents of the present invention comprise a sense strand and antisense strands having a length of 15-30 nucleotides; at least two phosphorothioate internucleotide linkages in the first five nucleotides in the antisense strand (counting from the 5' end); wherein the duplex region has between 19 and 25 base pairs (preferably 19, 20, 21 or 22); wherein the dsRNA agents have one or more lipophilic moieties conjugated to one or more positions in at least one strand; and wherein the dsRNA agents have less than 20%, less than 15% and less than 10% of unnatural nucleotide.

[00376] Examples of unnatural nucleotides include acyclic nucleotides, LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-methyl, 2'-O-allyl, 2'-calyl, 2'-deoxy, 2-fluorine, 2'-ON-methylacetamide (2-O-NMA), 2'-O-dimethylaminoethoxyethyl (2-O-DMAEOE), 2'-O-aminopropyl (2-O-AP) or 2-ara-F and others. Petition 870260049905, dated 05 / 25 / 2026, p. 130 / 728 117 / 335

[00377] In one embodiment, the dsRNA agents of the present invention comprise a sense strand and antisense strands having a length of 15-30 nucleotides; at least two phosphorothioate internucleotide linkages in the first five nucleotides in the antisense strand (counting from the 5' end); wherein the duplex region has between 19 and 25 base pairs (preferably 19, 20, 21 or 22); wherein the dsRNA agents have one or more lipophilic moieties conjugated to one or more positions in at least one strand; and wherein the dsRNA agents have more than 80%, more than 85% and more than 90% of natural nucleotide, such as 2'-OH, 2'-deoxy and 2'-OMe are natural nucleotides.

[00378] In one embodiment, the dsRNA agents of the present invention comprise a sense strand and antisense strands having a length of 15-30 nucleotides; at least two phosphorothioate internucleotide linkages in the first five nucleotides in the antisense strand (counting from the 5' end); wherein the duplex region has between 19 and 25 base pairs (preferably 19, 20, 21 or 22); wherein the dsRNA agents have one or more lipophilic moieties conjugated to one or more positions in at least one strand; and wherein the dsRNA agents have 100% natural nucleotides, such as 2'-OH, 2'-deoxy and 2'-OMe are natural nucleotides.

[00379] Examples of lipophilic moieties include, but are not limited to, lipid (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), cholesterol, retinoic acid, cholic acid, adamantanoacetic acid, 1-pyrenebutyric acid, dihydrotestosterone, 1,3-bisO(hexadecyl)glycerol, geranyloxyhexanol, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrimethylammonium chloride, and methylammonium chloride. Petition 870260049905, dated 05 / 25 / 2026, page 131 / 728 118 / 335 tila or phenoxazine.

[00380] In some embodiments, the lipophilic portion is a C6-C30 acid (e.g., hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, octadecanoic acid, oleic acid, linoleic acid, arachidonic acid, cis4,7,10,13,16,19-docosahexanoic acid, vitamin A, vitamin E, cholesterol, etc.) or a C6-C30 alcohol (e.g., hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol, tridecanol, tetradecanol, pentadecanol, hexadecanol, heptadecanol, octadecanol, oleyl alcohol, alcohol linoleic acid, arachidonic alcohol, c / s-4,7,10,13,16,19-docosahexaenolic acid, retinol, vitamin E, cholesterol, etc.).

[00381] In one example, the lipophilic portion is a saturated or unsaturated C6-C18 hydrocarbon chain.

[00382] In one example, the lipophilic portion is docosahexaenoic acid.

[00383] In one embodiment, the dsRNA agents of the present invention have one sense strand and one antisense strand, each strand having 14 to 30 nucleotides, wherein the sequence of the sense strand is represented by formula (I): 5' np-Na-(XXX)i-Nb-Y Y Y-Nb-(ZZZ)j-Na-nq3' (I) where: iej are each independently 0 or 1; each of the peq values ​​is independently 0-6; Each Na independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differentially modified nucleotides; each Nb independently represents a sequence of Petition 870260049905, dated 05 / 25 / 2026, page 132 / 728 119 / 335 oligonucleotides comprising 1, 2, 3, 4, 5 or 6 modified nucleotides; each npe nq independently represents a nucleotide of the protrusion; where Nb and Y do not have the same modification; where XXX, YYY, and ZZZ each independently represent a motif of three identical modifications in three consecutive nucleotides. wherein the dsRNA agents have one or more lipophilic portions conjugated to one or more positions on at least one strand; and wherein the antisense dsRNA strand comprises two blocks of one, two, or three phosphorothioate internucleotide bonds separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 phosphate internucleotide bonds.

[00384] Several publications have described multimeric siRNA, and all can be used with the iRNA of the invention. Such publications include WO2007 / 091269, U.S. Patent No. 7858769, WO2010 / 141511, WO2007 / 117686, WO2009 / 014887 and WO2011 / 031520, which are thus incorporated by reference in their entirety.

[00385] In some embodiments, 100%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35% or 30% of the iRNA agent of the invention is modified.

[00386] In some embodiments, each of the sense and antisense strands of the iRNA agent is independently modified with acyclic nucleotides, LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-methyl, 2'O-allyl, 2'-C-allyl, 2'-deoxy, 2'-fluorine, 2'-ON-methylacetamido (2'-ONMA), 2'-O-dimethylaminoethoxyethyl (2-O-DMAEOE), 2'-O-aminopropyl (2-O-AP) or 2-ara-F. Petition 870260049905, dated 05 / 25 / 2026, p. 133 / 728 120 / 335

[00387] In some embodiments, each of the sense and antisense strands of the iRNA agent contains at least two different modifications.

[00388] In some embodiments, the double-stranded iRNA agent of the invention does not contain any 2'-F modification.

[00389] In some embodiments, the double-stranded iRNA agent of the invention contains one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve 2'-F modification(s). In one example, the double-stranded iRNA agent of the invention contains nine or ten 2'-F modification(s).

[00390] The RNA agent of the invention may additionally comprise at least one phosphorothioate or methylphosphonate internucleotide linkage. The phosphorothioate or methylphosphonate internucleotide linkage modification may occur at any nucleotide of the sense strand or antisense strand or both at any position on the strand. For example, the internucleotide linkage modification may occur at all nucleotides of the sense strand or antisense strand; each internucleotide linkage modification may occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand may contain both internucleotide linkage modifications in an alternating pattern.The alternating pattern of internucleotide linkage modification in the sense strand may be the same as or different from that of the antisense strand, and the alternating pattern of internucleotide linkage modification in the sense strand may deviate from the alternating pattern of internucleotide linkage modification in the antisense strand.

[00391] In one embodiment, the iRNA comprises a phosphorothioate or methylphosphonate internucleotide linkage modification in the overhang region. For example, the overhang region may contain two nucleotides having a phosphorothioate or methylphosphonate internucleotide linkage between the two nucleotides. The linkage modifications in Petition 870260049905, dated 05 / 25 / 2026, page 134 / 728 121 / 335 internucleotides can also be made to link the nucleotides of the overhang with the terminal paired nucleotides within the duplex region. For example, at least 2, 3, 4 or all of the nucleotides of the overhang may be linked via phosphorothioate or methylphosphonate internucleotide linkages, and optionally, there may be additional phosphorothioate or methylphosphonate internucleotide linkages linking the overhang nucleotide with a paired nucleotide that is close to the overhang nucleotide. For example, there may be at least two phosphorothioate internucleotide linkages between the three terminal nucleotides, in which two of the three nucleotides are overhang nucleotides, and the third is a paired nucleotide close to the overhang nucleotide. Preferably, these three terminal nucleotides can be located at the 3' end of the antisense strand.

[00392] In some embodiments, the sense and / or antisense strand of the iRNA agent comprises one or more blocks of phosphorothioate or methylphosphonate internucleotide bonds. In one example, the sense strand comprises one block of two phosphorothioate or methylphosphonate internucleotide bonds. In another example, the antisense strand comprises two blocks of two phosphorothioate or methylphosphonate internucleotide bonds. For example, the two blocks of phosphorothioate or methylphosphonate internucleotide bonds are separated by 1618 phosphate internucleotide bonds.

[00393] In some embodiments, the antisense strand of the iRNA agent of the invention is 100% complementary to a target RNA in order to hybridize with it and inhibits its expression through RNA interference. In another embodiment, the antisense strand of the iRNA agent of the invention is at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55%, or at least 50% complementary to a Petition 870260049905, dated 05 / 25 / 2026, p. 135 / 728 122 / 335 Target RNA.

[00394] In one aspect, the invention relates to an RNA agent capable of inhibiting the expression of a target gene. The RNA agent comprises a sense strand and an antisense strand, each strand having 14 to 40 nucleotides. The sense strand contains at least one thermally destabilizing nucleotide, wherein at least one said thermally destabilizing nucleotide occurs at or near the site opposite the seed region of the antisense strand (i.e., at positions 2-8 of the 5' end of the antisense strand). For example, the thermally destabilizing nucleotide occurs between positions 14-17 of the 5' end of the sense strand when the sense strand is 21 nucleotides long. The antisense strand contains at least two modified nucleic acids that are smaller than a sterically demanding 2'-OMe modification.Preferably, the two modified nucleic acids that are smaller than a sterically demanding 2'-OMe are separated by 11 nucleotides in length. For example, the two modified nucleic acids are at positions 2 and 14 of the 5' end of the antisense strand.

[00395] In some embodiments, the compound of the invention described herein is a miRNA mimetic. In one design, miRNA mimetics are double-stranded molecules (e.g., with a duplex region of about 16 and about 31 nucleotides in length) and contain one or more sequences that have identity with the mature strand of a given miRNA. Double-stranded miRNA mimetics have designs similar to those described above for double-stranded iRNAs. In some embodiments, a miRNA mimetic comprises a duplex region of between 16 and 31 nucleotides and one or more of the following chemical modification patterns: the sense strand contains 2'-O-methyl modifications of nucleotides 1 and 2 (counting from the 5' end of the sense oligonucleotide) and all Cs and Us; the modifications Petition 870260049905, dated 05 / 25 / 2026, p. 136 / 728 123 / 335 of the antisense strand may comprise 2'F modification of all Cs and Us, phosphorylation of the 5' end of the oligonucleotide, and stabilized internucleotide bonds associated with a 3' 2-nucleotide overhang.

[00396] In some embodiments, the compound of the invention described herein is an antimir. In some embodiments, the compound of the invention comprises at least two antimirs covalently linked to each other through a nucleotide-based or non-nucleotide-based linker, for example a linker described in the disclosure, or not covalently linked to each other. The terms antimir, microRNA inhibitor or miR inhibitor are synonymous and refer to oligonucleotides or modified oligonucleotides that interfere with the activity of specific miRNAs.Inhibitors can adopt a variety of configurations including single-stranded, double-stranded (RNA / RNA or RNA / DNA duplex) and hairpin designs. In general, microRNA inhibitors comprise one or more sequences or sequence portions that are complementary or partially complementary to the mature strand(s) of the miRNA to be targeted. Additionally, the miRNA inhibitor may also comprise additional sequences located 5' and 3' relative to the sequence that is the reverse complement of the mature miRNA. The additional sequences may be the reverse complements of the sequences that are adjacent to the mature miRNA in the pri-miRNA from which the mature miRNA is derived, or the additional sequences may be arbitrary sequences (having a mixture of A, G, C, U, or dT). In some embodiments, one or both of the additional sequences are arbitrary sequences capable of forming hairpins.Thus, in some forms, the sequence that is the reverse complement of the miRNA is flanked on the 5' and 3' sides by hairpin structures. MicroRNA inhibitors, when double-stranded, may include mismatches between nucleotides in the strands. Petition 870260049905, dated 05 / 25 / 2026, page 137 / 728 124 / 335 opposite. Furthermore, microRNA inhibitors can be linked to portions of the conjugate in order to facilitate uptake of the inhibitor into a cell.

[00397] MicroRNA inhibitors, including hairpin miRNA inhibitors, are described in detail in Vermeulen et al., DoubleStranded Regions Are Essential Design Components Of Potent Inhibitors of RISC Function, RNA 13: 723-730 (2007) and in WO2007 / 095387 and WO 2008 / 036825, each of which is incorporated herein by reference in its entirety. A person skilled in the art may select a sequence from the database for a desired miRNA and design a useful inhibitor for the methods described herein.

[00398] In some embodiments, the compound of the invention described herein is an antagomir. In some embodiments, the compound of the invention comprises at least two antagomirs covalently linked to each other through a nucleotide-based or non-nucleotide-based linker, for example a linker described in the disclosure, or not covalently linked to each other. Antagomirs are RNA-type oligonucleotides that harbor various modifications for RNase protection and pharmacological properties, such as enhanced tissue and cellular uptake. They differ from normal RNA by, for example, complete 2'-O-methylation of a sugar, a phosphorothioate intersugar linkage, and, for example, a cholesterol moiety at the 3' end.In a preferred embodiment, the antagomir comprises a 2'-O-methyl modification on all nucleotides, a cholesterol moiety at the 3' end, two phosphorothioate intersugar linkages at the first two positions at the 5' end, and four phosphorothioate linkages at the 3' end of the molecule. Antagomirs can be used to efficiently silence endogenous miRNAs by forming duplexes comprising the antagomir and the endogenous miRNA, thereby avoiding miRNA-induced gene silencing. Petition 870260049905, dated 05 / 25 / 2026, page 138 / 728 125 / 335 An example of antagomir-mediated miRNA silencing is the silencing of miR-122, described in Krutzfeldt et al., Nature, 2005, 438: 685-689, which is expressly incorporated by reference herein in its entirety.

