Novel SCN10A RNAi agents and their use

Abstract

The present invention relates to a therapeutic compound, a novel RNA interference (RNAi) agent, that reduces the expression of the voltage-gated sodium channel α subunit 10 (SCN10A) gene, which encodes Nav1.8 or a subunit of the Nav1.8 sodium channel. Such RNAi agents reduce the level of intact Nav1.8 sodium channels and are useful in the treatment of diseases in which the regulation of SCN10A expression and function is involved, such as chronic pain.

Description

Technical Field

[0001] (Reference to Sequence Listing) This application is filed with a sequence listing in ST.26 XML format. The sequence listing is provided as a file titled "30493_WO.xml" created on May 8, 2024, and has a size of 1.7 megabytes. The sequence expression information in ST.26 XML format is hereby incorporated by reference in its entirety into this specification.

[0002] The present invention relates to a therapeutic compound that reduces the expression of the voltage-dependent sodium channel α subunit 10 (SCN10A) gene encoding Nav1.8 or a subunit of Nav1.8 sodium channel, and a novel RNA interference (RNAi) agent. Such RNAi agents reduce the level of intact Nav1.8 sodium channels and are useful in the treatment of diseases such as chronic pain, where regulation of SCN10A expression and function is involved.

[0003] Pain results from neural pathways in response to tissue damage or potential tissue damage signals, may become neuropathic, and may not occur in response to stimuli. Neuropathic pain tends to be associated with dysfunction of the nervous system and is often chronic.

[0004] Voltage-dependent sodium channels (VGSCs) play an important role in the generation and propagation of action potentials. The rapid influx of sodium ions through VGSCs creates the rising phase of action potentials in sensory nociceptive neurons (Blair and Bean, The Journal of Neuroscience, December 1, 2002, 22(23):10277-10290). The Nav1.8 channel plays a major role, in particular, in regulating the firing properties of DRG neurons.

[0005] Chronic pain is a significant unmet need that affects approximately 25% of the world's population. Current treatments result in resistance and / or non-selective inhibition of multiple VGSCs.

[0006] There remains a need for treatments of pain (including chronic pain) that demonstrate improved pain management compared to one or more pain therapies (including greater reduction in pain levels or longer duration of pain relief), therapies requiring fewer physician visits or lower doses compared to one or more pain therapies, therapies that reduce tolerance, improved toxicity profile, improved safety profile, fewer side effects, and / or improved tolerability. [Overview of the Initiative]

[0007] RNAi agents disclosed herein for the treatment of pain, including but not limited to chronic pain, may exhibit improved targeted knockdown, safety, and / or tolerability. RNAi agents herein may further exhibit further improved properties, or any combination of two or more of the aforementioned properties.

[0008] Accordingly, in some embodiments, disclosed herein is an SCN10A RNAi agent comprising a sense strand and an antisense strand, wherein the antisense strand comprises a sequence complementary to a sequence selected from any one of SEQ ID NOs: 2-32, and the sense strand and antisense strand form a double helix. In some embodiments, the double helix comprises a sense strand and an antisense strand that are complementary over at least 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides. In some embodiments of the double helix, the antisense strand comprises a sequence complementary to at least 15, 16, 17, or 18 consecutive nucleotides from any one of SEQ ID NOs: 2-32. In some embodiments of the double helix, the antisense strand comprises a sequence complementary to 15, 16, 17, or 18 consecutive nucleotides from any one of SEQ ID NOs: 2-32. In some embodiments of the double helix, the sense strand is 21 nucleotides long and contains one of the sequences 33 to 63. In some embodiments of the double helix, the antisense strand is 23 nucleotides long and contains one of the sequences 64 to 93.