[00399] Recent studies have found that dsRNA can also activate gene expression, a mechanism that has been termed small RNA-induced gene activation or RNAa (RNA activation). See for example Li, LC et al. Proc Natl Acad Sci USA. (2006), 103 (46): 17337-42 and Li LC (2008). Small RNA-Mediated Gene Activation. RNA and the Regulation of Gene Expression: A Hidden Layer of Complexity. Caister Academic Press. ISBN 978-1-904455-25-7. dsRNAs targeting gene promoters have been shown to induce potent transcriptional activation of associated genes. Endogenous miRNA causing RNAa has also been found in humans. See E. Nature (2007). 448 (7156): 855-858.

[00400] Another surprising observation is that gene activation by RNAa is long-lasting. The induction of expression has been seen to last for more than ten days. The prolonged effect of RNAa could be attributed to epigenetic changes at dsRNA target sites. In some modes, the RNA activator can increase the expression of a gene. In some modes, the increased gene expression inhibits viability, growth development and / or reproduction.

[00401] Accordingly, in some embodiments, the compound of the invention described herein is activating RNA. In some embodiments, the compound of the invention comprises at least two activating RNAs covalently linked to each other through a nucleotide-based or non-nucleotide-based linker, for example a linker described in the disclosure, or not covalently linked to each other.

[00402] Accordingly, in some forms, the compound of Petition 870260049905, dated 05 / 25 / 2026, p. 139 / 728 The invention described herein is a triplex-forming oligonucleotide (TFO). In some embodiments, the compound of the invention comprises at least two TFOs covalently linked to each other through a nucleotide-based or non-nucleotide-based linker, for example a linker described in the disclosure, or not covalently linked to each other. Recent studies have shown that triplex-forming oligonucleotides can be designed that can recognize and bind to polypurine / polypyrimidine regions in double-stranded helical DNA in a sequence-specific manner. These recognition rules are outlined by Maher III, LJ, et al., Science (1989) vol. 245, pp 725-730; Moser, H. E., et al., Science (1987) vol. 238, pp 645-630; Beal, PA, et al., Science (1992) vol. 251, pp 1360-1363; Conney, M., et al., Science (1988) vol. 241, pp 456-459 and Hogan, ME, et al., Publication EP 375408.Modification of oligonucleotides, such as the introduction of intercalators and intersugar linkage substitutions and optimization of binding conditions (pH and cation concentration), has helped to overcome obstacles inherent in TFO activity such as charge repulsion and instability, and it has recently been shown that synthetic oligonucleotides can be targeted to specific sequences (for a recent review see Seidman and Glazer, J Clin Invest 2003; I 12: 487-94). In general, the triplex-forming oligonucleotide has sequence matching: oligo 3'-AGGT duplex 5'-A GOT duplex 3'-TCGA

[00403] However, it has been shown that A-AT and G-GC triplets have the highest triple helical stability (Reither and Jeltsch, BMC Biochem, 2002, Septl 2, Epub). The same authors demonstrated that TFOs designed according to the A-AT and G-GC rule do not form non-specific triplexes, indicating that triplex formation is in fact es Petition 870260049905, dated 05 / 25 / 2026, page 140 / 728 127 / 335 specific to the sequence.

[00404] Thus, for any given sequence, a triplex-forming sequence can be conceived. Triplex-forming oligonucleotides preferably have at least 15, more preferably 25, even more preferably 30 or more nucleotides in length, up to 50 or 100 nucleotides.

[00405] The formation of the triple helical structure with the target DNA induces steric and functional changes, blocking the initiation and elongation of transcription, allowing the introduction of desired sequence changes in endogenous DNA and resulting in specific downregulation of gene expression. Examples of such suppression of gene expression in cells treated with TFOs include silencing of episomal supFGI and endogenous HPRT genes in mammalian cells (Vasquez et al., Nucl Acids Res. 1999; 27: 1176-81 and Puri, et al., J Biol Chem, 2001; 276: 28991-98) and the sequence-specific and target-specific downregulation of the expression of the transcription factor Ets2, important in the etiology of prostate cancer (Carbone, et al., Nucl Acid Res. 2003; 31: 833-43) and the pro-inflammatory gene ICAM-I (Besch et al., J Biol Chem, 2002; 277: 32473-79).Additionally, Vuyisich and Beal recently showed that sequence-specific TFOs can bind to dsRNA, inhibiting the activity of dsRNA-dependent enzymes such as RNA-dependent kinases (Vuyisich and Beal, Nuc. Acids Res 2000; 28: 2369-74).

[00406] Additionally, TFOs designed according to the principles mentioned above can induce directed mutagenesis capable of effecting DNA repair, thus providing both downregulation and upregulation of endogenous gene expression (Seidman and Glazer, J Clin Invest 2003; 112: 487-94). A detailed description of the design, synthesis, and administration of effective TFOs can be found in U.S. Patent Application Nos. 2003 017068 and Petition 870260049905, dated 05 / 25 / 2026, p. 141 / 728 128 / 335 2003 0096980 in the name of Froehler et al. and 2002 0128218 and 2002 0123476 in the name of Emanuele et al. and U.S. Pat. No. 5,721,138 in the name of Lawn, the contents of which are incorporated herein in their entirety. Nucleic acid modifications

[00407] In some embodiments, the double-stranded RNA agent of the invention comprises at least one nucleic acid modification described herein. For example, at least one modification selected from the group consisting of modified internucleoside linkage, modified nucleobase, modified sugar, and any combinations thereof. Without limitation, such a modification may be present anywhere in the double-stranded RNA agent of the invention. For example, the modification may be present in one of the RNA molecules. Nucleic acid modifications (Nucleobases)

[00408] The naturally occurring base portion of a nucleoside is typically a heterocyclic base. The two most common classes of such heterocyclic bases are purines and pyrimidines. For those nucleosides that include a pentofuranosyl sugar, a phosphate group may be attached to the 2', 3', or 5' hydroxyl portion of the sugar. In the formation of oligonucleotides, these phosphate groups covalently link adjacent nucleosides to each other to form a linear polymeric compound. Within oligonucleotides, the phosphate groups are commonly referred to as forming the internucleoside backbone of the oligonucleotide. The naturally occurring linkage or backbone of RNA and DNA is a 3' to 5' phosphodiester bond.

[00409] In addition to unmodified or natural nucleobases such as the purine nucleobases adenine (A) and guanine (G), and the pyrimidine nucleobases thymine (T), cytosine (C), and uracil (U), many nucleobases or modified nucleobase mimetics are known. Petition 870260049905, dated 05 / 25 / 2026, page 142 / 728 129 / 335 of those described by experts in the art are suitable for the compounds described herein. Unmodified or natural nucleobases can be modified or substituted to provide iRNAs having improved properties. For example, nuclease-resistant oligonucleotides can be prepared with these bases or with synthetic and natural nucleobases (e.g., inosine, xanthine, hypoxanthine, nubularin, isoguanisine, or tubercidine) and any of the oligomer modifications described herein. Alternatively, substituted or modified analogs of any of the above bases and universal bases can be used. When a natural base is substituted for a non-natural and / or universal base, the nucleotide is said to comprise a modified nucleobase and / or a nucleobase modification.Modified nucleobases and / or nucleobase modifications also include natural, non-natural, and universal bases comprising conjugated moieties, for example, a ligand described herein. Preferred conjugate moieties for conjugation with nucleobases include cationic amino groups that can be conjugated to the nucleobase via an alkyl, alkenyl, or an appropriate ligand with an amide linkage.

[00410] An oligomeric compound described herein may also include nucleobase modifications or substitutions (often referred to in the art simply as a base). As used herein, unmodified or natural nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Exemplary modified nucleobases include, but are not limited to, other synthetic and natural nucleobases such as inosine, xanthine, hypoxanthine, nubularin, isoguanisine, tubercidine, 2-(halo)adenine, 2-(alkyl)adenine, 2-(propyl)adenine, 2-(amino)adenine, 2-(aminoalkyl)adenine, 2-(aminopropyl)adenine, 2-(methylthio)-N6-(isopentenyl)adenine, 6-(alkyl)adenine, 6Petition 870260049905, dated 05 / 25 / 2026, page 143 / 728 130 / 335 (methyl)adenine, 7-(deaza)adenine, 8-(alkenyl)adenine, 8-(alkyl)adenine, 8-(alkynyl)adenine, 8-(amino)adenine, 8-(halo)adenine, 8-(hydroxyl)adenine, 8-(thioalkyl)adenine, 8-(thiol)adenine, N6-(isopentyl)adenine, N6-(methyl)adenine, N6, N6(dimethyl)adenine, 2-(alkyl)guanine, 2-(propyl)guanine, 6-(alkyl)guanine, 6-(methyl)guanine, 7-(alkyl)guanine, 7-(methyl)guanine, 7-(deaza)guanine, 8-(alkyl)guanine, 8-(alkenyl)guanine, 8-(alkynyl)guanine, 8-(amino)guanine, 8(halo)guanine, 8-(hydroxyl)guanine, 8-(thioalkyl)guanine, 8(thiol)guanine, N-(methyl)guanine, 2-(thio)cytosine, 3-(deaza)-5(aza)cytosine, 3-(alkyl)cytosine, 3-(methyl)cytosine, 5-(alkyl)cytosine, 5-(alkynyl)cytosine, 5-(halo)cytosine, 5-(methyl)cytosine, 5(propynyl)cytosine, 5-(propynyl)cytosine, 5-(trifluoromethyl)cytosine, 6(azo)cytosine, N4-(acetyl)cytosine, 3-(3-amino-3-carboxypropyl)uracil, 2-(thio)uracil, 5-(methyl)-2-(thio)uracil, 5-(methylaminomethyl)-2(thio)uracil, 4-(thio)uracil,5-(methyl)-4-(thio)uracil, 5-(methylaminomethyl)4-(thio)uracil, 5-(methyl)-2,4-(dithio)uracil, 5-(methylaminomethyl)-2,4(dithio)uracil, 5-(2-aminopropyl)uracil, 5-(alkyla,) 5(alkynyl)uracil, 5-(allylamino)uracil, 5-(aminoallyl)uracil, 5(aminoalkyl)uracil, 5-(guanidinioalkyl)uracil, 5-(1,3-diazol-1alkyl)uracil, 5-(cyanoalkyl)uracil, 5-(alkyl)uracil 5-(dimethylaminoalkyl)uracil, 5-(halo)uracil, 5-(methoxy)uracil, uracil-5-oxyacetic acid, 5-(methoxycarbonylmethyl)-2-(thio)uracil, 5(methoxycarbonyl-methyl)uracil, 5-(propinyl)uracil 5-(propinyl)uracil, 5-(trifluoromethyl)uracil, 6-(azo)uracil, di-hydrouracil, N3(methyl)uracil, 5-uracil (i.e., pseudouracil), 2-(thio)pseudouracil, 4-(thio)pseudouracil, 2,4-(dithio)pseudouracil, 5-(alkyl)pseudouracil, 5-(methyl)pseudouracil, 5-(alkyl)-2-(thio)pseudouracil, 5-(methyl)-2(thio)pseudouracil, 5-(alkyla)-4-pseudouracil, 5-(methyl)-4(thio)pseudouracil, 5-(alkyl)-2,4-(dithio)pseudouracil, 5-(methyl)-2,4-Petition 870260049905, dated 05 / 25 / 2026, page 144 / 728, 131 / 335 (dithio)pseudouracil, 1-substituted pseudouracil, 2(thio)-pseudouracil 1-substituted, 4-(thio)pseudouracil 1-substituted, 2,4(dithio)pseudouracil 1-substituted, l-(aminocarbonylethylenyl)pseudouracil, 1 -(aminocarbonylethylenyl)-2(thio)-pseudouracyl, 1-(aminocarbonylethylenyl)-4-(thio)pseudouracyl, l-(aminocarbonylethylenyl)2,4-(dithio)pseudouracyl, l-(aminoalkylaminocarbonylethylenyl)pseudouracyl, 1-(aminoalkylamino-carbonylethylenyl)-2(thio)pseudouracyl, 1-(aminoalkylaminocarbonylethylenyl)-4(thio)pseudouracil, 1-(aminoalkylaminocarbonylethylene)-2,4-(dithio)pseudouracil, 1,3-(diaza)-2-(oxo)-phenoxazine-1-yl, 1-(aza)-2-(thio)3-(aza)-phenoxazine-1-yl, 1,3-(diaza)-2-(oxo)-fenthiazin-1-yl, 1-(aza)-2-(thio)-3-(aza)-fenthiazin-1-yl, 1,3-(diaza)-2-(oxo)-phenoxazine-1-yl 7-substituted, 1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl 7-substituted, 1,3-(diaza)-2-(oxo)-fenthiazin-1-yl 7-substituted, 1 7-substituted -(aza)-2-(thio)-3-(aza)phenthiazin-1-yl, 7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)phenoxazin-1 -yl, 7-(aminoalkyl-hydroxy)-1-(aza)-2-(thio)-3-(aza)phenoxazin-1 -yl, 7-(aminoalkyl-hydroxy)-1,3-(diaza)-2-(oxo)-phenthiazin-1 -yl, 7-(aminoalkyl-hydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1 -yl, 7(guanidinioalkyl-hydroxy)-l ,3-(diaza)-2-(oxo)-phenoxazin-1-yl, 7(guanidinioalkyl-hydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl, 7(guanidinioalkyl-hydroxy)-l ,3-(diaza)-2-(oxo)-phenthiazin-1-yl, 7(guanidinioalkyl-hydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1-yl, 1,3,5(triaz)-2,6-(dioxa)-naphthalene, inosine, xanthine, hypoxanthine, nubularin, tubercidin, isoguanisine, inosinyl, 2-aza-inosinyl, 7-deazainosinyl, nitroimidazolyl, nitropyrazolyl, nitrobenzimidazolyl, nitroindazolyl, aminoindolyl, pyrrolopyrimidinyl, 3-(methyl)isocarbostyryl, 5(methyl)isocarbostyryl, 3-(methyl)-7-(propynyl)isocarbostyryl, 7(aza)indolyl, 6-(methyl)-7-(aza)indolyl, imidizopyridinyl, 9-(methyl)imidizopyridinyl, pyrrolopyrizinyl, isocarbostyryl,7(propynyl)isocarbostyrylyl, propynyl-7-(aza)indolyl, 2,4,5-(trimethyl)phenyl, Petition 870260049905, dated 05 / 25 / 2026, page 145 / 728 132 / 335 4-(methyl)indolyl, 4,6-(dimethyl)indolyl, phenyl, naphthalenyl, anthracenyl, phenanthracenyl, pyrenyl, stilbenyl, tetracenyl, pentacenyl, difluorotolyl, 4-(fluoro)-6-(methyl)benzimidazole, 4-(methyl)benzimidazole, 6(azo)thymine, 2-pyridinone, 5-nitroindole, 3-nitropyrrole, 6-(aza)pyrimidine, 2(amino)purine, 2,6-(diamino)purine, 5-substituted pyrimidines, N2-substituted purines, purines N6-substituted, O6-substituted purines, 1,2,4-substituted triazois, pyrrolo-pyrimidin-2-on-3-yl, 6-phenyl-pyrrolopyrimidin-2-on-3-yl, para-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl, ortho-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl, bis-ortho-substituted6-phenyl-pyrrolo-pyrimidin-2-on-3-yl, para-(aminoalkyl-hydroxy)-6-phenylpyrrolo-pyrimidin-2-on-3-yl, ortho-(aminoalkyl-hydroxy)-6-phenyl-pyrrolopyrimidin-2-one-3-yl, bis-ortho-(aminoalkyl-hydroxy)-6-phenyl-pyrrolopyrimidin-2-one-3-yl, pyridopyrimidin-3-yl, 2-oxo-7-amino-pyridopyrimidin3-yl,2-oxo-pyridopyrimidine-3-yl or any of its O-alkylated or N-alkylated derivatives. Alternatively, substituted or modified analogs of any of the above bases and universal bases may be used.

[00411] As used herein, a universal nucleobase is any nucleobase that can base-pair with all four naturally occurring nucleobases without substantially affecting fusion behavior, recognition by intracellular enzymes, or iRNA duplex activity. Some illustrative universal nucleobases include, but are not limited to, 2,4-difluorotoluene, nitropyrrole, nitroindolyl, 8-aza-7-deaza-adenine, 4-fluoro-6-methylbenzimidazole, 4-methylbenzimidazole, 3-methylisocarbostyrylyl, 5-methylisocarbostyrylyl, 3-methyl-7-propynyl, isocarbostyrylyl, 7-azaindolyl, 6-methyl-7-azaindolyl, imidizopyridinyl, 9-methylimidizopyridinyl, pyrrolopirizinyl, isocarbostyrylyl, 7-propynylisocarbostyrylyl, propynyl-7-azaindolyl, 2,4,5-trimethylphenyl, 4-methylinolyl, 4,6-dimethylindolyl, phenyl, naphthalenyl, anthracenyl, phenanthracenyl, pyrenyl, Petition 870260049905, dated 05 / 25 / 2026, page 146 / 728 133 / 335 stilbenyl, tetracenyl, pentacenyl and their structural derivatives (see, for example, Loakes, 2001, Nucleic Acids Research, 29, 2437-2447).

[00412] Additional nucleobases include those disclosed in U.S. Pat. No. 3,687,808; those disclosed in International Application No. PCT / US09 / 038425, filed March 26, 2009; those disclosed in the Concise Encyclopedia of Polymer Science and Engineering, pages 858-859, Kroschwitz, JI, ed. John Wiley & Sons, 1990; those disclosed by English et al., Angewandte Chemie, International Edition, 1991, 30, 613; those disclosed in Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijin, P. Ed. Wiley-VCH, 2008; and those published by Sanghvi, YS, Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, ST and Lebleu, B., Eds., CRC Press, 1993. The contents of all the above are incorporated herein by reference.

[00413] In certain embodiments, a modified nucleobase is a nucleobase that is quite similar in structure to the parent nucleobase, such as, for example, a 7-desaza purine, a 5-methyl cytosine, or a G-clamp. In certain embodiments, nucleobase mimetics include more complicated structures, such as, for example, a tricyclic phenoxazine nucleobase mimetic. Methods for preparing the modified nucleobases noted above are well known to those skilled in the art. Nucleic acid (sugar) modifications

[00414] The double-stranded RNA agent of the invention provided herein may comprise one or more (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more) monomers, including a nucleoside or nucleotide, having a modified sugar moiety. For example, the furanosyl sugar ring of a nucleoside may be modified in a number of ways including, but not limited to, addition of a substituent group, bridging of two atoms. Petition 870260049905, dated 05 / 25 / 2026, page 147 / 728 134 / 335 non-geminal annulus molecules combine to form a blocked nucleic acid or bicyclic nucleic acid. In certain embodiments, oligomeric compounds comprise one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more) monomers that are LNA.

[00415] In some embodiments of a blocked nucleic acid, the 2' furanosyl position is connected to the 4' position by a ligand selected independently of -[C(R1)(R2)]n-, -[C(R1)(R2)]nO-, [C(R1 )(R2)]nN(R1)-, -[C(R1 )(R2)]nN(R1 )-O-, -[C(R1 R2)]nON(R1)-, -C(R1 )=C(R2)-O-, -C(R1)=N-, -C(R1)=NO-, -C(=NR1)-, -C(=NR1)O-, -C(=O)-, -C(=O)O-, -C(=S)-, -C(=S)O-, -C(=S)S-, -O-, -Si(R1)2-, S(=O)xeN(R1)-; in which: x is 0, 1, or 2; n is 1, 2, 3 or 4; Each R1 and R2 is independently H, a protecting group, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, alkenyl O2O12, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C2O aryl, substituted C5-C2O aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJ1, NJ1J2, SJ1, N3, C0OJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1) or sulfoxyl (S(=O)-J1); and each J1 and J2 is independently H, C1-C12 alkyl, substituted C1-C12 alkyl, O2-O12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted O2-O12 alkynyl, C5-C2O aryl, substituted C5-C12 aryl, acyl (C(=O)-H), substituted acyl, a heterocyclic radical, a substituted heterocyclic radical, O1O12 aminoalkyl, substituted C1-C12 aminoalkyl or a protecting group.

[00416] In some embodiments, each of the ligands of the LNA compounds is independently -[C(R1)(R2)]n-, Petition 870260049905, dated 05 / 25 / 2026, p. 148 / 728 135 / 335 -[C(R1)(R2)]n-O-, -C(R1R2)-N(R1)-O- or -C(R1R2)-ON(R1)-. In another embodiment, each of said ligands is independently 4'CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R1)-2' and 4'-CH2-N(R1)-O-2- wherein each R1 is independently H, a protecting group or C1-C12 alkyl.

[00417] Certain LNAs have been prepared and described in the patent literature as well as in the scientific literature (Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 36073630; Wahlestedt et al., Proc. Natl. Acad. Sci. USA, 2000, 97, 56335638; Kumar et al., Bioorg. Med. Chem. 10035-10039); Examples of issued US patents and published applications disclosing LNAs include, for example, US Pat. Nos. 7,053,207; 6,268,490; 6,770,748; 6,794,499; 7,034,133; and 6,525,191; and U.S. Pre-Grant Publications Nos. 20040171570; 2004-0219565; 2004-0014959; 2003-0207841; 20040143114; and 20030082807.

[00418] Also provided herein are LNAs in which the 2'-hydroxyl group of the ribosyl sugar ring is linked to the 4'-carbon atom of the sugar ring thereby forming a methyleneoxy linkage (4'-CH2-O-2') to form the bicyclic sugar moiety (reviewed in Elayadi et al., Curr. Opinion Invens. Drugs, 2001, 2, 558-561; Braasch et al., Chem. Bio!, 2001, 8 1-7; and Orum et al., Curr. Opinion Mol. Then, 2001, 3, 239-243; see also U.S. Pat. Nos. 6,268,490 and 6,670,461). The linkage can be a methylene group (-CH2-) bridging the oxygen atom 2' and the carbon atom 4', for which the term LNA of methylene-oxy (4'-CH2-O-2') is used for the bicyclic portion; in the case of an ethylene group in this position, the term LNA of ethylene-oxy (4'-CH2CH2-O-2') is used (Singh et al., Chem. Commun., 1998, 4, 455-456; Morita et al., Bioorganic Medicinal Chemistry, 2003). Petition 870260049905, dated 05 / 25 / 2026, page 149 / 728 136 / 335 11, 2211-2226). Methylene-oxy(4'-CH2-O-2') LNA and other bicyclic sugar analogs exhibit very high duplex thermal stabilities with complementary DNA and RNA (Tm = +3 to +10 °C), stability in the direction of 3'-exonucleolytic degradation, and good solubility properties. Potent and non-toxic antisense oligonucleotides comprising BNAs have been described (Wahlestedt et al., Proc. Natl. Acad. Sci. USA, 2000, 97, 5633-5638).

[00419] An isomer of methylene-oxy(4'-CH2-O-2') LNA that has also been discussed is alpha-L-methylene-oxy(4'-CH2-O-2') LNA, which has been shown to have superior stability against a 3' exonuclease. Alpha-L-methylene-oxy(4'-CH2-O-2') LNAs have been incorporated into antisense gappers and chimeras that have shown potent antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372).

[00420] The synthesis and preparation of LNA monomers of methylene-oxy(4'-CH2-O-2') adenine, cytosine, guanine, 5-methyl-cytosine, thymine and uracil, together with their oligomerization and nucleic acid recognition properties have been described (Koshkin et al., Tetrahedron, 1998, 54, 3607-3630). BNAs and their preparation are also described in WO 98 / 39352 and WO 99 / 14226.

[00421] Analogs of methylene-oxy (4'-CH2-O-2'), phosphorothioate-methylene-oxy (4'-CH2-O-2') and 2'-thio-LNA have also been prepared (Kumar et al., Bioorg. Med. Chern. Lett., 1998, 8, 2219-2222). The preparation of blocked nucleoside analogs comprising oligodeoxyribonucleotide duplexes as substrates for nucleic acid polymerases has also been described (Wengel et al., NO 99 / 14226). Furthermore, the synthesis of 2'-amino-LNA, a novel conformationally restricted high-affinity oligonucleotide analog, was described in the technique (Singh et al., J. Org. Chem., 1998, 63, 10035-10039). Additionally, 2'-Amino- and 2'-methylamino-LNAs were prepared and the thermal stability of their duplexes with strands of Petition 870260049905, dated 05 / 25 / 2026, page 150 / 728 137 / 335 Complementary RNA and DNA have been previously reported.

[00422] Modified sugar moieties are well known and can be used to alter, typically increase, the affinity of the antisense compound for its target and / or increase resistance to nucleases. A representative list of preferred modified sugars includes, but is not limited to, bicyclic modified sugars, including methylene-oxy LNA (4-CH2-O-2') and ethylene-oxy ENA (4'-(CH2)2-O-2' bridge); substituted sugars, especially 2'-substituted sugars having a 2'-F, 2-OCH3, or 2'-O(CH2)2OCH3 substituent group; and 4'-thio-modified sugars. Sugars may also be substituted by sugar mimetic groups, among others. Methods for the preparation of modified sugars are well known to those skilled in the art. Some representative patents and publications that teach the preparation of such modified sugars include, but are not limited to, U.S. Pat. Nos.4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; 5,792,747; 5,700,920; 6,531,584; and 6,600,032; and WO 2005 / 121371.

[00423] Examples of oxy-2'-hydroxyl group modifications include alkoxy or aryloxy (OR, e.g., R = H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar); polyethylene glycols (PEG), O(CH2CH2O)nCH2CH2OR, n = 1-50; locked nucleic acids (LNA) in which the furanose portion of the nucleoside includes a bridge connecting two carbon atoms in the furanose ring, thereby forming a bicyclic ring system; O-AMINE or O-(CH2)nAMINE (n = 1-10, AMINE = NH2; alkylamino, dialkylamino, heterocyclyl, arylamino, diarylamino, heteroarylamino, di-heteroarylamino, ethylenediamine, or polyamino); and O-CH2CH2(NCH2CH2NMe2)2. Petition 870260049905, dated 05 / 25 / 2026, page 151 / 728 138 / 335

[00424] Deoxy modifications include hydrogen (i.e., deoxyribose sugars, which are particularly relevant to single-stranded overhangs); halo (e.g., fluorine); amino (e.g., NH2; alkylamino, dialkylamino, heterocyclyl, arylamino, diarylamino, heteroarylamino, diheteroarylamino, or amino acid); NH(CH2CH2NH)nCH2CH2-AMINE (AMINE = NH2; alkylamino, dialkylamino, heterocyclyl, arylamino, diarylamino, heteroarylamino, or diheteroarylamino); -NHC(O)R (R = alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar); cyano; mercapto; alkyl-thioalkyl; thioalkoxy; thioalkyl; alkyl; cycloalkyl; aryl; alkenyl and alkynyl, which may be optionally substituted by, for example, an amino resource.

[00425] Other suitable modifications of 2', for example, modified MOE, are described in U.S. Patent Application Publication No. 20130130378, the contents of which are incorporated herein by reference.

[00426] A modification at the 2' position may be present in the arabinose configuration. The term arabinose configuration refers to the placement of a substituent at the C2' of ribose in the same configuration as the 2'-OH is in arabinose.

[00427] The sugar may comprise two different modifications at the same carbon in the sugar, for example, gem modification. The sugar group may also contain one or more carbons that have a stereochemical configuration opposite to that of the corresponding carbon in ribose. Thus, an oligomeric compound may include one or more monomers containing, for example, arabinose, as the sugar. The monomer may have an alpha linkage at the T position in the sugar, for example, alpha-nucleosides. The monomer may also have the opposite configuration at the 4' position, for example, O5' and H4', or substituents replacing the same are exchanged with each other. When C5' and OH4' or Petition 870260049905, dated 05 / 25 / 2026, page 152 / 728 139 / 335 substituents replacing the same are exchanged among themselves, it is said that the sugar is modified at position 4'.

[00428] The double-stranded RNA agent of the inventions described herein may also include abasic sugars, i.e., a sugar that does not have a nucleobase at C-1' or has other chemical groups in place of a nucleobase at C1'. See, for example, U.S. Pat. No. 5,998,203, the contents of which are incorporated herein in their entirety. These abasic sugars may also additionally contain modifications to one or more of the constituent sugar atoms. The double-stranded RNA agent of the invention may also contain one or more sugars that are the L isomer, for example, L-nucleosides. The modification to the sugar group may also include substitution of the 4'-O by a sulfur, optionally substituted nitrogen, or CH2 group. In some embodiments, the linkage between CT and the nucleobase is in the α configuration.

[00429] Sugar modifications may also include acyclic nucleotides, where a C-C bond between ribose carbons (e.g., C1'-C2', C2'-C3', C3'-C4', C4'-O4', C1'-O4') is absent and / or at least one of the ribose carbons or oxygens (e.g., CT, O2', C3', O4', or O4') is independently or in combination absent from the nucleotide. In some embodiments, the acyclic nucleotide III jx / x / xr jx / x / xn uxruxr where B is a modified or unmodified nucleobase, R1 and R2 are independently H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar.

[00430] In some embodiments, the sugar modifications are selected from the group consisting of 2'-H, 2'-O-Me (2'-O-methyl), 2'-O Petition 870260049905, dated 05 / 25 / 2026, p. 153 / 728 140 / 335 MOE (2'-O-methoxyethyl), 2'-F, 2'-O-[2-(methylannino)-2-oxoethyl] (2-ONMA), 2'-S-methyl, 2'-O-CH2-(4'-C) (LNA), 2'-O-CH2CH2-(4'-C) (ENA), 2'-O-aminopropyl (2-O-AP), 2'-O-dimethylaminoethyl (2-O-DMAOE), 2'O-dimethylaminopropyl (2-O-DMAP), 2'-O-dimethylaminoethyloxyethyl (2'-ODMAEOE) and gem 2'-OMe / 2'F with 2'-O-Me in the arabinose configuration.

[00431] It is to be understood that when a particular nucleotide is linked through its 2' position to the following nucleotide, the sugar modifications described herein may be placed at the 3' position of the sugar for that particular nucleotide, for example, the nucleotide that is linked through its 2' position. A modification at the 3' position may be present in the xylose configuration. The term xylose configuration refers to the placement of a substituent at the C3' of ribose in the same configuration as the 3'-OH is in the xylose sugar.

[00432] The hydrogen attached to C4' and / or C1' may be replaced by an optionally substituted linear or branched alkyl, optionally substituted alkenyl, optionally substituted alkynyl, wherein the alkyl, alkenyl and alkynyl skeleton may contain one or more O, S, S(O), SO2, N(R'), C(O), N(R')C(O)O, OC(O)N(R'), CH(Z'), phosphorus-containing linkage, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic or optionally substituted cycloalkyl, where R' is hydrogen, acyl or optionally substituted aliphatic, Z' is selected from the group x Nx-s N NN\ / N~R21'N N' ~N-R21 consisting of ORn, CORn, CO2Rn, η ,nx'NR21, NR2iRsi, CONR2iR3i, CON(H)NR2i R31, ONR2-iR3i, CON(H)N=CR4iR5i, N(R2i)C(=NR3i)NR2iR3i, N(R2i)C(O)NR2iR3i, N(R2i)C(S)NR2iR3i, OC(O)NR2iR3i, SC(O)NR2iR3i, N(R2i)C(S)ORh, N(R2i)C(O)ORh, N(R2i)C(O)SRh, N(R2i)N=CR4iR5i, ON=CR4iR5i, Petition 870260049905, dated 05 / 25 / 2026, p. 154 / 728 141 / 335 SO2R11, SORn, SR11 and substituted or unsubstituted heterocyclic; R21 and R31 are for each occurrence independently hydrogen, acyl, unsubstituted or substituted aliphatic, aryl, heteroaryl, heterocyclic, OR11, COR11, CO2R11 or NR11Rn'; or R21 and R31, taken together with the atoms to which they are attached, form a heterocyclic ring; R41 and R51 are for each occurrence independently hydrogen, acyl, unsubstituted or substituted aliphatic, aryl, heteroaryl, heterocyclic, ORn, CORn or CO2R11 or NR11R11'; and Rn and Rn' are independently hydrogen, aliphatic, substituted aliphatic, aryl, heteroaryl or heterocyclic. In some embodiments, the hydrogen attached to the C4' of the 5' terminal nucleotide is substituted.

[00433] In some embodiments, C4' and C5' together form an optionally substituted heterocycle, preferably comprising at least one -PX(Y)-, wherein X is H, OH, OM, SH, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkylthio, optionally substituted alkylamino or optionally substituted dialkylamino, where M is independently for each occurrence an alkali metal or transition metal with an overall charge of +1; and Y is O, S or NR', where R' is optionally substituted aliphatic hydrogen. Preferably, this modification is at the 5'-terminus of the iRNA.

[00434] In certain embodiments, LNAs include a bicyclic nucleoside having the formula: in which: Bx is a heterocyclic base moiety; Ti is H or a hydroxyl protecting group; T2 is H, a protecting hydroxyl group or a phosphorus group reacts. Petition 870260049905, dated 05 / 25 / 2026, page 155 / 728 142 / 335 active; Z is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, substituted C1-C6 alkyl, substituted C2-C6 alkenyl, substituted C2-C6 alkynyl, acyl, substituted acyl, or substituted amide.

[00435] In some embodiments, each of the substituted groups is independently mono- or polysubstituted by optionally protected substituent groups independently selected from halogen, oxo, hydroxyl, OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 and CN, wherein each J1, J2 and J3 is independently H or C1-Ce alkyl, and X is O, S or NJ1.

[00436] In certain such embodiments, each of the substituted groups is independently mono- or polysubstituted by independently selected halogen, oxo, hydroxyl, OJ1, NJ1J2, SJ1, N3, OC(=X)J1 and NJ3C(=X)NJ1 J2 substituent groups, wherein each J1, J2 and J3 is independently H, Ci-Ce alkyl or substituted CiCe alkyl and X is O or NJ1.

[00437] In certain embodiments, the Z group is a Ci-Ce alkyl substituted by one or more Xx, wherein each Xx is independently OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 or CN; wherein each J1, J2 and J3 is independently H or Ci-Ce alkyl, and X is O, S or NJ1. In another embodiment, the Z group is a Ci-Ce alkyl substituted by one or more XX, wherein each Xx is independently halo (e.g., fluorine), hydroxyl, alkoxy (e.g., CH3O-), substituted alkoxy or azide.

[00438] In certain embodiments, the Z group is -CH2XX, where Xx is OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 or CN; where each J1, J2 and J3 is independently H or C1Ce alkyl, and X is O, S or NJ1. In another embodiment, the Z group is -CH2XX, where Xx is halo (e.g., fluorine), hydroxyl, alkoxy (e.g., CH3O-) or azide. Petition 870260049905, dated 05 / 25 / 2026, p. 156 / 728 143 / 335

[00439] In certain such modalities, the group Z is in configuration (R):

[00440] In certain such modalities, the Z group is in configuration (S):

[00441] In certain embodiments, each Ti and T2 is a hydroxyl protecting group. A preferred list of hydroxyl protecting groups includes benzyl, benzoyl, 2,6-dichlorobenzyl, l-butyldimethylsilyl, l-butyldiphenylsilyl, mesylate, tosylate, dimethoxytrityl (DMT), 9-phenylxanthine-9-yl (Pixyl) and 9(p-methoxyphenyl)xanthine-9-yl (MOX). In certain embodiments, Ti is a hydroxyl protecting group selected from acetyl, benzyl, t-butyldimethylsilyl, l-butyldiphenylsilyl and dimethoxytrityl wherein a more preferred hydroxyl protecting group Ti is 4,4'-dimethoxytrityl.

[00442] In certain embodiments, T2 is a reactive phosphorus group wherein preferred reactive phosphorus groups include di- / sopropylcyanoethoxy phosphoramidite and H-phosphonate. In certain embodiments, Ti is 4,4'-dimethoxytrityl and T2 is di- / sopropylcyanoethoxy phosphoramidite.

[00443] In certain embodiments, the compounds of the invention comprise at least one monomer of the formula: or from the formula: Petition 870260049905, dated 05 / 25 / 2026, page 157 / 728 144 / 335 or from the formula: in which Bx is a heterocyclic base moiety; T3 is H, a hydroxyl protecting group, a conjugate group attached, or an internucleoside linking group attached to a nucleoside, a nucleotide, an oligonucleoside, an oligonucleotide, a monomeric subunit, or an oligomeric compound; T4 is H, a hydroxyl protecting group, a conjugate group attached, or an internucleoside linking group attached to a nucleoside, a nucleotide, an oligonucleoside, an oligonucleotide, a monomeric subunit, or an oligomeric compound; wherein at least one of T3 and T4 is an internucleoside linking group attached to a nucleoside, a nucleotide, an oligonucleoside, an oligonucleotide, a monomeric subunit, or an oligomeric compound; and Z is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, substituted C1-C6 alkyl, substituted C2-C6 alkenyl, substituted C2-C6 alkynyl, acyl, substituted acyl, or substituted amide.

[00444] In some embodiments, each of the substituted groups is independently mono- or polysubstituted by optionally protected substituent groups independently selected from halogen, oxo, hydroxyl, OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 and CN, wherein each J1, J2 and J3 is, in Petition 870260049905, dated 05 / 25 / 2026, page 158 / 728 145 / 335 depending, H or C1-C6 alkyl, and X is O, S or NJ1.

[00445] In some embodiments, each of the substituted groups is independently mono- or polysubstituted by independently selected halogen, oxo, hydroxyl, OJ1, NJ1J2, SJ1, N3, OC(=X)J1 and NJ3C(=X)NJ1J2 substituent groups, where each J1, J2 and J3 is independently H or C1-Ce alkyl, and X is O or NJ1.

[00446] In certain such embodiments, at least one Z is a substituted C1-C1 alkyl or substituted O-C1 alkyl. In certain embodiments, each Z is independently a substituted C1-C1 alkyl or substituted C1-C1 alkyl. In certain embodiments, at least one Z is a C1-C1 alkyl. In certain embodiments, each Z is independently an O-C1 alkyl. In certain embodiments, at least one Z is methyl. In certain embodiments, each Z is methyl. In certain embodiments, at least one Z is ethyl. In certain embodiments, each Z is ethyl. In certain embodiments, at least one Z is a substituted O-C1 alkyl. In certain embodiments, each Z is independently a substituted C1-C1 alkyl. In certain embodiments, at least one Z is a substituted methyl. In certain embodiments, each Z is a substituted methyl. In certain embodiments, at least one Z is a substituted ethyl. In certain embodiments, each Z is a substituted ethyl.

[00447] In certain embodiments, at least one substituent group is a C1-C6 alkoxy (for example, at least one Z is a C1-C6 alkyl substituted by one or more C1-C6 alkoxy groups). In another embodiment, each substituent group is independently a C1-O6 alkoxy group (for example, each Z is independently an O6-O6 alkyl group substituted by one or more C1-C6 alkoxy groups).

[00448] In certain embodiments, at least one alkoxy substituent group Oi-Oθ is CH3O- (for example, at least one Z is CH3OCH2-). In another embodiment, each alkoxy substituent group Ci-Ce is CH3O (for example, each Z is CH3OCH2-). Petition 870260049905, dated 05 / 25 / 2026, p. 159 / 728 146 / 335

[00449] In certain embodiments, at least one substituent group is a halogen (for example, at least one Z is a C1-Ce alkyl substituted with one or more halogens). In certain embodiments, each substituent group is independently a halogen (for example, each Z is independently an O-Ce alkyl substituted with one or more halogens). In certain embodiments, at least one halogen substituent group is fluorine (for example, at least one Z is CH2FCH2-, CHF2CH2-, or CF3CH2-). In certain embodiments, each halo substituent group is fluorine (for example, each Z is independently CH2FCH2-, CHF2CH2-, or CF3CH2-).

[00450] In certain embodiments, at least one substituent group is hydroxyl (for example, at least one Z is a C1-C6 alkyl substituted with one or more hydroxyls). In certain embodiments, each substituent group is independently hydroxyl (for example, each Z is independently an O-Ce alkyl substituted with one or more hydroxyls). In certain embodiments, at least one Z is HOCH2-. In another embodiment, each Z is HOCH2-.

[00451] In certain embodiments, at least one Z is CH3-, CH3CH2-, CH2OCH3-, CH2F-, or HOCH2-. In certain embodiments, each Z is independently CH3-, CH3CH2-, CH2OCH3-, CH2F-, or HOCH2-.

[00452] In certain embodiments, at least one Z group is an Oi-Oθ alkyl substituted by one or more Xx, wherein each Xx is independently OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 or CN; wherein each J1, J2 and J3 is independently H or an Oi-Oθ alkyl, and X is O, S or NJ1. In another embodiment, at least one Z group is an Oi-Oθ alkyl substituted by one or more XX, wherein each Xx is independently a halo (e.g., fluorine), hydroxyl, alkoxy (e.g., CH3O-) or azide.

[00453] In certain modalities, each group Z is, independently Petition 870260049905, dated 05 / 25 / 2026, p. 160 / 728 147 / 335 te, Ci-Ce alkyl substituted by one or more Xx, where each Xx is independently OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 or CN; where each J1, J2 and J3 is independently H or Ci-Ce alkyl, and X is O, S or NJ1. In another embodiment, each Z group is independently O-Ce alkyl substituted by one or more XX, wherein each Xx is independently halo (e.g., fluorine), hydroxyl, alkoxy (e.g., CH3O-) or azide.

[00454] In certain embodiments, at least one Z group is -CH2XX, where Xx is OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 or CN; where each J1, J2 and J3 is independently H or C1-Ce alkyl, and X is O, S or NJ1. In certain embodiments, at least one Z group is -CH2XX, where Xx is halo (e.g., fluorine), hydroxyl, alkoxy (e.g., CH3O-) or azide.

[00455] In certain embodiments, each Z group is independently -CH2XX, where each Xx is independently OJ1, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2 or CN; where each J1, J2 and J3 is independently H or C1-Ce alkyl, and X is O, S or NJ1. In another embodiment, each Z group is independently CH2XX, where each Xx is independently halo (e.g., fluorine), hydroxyl, alkoxy (e.g., CH3O-) or azide.

[00456] In certain embodiments, at least one Z is CH3-. In another embodiment, every Z is CH3-.

[00457] In certain forms, the Z group of at least one monomer is in the (R) configuration represented by the formula: or by the formula: Petition 870260049905, dated 05 / 25 / 2026, p. 161 / 728 148 / 335 or by the formula:

[00458] In certain embodiments, the Z group of each monomer in the formula is in the (R) configuration.

[00459] In certain embodiments, the Z group of at least one monomer is in the (S) configuration represented by the formula: or by the formula: or by the formula:

[00460] In certain embodiments, the Z group of each monomer in the formula is in the (S) configuration.

[00461] In certain embodiments, T3 is H or a hydroxyl protecting group. In certain embodiments, T4 is H or a hydroxyl protecting group. In a further embodiment, T3 is an internucleoside linking group attached to a nucleoside, a nucleotide, or a molecular subunit. Petition 870260049905, dated 05 / 25 / 2026, page 162 / 728 149 / 335 numerical. In certain embodiments, T4 is an internucleoside linking group attached to a nucleoside, a nucleotide, or a monomeric subunit. In certain embodiments, T3 is an internucleoside linking group attached to an oligonucleoside or an oligonucleotide. In certain embodiments, T4 is an internucleoside linking group attached to an oligonucleoside or an oligonucleotide. In certain embodiments, T3 is an internucleoside linking group attached to an oligomeric compound. In certain embodiments, T4 is an internucleoside linking group attached to an oligomeric compound. In certain embodiments, at least one of T3 and T4 comprises an internucleoside linking group selected from phosphodiester or phosphorothioate.

[00462] In certain embodiments, the double-stranded RNA agent of the invention comprises at least one region of at least two contiguous monomers of the formula: or from the formula: or from the formula:

[00463] In certain such embodiments, LNAs include, but are not limited to, (A) α-L-Methylene-oxy(4'-CH2-O-2') LNA, (B) β-D-Methylene-oxy(4'-CH2-O-2') LNA, (C) Ethylene-oxy(4'-(CH2)2-O-2') LNA, Petition 870260049905, dated 05 / 25 / 2026, page 163 / 728 150 / 335 (D) Amino-oxy (4'-CH2-ON(R)-2') LNA and (E) Oxamino (4'CH2-N(R)-O-2') LNA, as illustrated below:

[00464] In certain embodiments, the double-stranded RNA agent of the invention comprises at least two regions of at least two contiguous monomers of the above formula: In certain embodiments, the double-stranded RNA agent of the invention comprises a gap motif. In certain embodiments, the double-stranded RNA agent of the invention comprises at least one region of about 8 to about 14 contiguous 3-D-2'-deoxyribofuranosyl nucleosides. In certain embodiments, the double-stranded RNA agent of the invention comprises Petition 870260049905, dated 05 / 25 / 2026, p. 164 / 728 151 / 335 at least one region of about 9 to about 12 contiguous 3-D-2'-deoxyribofuranosyl nucleosides.

[00465] In certain embodiments, the double-stranded RNA agent of the invention comprises at least one (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more) (S)-cEt monomer of the formula: S-cEt (C) J where Bx is the heterocyclic base portion.

[00466] In certain embodiments, the monomers include sugar mimetics. In certain such embodiments, a mimetic is used in place of the sugar or sugar-internucleoside linkage combination, and the nucleobase is retained for hybridization with a selected target. Representative examples of a sugar mimetic include, but are not limited to, cyclohexenyl or morpholino. Representative examples of a mimetic for a sugar-internucleoside linkage combination include, but are not limited to, peptide nucleic acids (PNA) and morpholino groups linked by uncharged achiral bonds. In some cases, a mimetic is used in place of the nucleobase. Representative nucleobase mimetics are well known in the art and include, but are not limited to, tricyclic phenoxazine analogs and universal bases (Berger et al., Nuc Acid Res. 2000, 28: 2911-14, incorporated herein by reference).Methods for synthesizing sugar, nucleoside, and nucleobase mimetics are well known to those skilled in the art. Nucleic acid modifications (intersugar interactions)

[00467] Linking groups that link monomers (including, but not limited to, nucleosides and modified nucleosides) are described herein. Petition 870260049905, dated 05 / 25 / 2026, page 165 / 728 152 / 335 fixed and unmodified) together, thus forming an oligomeric compound, for example, an oligonucleotide. Such linking groups are also referred to as intersugar linkages. The two main classes of linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing linkages include, but are not limited to, phosphodiesters (P=O), phosphotriesters, methylphosphonates, phosphoramidate and phosphorothioates (P=S). Representative non-phosphorus-containing linkage groups include, but are not limited to, methylenemethylimino (-CH2-N(CH3)-O-CH2-), thiodiester (-OC(O)-S-), thionocarbamate (-OC(O)(NH)-S-); siloxane (-OSi(H)2-O-); and N,N'-dimethylhydrazine (-CH2-N(CH3)-N(CH3)-). Modified linkages, compared to natural phosphodiester linkages, can be used to alter, typically increase, the nuclease resistance of oligonucleotides.In certain embodiments, linkages having a chiral atom can be prepared as racemic mixtures, as separate enantiomers. Representative chiral linkages include, but are not limited to, alkylphosphonates and phosphorothioates. Methods for preparing both phosphorus-containing and non-phosphorus-containing linkages are well known to those skilled in the art.

[00468] The phosphate group in the linking group can be modified by replacing one of the oxygens with a different substituent. A result of this modification can be increased resistance of the oligonucleotide to nucleolytic degradation. Examples of modified phosphate groups include phosphorothioates, phosphoroselenates, boranephosphates, boranephosphate esters, hydrogenphosphonates, phosphoramidates, alkyl or aryl phosphonates, and phosphotriesters. In some embodiments, one of the non-bridging phosphate oxygen atoms in the linkage can be substituted with any of the following: S, Se, BR3 (R is hydrogen, alkyl, aryl), C (i.e., an alkyl group, an aryl group, etc.), H, NR2 (R is hydrogen, optionally substituted alkyl, aryl). Petition 870260049905, dated 05 / 25 / 2026, page 166 / 728 153 / 335 or OR (R is optionally substituted alkyl or aryl). The phosphorus atom in an unmodified phosphate group is achiral. However, the substitution of one of the non-bridging oxygens by one of the atoms or groups of atoms above makes the phosphorus atom chiral; in other words, a phosphorus atom in a phosphate group modified in this way is a stereogenic center. The stereogenic phosphorus atom can have the R configuration (here Rp) or the S configuration (here Sp).

[00469] Phosphorodithioates have both non-bridged oxygens replaced by sulfur. The phosphorus center in phosphorodithioates is achiral, which prevents the formation of oligonucleotide diastereomers. Thus, while not wishing to be limited by theory, modifications to both non-bridged oxygens, which eliminate the chiral center, for example, phosphorodithioate formation, may be desirable insofar as they fail to produce mixtures of diastereomers. Thus, the non-bridged oxygens can independently be any one of O, S, Se, B, C, H, N, or OR (R is alkyl or aryl).

[00470] The phosphate linker can also be modified by replacing the bridging oxygen (i.e., the oxygen that links the phosphate to the sugar of the monomer) with nitrogen (bridging phosphorothioamidates), sulfur (bridging phosphorothioates), and carbon (bridging methylenephosphonates). The substitution can occur at either of the linking oxygens or at both of the linking oxygens. When the bridging oxygen is the 3' oxygen of a nucleoside, substitution with carbon is preferred. When the bridging oxygen is the 5' oxygen of a nucleoside, substitution with nitrogen is preferred.

[00471] Modified phosphate linkages where at least one of the oxygen atoms bonded to the phosphate has been substituted or the phosphate group has been replaced by a non-phosphorous group are also referred to as a non-differential phosphodiester intersugar linkage or non-differential linker. Petition 870260049905, dated 05 / 25 / 2026, page 167 / 728 154 / 335 phosphodiester.

[00472] In certain embodiments, the phosphate group may be replaced by non-phosphorus-containing connectors, for example, dephospho ligands. Dephospho ligands are also referred to as non-phosphodiester ligands herein. While not wishing to be limited by theory, it is believed that, since the charged phosphodiester group is the reaction center in nucleolytic degradation, its replacement by neutral structural mimetics should confer enhanced stability to nucleases. Again, while not wishing to be limited by theory, it may be desirable, in some embodiment, to introduce modifications in which the charged phosphate group is replaced by a neutral moiety.

[00473] Examples of moieties that can replace the phosphate group include, but are not limited to, amides (e.g., amide-3 (3'-CH2-C(=O)-N(H)-5') and amide-4 (3'-CH2-N(H)-C(=O)-5')), hydroxylamino, siloxane (dialkylsiloxane), carboxamide, carbonate, carboxymethyl, carbamate, carboxylate ester, thioether, ethylene oxide ligand, sulfide, sulfonate, sulfonamide, sulfonate ester, thioformacetal (3'-SCH2-O-5'), formacetal (3'-O-CH2-O-5'), oxime, methyleneimino, methylenecarbonylamino, methylenemethylimino (MMI, 3'-CH2-N(CH3)-O-5'), methylenehydrazo, methylenedimethylhydrazone, methyleneoxymethylimino, (C3-O-C5') ethers, (C3'-S-C5') thioethers, thioacetamido (C3'-N(H)-C(=O)-CH2S-C5', C3'-OP(O)-O-SS-C5', C3'-CH2-NH-NH-C5', 3'-NHP(O)(OCH3)0-5' and 3'-NHP(O)(OCH3)-O-5' and non-ionic linkages containing N, O, S and mixed CH2 component parts. See, for example, 'Carbohydrate Modifications in Antisense Research', YS Sanghvi and PD Cook Eds. ACS Symposium Series 580; Chapters 3 and 4 (pp. 40-65).Preferred embodiments include methylenemethylimino (MMI), methylenecarbonylamino, amides, carbamate, and ethylene oxide binder.

[00474] A person versed in the technique is well aware that in Petition 870260049905, dated 05 / 25 / 2026, page 168 / 728 155 / 335 In certain cases, the substitution of a non-bridged oxygen can lead to intensified cleavage of the intersugar bond by the neighboring 2'-OH. Thus, in many cases, a modification of a non-bridged oxygen may require modification of 2'-OH, for example, a modification that does not participate in the cleavage of the neighboring intersugar bond, for example, arabinose sugar, 2'-O-alkyl, 2'-F, LNA and ENA.

[00475] Preferred intersugar linkages other than phosphodiester include phosphorothioates, phosphorothioates with at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more enantiomeric excess of the sp isomer, phosphorothioates with at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more enantiomeric excess of the rp isomer, phosphorodithioates, phosphotriesters, aminoalkylphosphotrioesters, alkylphosphonates (e.g., methylphosphonate), selenophosphates, phosphoramidates (e.g., N-alkylphosphoramidate) and boranephosphonates.

[00476] In some embodiments, the double-stranded RNA agent of the invention comprises at least one (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more and even including all) modified or different phosphodiester linkages. In some embodiments, the double-stranded RNA agent of the invention comprises at least one (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more and even including all) phosphorothioate linkages.

[00477] The double-stranded RNA agent of the inventions can also be constructed in which the phosphate linker and sugar are replaced by nuclease-resistant nucleoside or nucleotide substitutes. While not wishing to be limited by theory, it is believed that the absence of a repeatedly loaded backbone diminishes binding to proteins that recognize polyanions (e.g., nucleases). Again, while not wishing to be limited by theory, it may be desirable, in some embodiments, to introduce alterations Petition 870260049905, dated 05 / 25 / 2026, page 169 / 728 156 / 335 tions in which the bases are linked by a neutral substituent backbone. Examples include morpholino, cyclobutyl, pyrrolidine, peptide nucleic acid (PNA), aminoethylglycyl PNA (aegPNA), and PNA with backbone extension (bepPNA) nucleoside substitutes. A preferred substitute is a PNA substitute.

[00478] The double-stranded RNA agent of the invention described herein may contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers and other stereoisomeric configurations which may be defined, in terms of absolute stereochemistry, as (R) or (S), such as for sugar anomers, or as (D) or (L) such as for amino acids et al. All such possible isomers, as well as their racemic and optically pure forms, are included in the double-stranded RNA agent of the invention provided herein. Nucleic acid modifications (terminal modifications)

[00479] In some embodiments, the double-stranded iRNA agent additionally comprises a phosphate or phosphate mimetic at the 5' end of the antisense strand. In one embodiment, the phosphate mimetic is a 5'-vinyl phosphonate (VP).

[00480] In some embodiments, the 5' end of the antisense strand of the double-stranded iRNA agent does not contain a 5'-vinyl phosphonate (VP).

[00481] The ends of the RNA agent of the invention can be modified. Such modifications can be at one end or both ends. For example, the 3' and / or 5' ends of an RNA can be conjugated to other functional molecular entities such as labeling moieties, for example, fluorophores (e.g., pyrene, TAMRA, fluorescein, Cy3 or Cy5 dyes) or protecting groups (based, for example, on sulfur, silicon, boron or ester). Functional molecular entities can be attached to the sugar via a phosphate group and / or a linker. The terminal atom Petition 870260049905, dated 05 / 25 / 2026, p. 170 / 728 157 / 335 of the ligand can connect to or replace the linking atom of the phosphate group or the C-3' or C-5' O, N, S, or C group of the sugar. Alternatively, the ligand can connect to or replace the terminal atom of a nucleotide substitute (e.g., PNAs).

[00482] When a phosphate-ligand / phosphate network of functional molecular entities is interposed between two strands of a double-stranded oligomeric compound, this network can replace a hairpin loop in a hairpin-type oligomeric compound.

[00483] Terminal modifications useful for modulating activity include modification of the 5' end of iRNAs with phosphate or phosphate analogs. In certain embodiments, the 5' end of an iRNA is phosphorylated or includes a phosphoryl analog. Exemplary 5'-phosphate modifications include those compatible with RISC-mediated gene silencing. 5'-terminal modifications may also be useful in stimulating or inhibiting an individual's immune system. In some embodiments, the 5' end of the oligomeric compound comprises the modification- tion, wherein W, X, and Y are each independently selected from the group consisting of O, OR (R is hydrogen, alkyl, aryl), S, Se, BR3 (R is hydrogen, alkyl, aryl), BH3·, C (i.e., an alkyl group, an aryl group, etc.), H, NR2 (R is hydrogen, alkyl, aryl), or OR (R is hydrogen, alkyl, or aryl); A and Z are each independently selected for each missing occurrence, are O, S, CH2, NR (R is hydrogen, alkyl, aryl), or optionally substituted alkylene, wherein the alkylene skeleton may comprise one or more of O, S, SS, and NR (R is hydrogen, alkyl, aryl) internally and / or at the end; n is 0-2. In some embodiments, n is 1 or 2. It is understood that A is replacing the oxygen bonded to carbon 5' of the sugar. Petition 870260049905, dated 05 / 25 / 2026, page 171 / 728 158 / 335 When n is 0, W and Y together with the P to which they are attached can form an optionally substituted 5-8 membered heterocycle, wherein W and Y are each independently O, S, NR', or alkylene. Preferably, the heterocycle is substituted by an aryl or heteroaryl. In some embodiments, one or both of the hydrogens at C5' of the 5'-terminal nucleotides are substituted by a halogen, for example, F.

[00484] 5'-Exemplary modifications include, but are not limited to, 5'-monophosphate ((HO)2(O)PO-5'); 5'-diphosphate ((HO)2(O)PO-P(HO)(O)-O-5'); 5'-triphosphate ((HO)2(O)PO-(HO)(O)POP(HO)(O)-O5'); 5'-monothiophosphate (phosphorothioate; (HO)2(S)PO-5'); 5'-monodithiophosphate (phosphorodithioate; (HO)(HS)(S)PO-5'), 5'-phosphorothiolate ((HO)2(O)PS-5'); 5'-alpha-thiotriphosphate; 5'-beta-thiotriphosphate; 5'-gamma-thiotriphosphate; 5'-phosphoramidates ((HO)2(O)P-NH-5', (HO)(NH2)(O)PO-5'). Other 5'-modifications include 5'-alkylphosphonates (R(OH)(O)PO-5', R=alkyl, e.g., methyl, ethyl, Isopropyl, propyl, etc...), 5'-alkyl etherphosphonates (R(OH)(O)PO-5', R=alkyl ether, e.g., methoxymethyl (CH2OMe), ethoxymethyl, etc...).Other exemplary 5'-modifications include where Z is optionally substituted alkyl at least once, for example, ((HO)2(X)PO[-(CH2)aOP(X)(OH)-O]b5', ((HO)2(X)PO[-(CH2)aP(X)(OH)-O]b- 5', ((HO)2(X)P-[-(CH2)a-OP(X)(OH)-O]b- 5'; dialkyl-terminated phosphates and phosphate mimetics: HO[-(CH2)aOP(X)(OH)-O]b- 5', H2N[-(CH2)aOP(X)(OH)-O]b5', H[-(CH2)aOP(X)(OH)-O]b- 5', Me2N[-(CH2)aOP(X)(OH)-O]b- 5', HO[-(CH2)aP(X)(OH)-O]b- 5', H2N[-(CH2)aP(X)(OH)-O]b- 5', H[-(CH2)aP(X)(OH)-O]b- 5', Me2N[-(CH2)aP(X)(OH)-O]b- 5', where a and b are each independently 1-10. and / or sulfur by BH3, BH3· and / or Se.

[00485] Terminal modifications can also be useful for monitoring distribution and, in such cases, preferential groups to Petition 870260049905, dated 05 / 25 / 2026, p. 172 / 728 159 / 335 to be added include fluorophores, for example, fluorescein or an Alexa dye, for example, Alexa 488. Terminal modifications can also be useful for enhancing uptake; useful modifications for this include targeting ligands. Terminal modifications can also be useful for crosslinking one oligonucleotide to another moiety; useful modifications for this include mitomycin C, psoralen and their derivatives. Thermally Destabilizing Modifications

[00486] The compounds of the invention, such as iRNAs or dsRNA agents, can be optimized for RNA interference by increasing the propensity of the iRNA duplex to dissociate or fuse (decreasing the free energy of association of the duplex) by introducing a thermally destabilizing modification in the sense strand at a location opposite to the seed region of the antisense strand (i.e., at positions 2-8 of the 5' end of the antisense strand). This modification can increase the propensity of the duplex to dissociate or fuse in the seed region of the antisense strand.

[00487] Thermally destabilizing modifications may include abasic modification; mismatch with the opposite nucleotide on the opposite strand; and sugar modification such as 2'-deoxy or acyclic nucleotide modification, for example, unblocked nucleic acids (UNA) or glycerol nucleic acid (GNA).

[00488] Basic modifications exemplified are:

[00489] Examples of sugar modifications are: Petition 870260049905, dated 05 / 25 / 2026, p. 173 / 728 160 / 335 o unlocked nucleic acid glycol 2 —deoxy nucleic acid R= H, OH, O-alkyl R=H, OH, O-alkyl

[00490] The term acidic nucleotide refers to any nucleotide having an acidic ribose sugar, for example, where any of the linkages between the ribose carbons (e.g., C1-C2', C2'-C3', C3'-C4', O4-O4', or C1'-O4') is absent and / or at least one of the ribose carbons or oxygen (e.g., OΓ, C2', C3', O4', or O4') is independently or in combination absent from the nucleotide. In some embodiments, the acidic nucleotide i It is a modified or unmodified nucleobase, R1 and R2 are independently H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar. The term UNA refers to unblocked acidic nucleic acid, in which any of the sugar bonds have been removed, forming an unblocked sugar residue. In one example, UNA also encompasses monomers with O1-O4' bonds being removed (i.e., the carbon-oxygen-carbon covalent bond between carbons O1' and C4'). In another example, the C2'-C3' linkage (that is, the carbon-carbon covalent bond between carbons O2' and O3') of the sugar is removed (see Mikhailov et al., Tetrahedron Letters, 26 (17): 2059 (1985); and Fluiter et al., Mol. Biosyst., 10: 1039 (2009), which are thus incorporated by ref Petition 870260049905, dated 05 / 25 / 2026, p. 174 / 728 161 / 335 (reference in its entirety). The acidic derivative provides greater flexibility of the backbone without affecting Watson-Crick pairings. The acidic nucleotide can be linked via 2'-5' or 3'-5' linkages.

[00491] The term GNA refers to glycol nucleic acid, which is a polymer similar to DNA or RNA but differing in the composition of its backbone in that it is composed of repeating glycerol units linked by phosphodiester bonds: O (R)-GNA

[00492] Thermally destabilizing modifications can be mismatches (i.e., non-complementary base pairs) between the thermally destabilizing nucleotide and the opposite nucleotide on the opposite strand within the dsRNA duplex. Exemplary mismatch base pairs include G:G, G:A, G:U, G:T, A:A, A:C, C:C, C:U, C:T, U:U, T:T, U:T, or a combination thereof. Other mismatch base pairings known in the art are also suitable for the present invention. A mismatch can occur between nucleotides that are naturally occurring nucleotides or modified nucleotides, i.e., mismatch base pairing can occur between the nucleobases of the respective nucleotides independently of modifications to the ribose sugars of the nucleotides. In certain embodiments, the compounds of the invention, such as siRNA or iRNA agent, contain at least one nucleobase in the mismatched pairing that is a 2'-deoxy nucleobase; Petition 870260049905, dated 05 / 25 / 2026, page 175 / 728 162 / 335 for example, a 2-deoxy nucleobase is on the sense strand.

[00493] Further examples of abasic nucleotides, acyclic nucleotide modifications (including UNA and GNA) and mismatch modifications have been described in detail in WO 2011 / 133876, which is incorporated herein by reference in its entirety.

[00494] Thermally destabilizing modifications may also include universal bases with reduced or abolished capacity to form hydrogen bonds with opposing bases and phosphate modifications.

[00495] Nucleobase modifications with impaired or completely abolished ability to form hydrogen bonds with bases on the opposite strand were evaluated for destabilization of the central region of the dsRNA duplex as described in WO 2010 / 0011895, which is incorporated herein by reference in its entirety. Examples of nucleobase modifications are: THE inosine nebularine 2-aminopurine difluorotoluene 5-nitroindole 3-nitropyrrole 4-Fluoro-6- 4-Methylbenzimidazole methylbenzimidazole

[00496] Exemplary phosphate modifications known to decrease the thermal stability of dsRNA duplexes compared to natural phosphodiester bonds are: ό 0 ό ό ό ό O=P—SH O=P-CHj 1 o=p-ch2-cooh O=PR O=P-NH-R O=POR 0 o ό ό ό ό R=alkyl

[00497] In some embodiments, the compounds of the invention po Petition 870260049905, dated 05 / 25 / 2026, p. 176 / 728 163 / 335 dem comprise 2-5' linkages (with 2'-H, 2'-OH and 2'-OMe and with P=O or P=S). For example, modifications of the 2-5' linkages can be used to promote resistance to nucleases or to inhibit the binding of the sense strand to the antisense strand, or they can be used at the 5' end of the sense strand to prevent activation of the sense strand by RISC.

[00498] In another embodiment, the compounds of the invention may comprise L sugars (e.g., L-ribose, L-arabinose with 2'H, 2'-OH and 2'-OMe). For example, these L-sugar modifications may be used to promote resistance to nucleases or to inhibit the binding of the sense strand to the antisense strand or may be used at the 5' end of the sense strand to prevent activation of the sense strand by RISC.

[00499] In one embodiment, the RNA agent of the invention is conjugated to a ligand via a carrier, wherein the carrier may be a cyclic group or an acyclic group; preferably, the cyclic group is selected from pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl and decalin; preferably, the acyclic group is selected from a serinol skeleton or a diethanolamine skeleton.

[00500] In some embodiments, at least one strand of the RNA agent of the invention described herein is phosphorylated at 5' or includes a 5'-terminal phosphoryl analog. 5'-phosphate modifications include those that are compatible with RISC-mediated gene silencing. Suitable modifications include: 5'-monophosphate ((HO)2(O)PO-5'); 5'-diphosphate ((HO)2(O)POP(HO)(O)-O-5'); 5'-triphosphate ((HO)2(O)PO-(HO)(O)POP(HO)(O)-O-5'); 5'-guanosine cap (7-methylated or unmethylated) (7m-GO-5'-(HO)(O)PO(HO)(O)POP(HO)(O)-O-5'); 5'-adenosine cap (Appp) and any Petition 870260049905, dated 05 / 25 / 2026, page 177 / 728 164 / 335 Modified or unmodified nucleotide cap structure (N-O5'-(HO)(O)PO-(HO)(O)POP(HO)(O)-O-5'); 5'-monothiophosphate (phosphorothioate; (HO)2(S)PO-5'); 5'-monodithiophosphate (phosphorodithioate; (HO)(HS)(S)PO-5'), 5'-phosphorothiolate ((HO)2(O)PS-5'); any additional combination of oxygen / sulfur substituted monophosphate, diphosphate and triphosphates (e.g., 5'-alpha-thiotriphosphate, 5'-gamma-thiotriphosphate, etc.), 5'-phosphoramidates ((HO)2(O)P-NH-5', (HO)(NH2)(O)P0-5'), 5'-alkylphosphonates (R=alkyl=methyl, ethyl, / sopropyl, propyl, etc., e.g., RP(OH)(O)-O-5'-, 5'-alkenylphosphonates (i.e., vinyl, substituted vinyl), (OH)2(O)P-5'-CH2-), 5'-alkyletherphosphonates (R=alkylether=methoxymethyl (MeOCH2-), ethoxymethyl, etc., e.g., RP(OH)(O)-O-5'-). Target genes

[00501] Without limitation, target genes for siRNAs include, but are not limited to, genes promoting unwanted cell proliferation, growth factor genes, growth factor receptor genes, genes expressing kinases, an adaptor protein gene, a gene encoding a molecule of the G protein superfamily, a gene encoding a transcription factor, a gene mediating angiogenesis, a viral gene, a gene required for viral replication, a cellular gene mediating viral function, a gene from a bacterial pathogen, a gene from an amoebic pathogen, a gene from a parasitic pathogen, a gene from a fungal pathogen, a gene mediating an unwanted immune response, a gene mediating pain processing, a gene mediating a neurological disease, an allele gene found in cells characterized by loss of heterozygosity, or an allele gene of a polymorphic gene.

[00502] Specific exemplary target genes for siRNAs include, but are not limited to, PCSK-9, ApoC3, AT3, AGT, ALAS1, TMPR, HAO1, AGT, C5, CCR-5, PDGF beta gene; gene Petition 870260049905, dated 05 / 25 / 2026, p. 178 / 728 165 / 335 Erb-B, Src gene; CRK gene; GRB2 gene; RAS gene; MEKK gene; JNK gene; RAF gene; Erk1 / 2 gene; PCNA gene (p21); MYB gene; cMYC gene; JUN gene; FOS gene; BCL-2 gene; Cyclin D gene; VEGF gene; EGFR gene; Cyclin A gene; Cyclin E gene; WNT1 gene; beta-catenin gene; c-MET gene; PKC gene; NFKB gene; STAT3 gene; survivin gene; Her2 / Neu gene; topoisomerase I gene; topoisomerase II alpha gene; p73 gene; p21(WAF1 / CIP1) gene, p27(KIP1) gene; PPM1D gene; caveolin I gene; MIB I gene; MTAI gene; M68 gene; tumor suppressor genes; p53 gene; DN-p63 gene; tumor suppressor gene pRb; tumor suppressor gene APC1; tumor suppressor gene BRCA1; tumor suppressor gene PTEN; MLL fusion genes, for example, MLL-AF9, BCR / ABL fusion gene; TEL / AML1 fusion gene; EWS / FLI1 fusion gene; TLS / FUS1 fusion gene; PAX3 / FKHR fusion gene; AML1 / ETO fusion gene; alpha v-integrin gene; Flt-1 receptor gene; tubulin gene; Human Papillomavirus gene,a gene required for the replication of the Human Papillomavirus, a gene for the Human Immunodeficiency Virus, a gene required for the replication of the Human Immunodeficiency Virus, a gene for the Hepatitis A Virus, a gene required for the replication of the Hepatitis A Virus, a gene required for the replication of the Hepatitis B Virus, a gene required for the replication of the Hepatitis B Virus, a gene required for the replication of the Hepatitis C Virus, a gene required for the replication of the Hepatitis C Virus, a gene required for the replication of the Hepatitis D Virus, a gene required for the replication of the Hepatitis D Virus, a gene required for the replication of the Hepatitis E Virus, a gene required for the replication of the Hepatitis E Virus, a gene required for the replication of the Hepatitis F Virus, a gene required for the replication of the Hepatitis F Virus, a gene required for the replication of the Hepatitis G Virus, a gene required for the replication of the Hepatitis G Virus, a gene required for the replication of the Hepatitis G Virus H, urn gene required for the replication of the Hepatitis H Virus, gene of the Respiratory Syncytial Virus, urn gene that is required for replication, Petition 870260049905, dated 05 / 25 / 2026, p. 179 / 728 166 / 335 Respiratory Syncytial Virus gene, Herpes Simplex virus gene, a gene required for the replication of Herpes Simplex virus, Cytomegalovirus herpes gene, a gene required for the replication of Cytomegalovirus herpes, Epstein-Barr virus herpes gene, a gene required for the replication of Epstein-Barr virus herpes, Kaposi's Sarcoma-associated Herpesvirus gene, a gene required for the replication of Kaposi's Sarcoma-associated Herpesvirus, JC virus gene, a human gene required for the replication of JC virus, myxovirus gene, a gene required for the replication of the myxovirus gene, rhinovirus gene, a gene required for the replication of rhinovirus, coronavirus gene, a gene required for the replication of coronavirus, West Nile virus gene, a gene that is required for the replication of the West Nile Virus, gene for St. Louis Encephalitis,a gene that is necessary for the replication of St. Louis encephalitis, a gene of tick-borne encephalitis virus, a gene that is required for the replication of tick-borne encephalitis virus, a gene of Murray Valley encephalitis virus, a gene that is required for the replication of Murray Valley encephalitis virus, a gene of dengue virus, a gene that is required for the replication of dengue virus gene, a gene of Simian Virus 40, a gene required for the replication of Simian Virus 40, a gene of Human T-cell Lymphotropic Virus, a gene required for the replication of Human T-cell Lymphotropic Virus, a gene of Moloney Murine Leukemia Virus, a gene that is required for the replication of Moloney Murine Leukemia Virus, a gene of encephalomyocarditis virus, a gene that is required for the replication of the virus encephalomyocarditis, measles virus gene, a gene that is necessary for measles virus replication,Varicella zoster virus gene, a gene that is necessary for the replication of the varicella zoster virus, gene, Petition 870260049905, dated 05 / 25 / 2026, p. 180 / 728 167 / 335 of adenovirus, a gene that is required for adenovirus replication, yellow fever virus gene, a gene required for yellow fever virus replication, poliovirus gene, a gene that is required for poliovirus replication, poxvirus gene, a gene that is required for poxvirus replication, Plasmodium gene, a gene that is required for Plasmodium gene replication, Mycobacterium ulcerans gene, a gene that is required for Mycobacterium ulcerans replication, Mycobacterium tuberculosis gene, a gene that is required for Mycobacterium tuberculosis replication, Mycobacterium leprae gene, a gene that is required for Mycobacterium leprae replication, Staphylococcus aureus gene, a gene that is required for Staphylococcus aureus replication, Streptococcus pneumoniae gene, a gene that is required for replication of Streptococcus pneumoniae, Streptococcus pyogenes gene,a gene required for the replication of Streptococcus pyogenes, a gene for Chlamydia pneumoniae, a gene required for the replication of Chlamydia pneumoniae, a gene for Mycoplasma pneumoniae, a gene required for the replication of Mycoplasma pneumoniae, an integrin gene, a selectin gene, a complement system gene, a chemokine gene, a chemokine receptor gene, a GCSF gene, a Gro1 gene, a Gro2 gene, a Gro3 gene, a PF4 gene, a MIG gene, a proplatelet basic protein gene, a MIP11 gene, a MIP-1J gene, a RANTES gene, a MCP-1 gene, a MCP-2 gene, a MCP-3 gene, a CMBKR1 gene, a CMBKR2 gene, a CMBKR3 gene, a CMBKR5v gene, an AIF-1 gene, an I-309 gene, a gene for a component of an ion channel, a gene for a receptor of neurotransmitter, a gene for a neurotransmitter ligand, amyloid family gene, presenilin gene, HD gene, DRPLA gene, SCA1 gene, SCA2 gene, MJD1 gene, CACNL1A4 gene, SCA7 gene, SCA8 gene, allele gene found in cells with loss of heterozygosity (LOH), a gene, Petition 870260049905, dated 05 / 25 / 2026, page 181 / 728 168 / 335 allele of a polymorphic gene and its combinations.

[00503] Loss of heterozygosity (LOH) can result in hemizygosity for the sequence, for example, genes, in the LOH region. This can result in a significant genetic difference between normal cells and diseased cells, for example, cancerous cells, and provides a useful difference between normal cells and diseased cells, for example, cancerous cells. This difference can arise because a gene or other sequence is heterozygous in doublet cells, but is hemizygous in cells having LOH. LOH regions will often include a gene, the loss of which promotes unwanted proliferation, for example, a tumor suppressor gene, and other sequences including, for example, other genes, in some cases a gene that is essential for normal function, for example, growth. The methods of the invention are based, in part, on the specific modulation of an allele of an essential gene with a composition of the invention.

[00504] In certain embodiments, the invention provides a double-stranded iRNA agent of the invention that modulates a micro-RNA. Central Nervous System (CNS) targeting

[00505] In some embodiments, the invention provides a double-stranded RNA agent targeting APR for Early-Onset Familial Alzheimer's Disease, ATXN2 for Spinocerebellar Ataxia 2 and ALS, and C9orf72 for Amyotrophic Lateral Sclerosis and Frontotemporal Dementia.

[00506] In some embodiments, the invention provides a double-stranded RNA agent targeting TARDBP for ALS, MAPT (Tau) for Frontotemporal Dementia and HTT for Huntington's Disease.

[00507] In some embodiments, the invention provides a double-stranded RNA agent targeting SNCA for Parkinson's Disease, FUS for ALS, ATXN3 for Spinocerebellar Ataxia 3, ATXN1 Petition 870260049905, dated 05 / 25 / 2026, p. 182 / 728 169 / 335 for SCA1, genes for SCA7 and SCA8, ATN1 for DRPLA, MeCP2 for XLMR, PRNP for Prion Diseases, recessive CNS disorders: Lafora Disease, DMPK for DM1 (CNS and Skeletal Muscle) and TTR for hATTR (CNS, ocular and systemic).

[00508] Spinocerebellar ataxia is a hereditary dysfunction of brain function. The predominantly hereditary forms of spinocerebellar ataxias, such as SCA1-8, are devastating dysfunctions without disease-modifying therapy. Exemplary targets include SCA2, SCA3, and SCA1. ATXN2 routing to SCA2

[00509] Spinocerebellar Ataxia 2 (SCA2), a progressive ataxia, is the second most common SCA. Another disease associated with this target is amyotrophic lateral sclerosis (ALS). These diseases are debilitating and ultimately fatal without disease-modifying therapy. The prevalence of SCA is 2-6 per 100,000 people; ATXN2 affects 15% of the population with SCA globally and many more populations with SCA in some countries, especially in Cuba (40 per 100,000 people). Targeting ATXN2 may be highly effective through human molecular genetics; for example, expansion of the CAG repeat encoding ATXN2 has been discovered in familial and sporadic SCA and ALS, in tissues such as the spinal cord, brainstem, or cerebellum. The mechanism of this targeting may be because the expansion of autosomal dominant CAG encoding ATXN2 causes the expression of a misfolded, toxic protein and death of Purkinje cells and neurons.Efficacy was demonstrated by 70% silencing (KD) of ATXN2 mRNA; and KD mATXN2 POC mice were shown. Regarding safety, knockout (KO) mice for mATXN2 were reported as healthy. Possible diagnosis includes family history; genetic testing; or initial symptoms. Biomarkers that can be used include, for example... Petition 870260049905, dated 05 / 25 / 2026, page 183 / 728 170 / 335 example, CAG mRNA in CSF and peptide repeat proteins. Targeting of ATXN3 to SCA3.

[00510] Spinocerebellar Ataxia 3 (SCA3), a progressive ataxia, is the most common SOA globally. This disease is debilitating and ultimately lethal without disease-modifying therapy. It is the most common cause of SCA, and the prevalence of SCA is 2-6 per 100,000 people; ATXN3 causes 21% of the population with SCA in the US and much more in Europe, especially in Portugal. Targeting ATXN3 may be excellent through human molecular genetics, for example, the expansion of the ATXN3-coding CAG repeat has been discovered in familial and sporadic SCA, in tissues such as the spinal cord, brainstem, or cerebellum. The mechanism of this targeting may be because the autosomal dominant CAG expansion encoding ATXN3 causes expression of a misfolded, toxic protein, death of Purkinje cells and neurons. Efficacy has been shown by 70% KD of ATXN3 mRNA; and KD mATXN3 POC mice were shown.Regarding safety, mATXN3 KO mice have been reported as healthy. Possible diagnoses include family history; genetic testing; or initial symptoms. Biomarkers that can be used include, for example, CAG mRNA in the CSF and peptide repeat proteins. ATXN1 routing to SCA1

[00511] Spinocerebellar Ataxia 1 (SCA1), a progressive ataxia, is the first SCA gene discovered in 1993. This disease is debilitating and ultimately fatal without disease-modifying therapy. The prevalence of SCA is 2-6 per 100,000 people; ATXN1 causes 6% of the population with SCA in the US and globally, and much more in some countries (25% in Japan), especially in Poland (64%) and Siberia (100%). Targeting ATXN1 may be excellent through human molecular genetics, for example, expansion Petition 870260049905, dated 05 / 25 / 2026, page 184 / 728 The 171 / 335 CAG repeat encoding ATXN1 has been discovered in familial and sporadic SCA, in tissues such as the spinal cord, brainstem, or cerebellum. The mechanism of this targeting may be because the expansion of the autosomal dominant CAG encoding ATXN1 causes the expression of a misfolded, toxic protein, leading to Purkinje cell and neuronal death. Efficacy has been shown by 70% kDa of ATXN1 mRNA; and mATXNI POC mice have been demonstrated. Regarding safety, mATXNI KO mice have been reported as healthy. Possible diagnosis includes family history; genetic testing; or initial symptoms. Biomarkers that can be used include, for example, CAG mRNA in the CSF and peptide repeat proteins. ATXN7 to SCA7 routing

[00512] Spinocerebellar Ataxia 7 (SCA7) causes progressive ataxia and retinal degeneration. This disease is a debilitating retinal and cerebellar dysfunction and ultimately lethal without disease-modifying therapy. The prevalence of SCA is 2-6 per 100,000 people; ATXN7 causes 5% of the population with SCA globally, and much more in some countries, especially in South Africa. Targeting of ATXN7 may be excellent through human molecular genetics, for example, the expansion of the CAG repeat encoding ATXN7 has been discovered in familial and sporadic SCA, in tissues such as spinal cord, brainstem, cerebellum or retina. The mechanism of this targeting may be because the autosomal dominant CAG expansion encoding ATXN1 causes expression of a toxic misfolded protein, inciting cone-rod dystrophy, Purkinje cell lethality and neuronal lethality. Efficacy was demonstrated by 70% kDa of ATXN1 mRNA via intrathecal (IT) and intravitreal (IVT) administration.Possible diagnoses include family history; genetic testing; or early symptoms. Biomarkers that can be used include, for example... Petition 870260049905, dated 05 / 25 / 2026, page 185 / 728 172 / 335 pIO, CAG mRNA in CSF and peptide repeat proteins. ATXN8 targeting to SCA8

[00513] Spinocerebellar Ataxia 8 (SCA8), a progressive neurodegenerative ataxia, is caused by CTG repeat expansion in ATXN8. This disease is debilitating and ultimately fatal without disease-modifying therapy. The prevalence of SCA is 2-6 per 100,000 people; ATXN8 causes 3% of the population with SCA globally, and much more in some countries, especially Finland. Targeting ATXN8 may be excellent through human molecular genetics, for example, the expansion of the CTG repeat encoding ATXN8 has been discovered in familial and sporadic SCA, in ti...

Claims

CLAIMS 1. Double-stranded RNA agent, characterized in that it comprises: an antisense strand that is complementary to a target gene; a sense strand that is complementary to said antisense strand; and one or more lipophilic portions conjugated to one or more internal positions in at least one strand, optionally via a linker or carrier, wherein one or more lipophilic portions comprise a saturated or unsaturated C4-C18 hydrocarbon chain and an optional functional group selected from the group consisting of hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide and alkyne; wherein one or more lipophilic fractions are conjugated to one or more of the following internal positions: positions 4-8 and 13-18 in the sense strand and positions 6-10 and 15-18 in the antisense strand, counting from the 5' end of each strand;and wherein the double-stranded RNA agent is characterized by one of the following features: (i) the antisense strand comprises a phosphate or phosphate analog at the 5' end of the antisense strand; or (ii) the sense strand contains a motif of three 2'-F modifications at three consecutive nucleotides, at positions 7 to 15 from the 5' end of the sense strand, the sense strand contains fewer than ten 2'-F modifications, and the antisense strand contains two blocks of two phosphorothioate or methylphosphonate internucleotide bonds separated by 16 to 18 phosphate internucleotide bonds; or (iii) the sense strand contains fewer than ten 2'-F modifications, and the double-stranded RNA agent comprises at least 6 Petition 870260049905, dated 05 / 25 / 2026, p. 350 / 728 2 / 6 internucleotide bonds of phosphorothioate or methylphosphonate.

2. Double-stranded iRNA agent according to claim 1, characterized in that the antisense strand comprises a phosphate or a phosphate analog at the 5' end of the antisense strand.

3. Double-stranded RNA agent according to claim 2, characterized in that the phosphate analog is a 5'-vinylphosphonate (VP).

4. Double-stranded iRNA agent according to claim 1, characterized in that: the sense strand contains a motif of three 2'-F modifications on three consecutive nucleotides, at positions 7 to 15 from the 5' end of the sense strand; the antisense strand contains fewer than ten 2'-F modifications; and the antisense strand contains two blocks of two phosphorothioate or methylphosphonate internucleotide bonds, separated by 16 to 18 phosphate internucleotide bonds.

5. Double-stranded RNA agent according to claim 1, characterized in that: the sense strand contains fewer than ten 2'-F modifications, and the double-stranded RNA agent comprises at least 6 phosphorothioate or methylphosphonate internucleotide linkages.

6. Double-stranded iRNA agent according to any one of claims 1 to 5, characterized in that one or more lipophilic moieties are conjugated to one or more of the following internal positions: positions 5, 6, 7, 15 and 17 on the sense strand and positions 15 and 17 on the antisense strand, counting from the 5' end of each strand.

7. Double-stranded RNA agent according to any one of claims 1 to 5, characterized in that the sense and antisense strands are each 15 to 30 nucleotides long.

8. Double-stranded RNA agent according to any one of claims 1 to 5, characterized in that the sense and antisense strands each have a length of 19 to 25 nucleotides.

9. Double-stranded RNA agent according to any one of claims 1 to 5, characterized in that the sense and antisense strands each have a length of 21 to 23 nucleotides.

10. Double-stranded RNA agent according to claim 9, characterized in that the sense strand is 21 nucleotides long and the antisense strands are 23 nucleotides long, wherein the strands form a double-stranded region of 21 consecutive base pairs with a protruding 3' single-stranded end of 2 nucleotides.

11. Double-stranded RNA agent according to any one of claims 1 to 10, characterized in that the lipophilic portion contains a saturated or unsaturated Ce-Cw hydrocarbon chain.

12. Double-stranded RNA agent according to claim 11, characterized in that the lipophilic portion contains a saturated or unsaturated Cw hydrocarbon chain.

13. Double-stranded RNA agent according to any one of claims 1 to 12, characterized in that the lipophilic portion is conjugated by means of a carrier that replaces one or more nucleotides at the internal position(s).

14. Double-stranded RNA agent according to claim 13, characterized in that the carrier is a cyclic group selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, Petition 870260049905, 05 / 25 / 2026, p. 352 / 728 4 / 6 [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydroforanyl and decalinyl; or is an acyclic moiety based on a serinol structure or a diethanolamine structure.

15. Double-stranded RNA agent according to any one of claims 1 to 14, characterized in that the lipophilic portion is conjugated to the double-stranded RNA agent by means of a linker containing an ether, thioether, urea, carbonate, amine, amide, maleimide-thioether, disulfide, phosphodiester, sulfonamide, a product of a clinching reaction or carbamate linkage.

16. Double-stranded iRNA agent according to any one of claims 1 to 15, characterized in that said iRNA agent comprises a single-stranded overhang at at least one of the terminals.

17. Double-stranded RNA agent according to claim 16, characterized in that said single-stranded overhang is 1, 2 or 3 nucleotides in length.

18. Double-stranded RNA agent according to any one of claims 1 to 17, characterized in that the lipophilic portion is conjugated to a nucleobase, sugar portion or internucleoside bond.

19. Double-stranded RNA agent according to any one of claims 1 to 18, characterized in that it further comprises a targeting ligand that targets a receptor that mediates delivery to a CNS tissue.

20. Double-stranded RNA agent according to claim 19, characterized in that the targeting ligand is selected from the group consisting of Angiopep-2, lipoprotein receptor-related protein (LRP) ligand, bEnd.3 cell-binding ligand, transferrin receptor (TfR) ligand, mannose receptor ligand, glucose transporter protein and LDL receptor ligand.

21. Double-stranded RNA agent according to any one of claims 1 to 20, characterized in that the lipophilic portion or the targeting ligand is conjugated by means of a biocleavable ligand selected from the group consisting of DNA, RNA, disulfide, amide, monosaccharides or oligosaccharides functionalized with galactosamine, glucosamine, glucose, galactose, mannose and combinations thereof.

22. Double-stranded RNA agent according to any one of claims 1 to 20, characterized in that the 3' end of the sense strand is protected by a terminal cap which is a cyclic group with an amine, said cyclic group being selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuranyl and decalinyl.

23. Use of a double-stranded RNA agent as defined in any one of claims 1 to 22, characterized in that it is used in the manufacture of a pharmaceutical composition to reduce the expression of a target gene in a cell or individual in sufficient quantity to inhibit the expression of the target gene.

24. Use of a double-stranded RNA agent as defined in any one of claims 1 to 22, characterized in that it is in the manufacture of a pharmaceutical composition or medicament for the treatment of an individual with a CNS disorder or eye disorder.

25. Use according to claim 23 or 24, characterized in that the double-stranded RNA agent is adapted to come into contact with an extrahepatic cell or is adapted to be used by the subject via extrahepatic, intrathecal or intravitreal routes.

26. Use according to claim 23 or 24, characterized in that the pharmaceutical composition is adapted to reduce the expression of a target gene in brain or spinal cord tissue.

27. Use according to claim 26, characterized in that the brain or spinal cord tissue is selected from the group consisting of the cortex, cerebellum, cervical spine, lumbar spine, and thoracic spine.

28. Use according to claim 26, characterized in that the target gene is selected from the group consisting of APP, ATXN2, C9orf72, TARDBP, MAPT(Tau), HTT, SNCA, FUS, ATXN3, ATXN1, SCA1, SCA7, SCA8, MeCP2, PRNP, SOD1, DMPK and TTR.

29. Use according to claim 23 or 24, characterized in that the pharmaceutical composition is adapted to reduce the expression of a target gene in an ocular tissue.

30. Use according to claim 24, characterized in that the CNS disorder is selected from the group of Alzheimer's disease, amyotrophic lateral sclerosis (ALS), frontotemporal dementia, Huntington's disease, Parkinson's disease, spinocerebellar disease, prion disease and Lafora disease.

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