[0009] In some embodiments, the RNAi agent comprises a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a double helix. a. A sense strand having at least 90% sequence identity with SEQ ID NO: 33, and an antisense strand having at least 90% sequence identity with SEQ ID NO: 64. b. A sense strand having at least 90% sequence identity with SEQ ID NO: 34, and an antisense strand having at least 90% sequence identity with SEQ ID NO: 65. c. A sense strand having at least 90% sequence identity with SEQ ID NO: 35, and an antisense strand having at least 90% sequence identity with SEQ ID NO: 66. d. A sense strand having at least 90% sequence identity with sequence number 36, and an antisense strand having at least 90% sequence identity with sequence number 67. e. A sense strand having at least 90% sequence identity with sequence number 37, and an antisense strand having at least 90% sequence identity with sequence number 68. f. A sense strand having at least 90% sequence identity with sequence number 38, and an antisense strand having at least 90% sequence identity with sequence number 69. g. A sense strand having at least 90% sequence identity with SEQ ID NO: 39, and an antisense strand having at least 90% sequence identity with SEQ ID NO: 70. h. A sense strand having at least 90% sequence identity with sequence number 40, and an antisense strand having at least 90% sequence identity with sequence number 71, i. A sense strand having at least 90% sequence identity with sequence number 41, and an antisense strand having at least 90% sequence identity with sequence number 72, j. A sense strand having at least 90% sequence identity with sequence number 42, and an antisense strand having at least 90% sequence identity with sequence number 73. k. A sense strand having at least 90% sequence identity with sequence number 43, and an antisense strand having at least 90% sequence identity with sequence number 74. l. A sense strand having at least 90% sequence identity with sequence number 44, and an antisense strand having at least 90% sequence identity with sequence number 75. m. A sense strand having at least 90% sequence identity with sequence number 45, and an antisense strand having at least 90% sequence identity with sequence number 76.

[0010] n. A sense strand having at least 90% sequence identity with sequence number 46, and an antisense strand having at least 90% sequence identity with sequence number 77. o. A sense strand having at least 90% sequence identity with sequence number 47, and an antisense strand having at least 90% sequence identity with sequence number 78. p. A sense strand having at least 90% sequence identity with sequence number 48, and an antisense strand having at least 90% sequence identity with sequence number 79, q. A sense strand having at least 90% sequence identity with sequence number 49, and an antisense strand having at least 90% sequence identity with sequence number 80. r. A sense strand having at least 90% sequence identity with sequence number 50, and an antisense strand having at least 90% sequence identity with sequence number 81. s. A sense strand having at least 90% sequence identity with sequence number 51, and an antisense strand having at least 90% sequence identity with sequence number 82. t. A sense strand having at least 90% sequence identity with sequence number 52, and an antisense strand having at least 90% sequence identity with sequence number 83. u. A sense strand having at least 90% sequence identity with sequence number 53, and an antisense strand having at least 90% sequence identity with sequence number 84. v. A sense strand having at least 90% sequence identity with sequence number 54, and an antisense strand having at least 90% sequence identity with sequence number 85. w. A sense strand having at least 90% sequence identity with sequence number 55, and an antisense strand having at least 90% sequence identity with sequence number 86. x. A sense strand having at least 90% sequence identity with sequence number 56, and an antisense strand having at least 90% sequence identity with sequence number 87. y. A sense strand having at least 90% sequence identity with sequence number 57, and an antisense strand having at least 90% sequence identity with sequence number 88. z. A sense strand having at least 90% sequence identity with sequence number 58, and an antisense strand having at least 90% sequence identity with sequence number 89.

[0011] aa. A sense strand having at least 90% sequence identity with sequence number 59, and an antisense strand having at least 90% sequence identity with sequence number 90. bb. A sense strand having at least 90% sequence identity with sequence number 60, and an antisense strand having at least 90% sequence identity with sequence number 91. cc. A sense strand having at least 90% sequence identity with sequence number 61, and an antisense strand having at least 90% sequence identity with sequence number 92, dd. A sense strand having at least 90% sequence identity with sequence number 62, and an antisense strand having at least 90% sequence identity with sequence number 93, or An RNAi agent formed by a pair of oligonucleotide sequences selected from the group consisting of a sense strand having at least 90% sequence identity with SEQ ID NO: 63, and an antisense strand having at least 90% sequence identity with SEQ ID NO: 66.

[0012] In some embodiments, the RNAi agent comprises a sense strand and an antisense strand pair as described in Tables 2, 3A, and 3B.

[0013] In some embodiments, the RNAi agent includes a delivery portion. In some embodiments, the delivery portion is a lipophilic delivery portion. In some embodiments, the delivery portion is conjugated to a nucleotide at position 12 or 13 from the 5' end of the sense strand. In some embodiments, the lipophilic delivery portion is optionally of formula I or 2'-O-hexadecyl. In some embodiments, the sense strand and the antisense strand each independently contain one or more modified nucleotides, and optionally, one or more nucleotide-to-nucleotide bonds of the sense strand or antisense strand are modified nucleotide-to-nucleotide bonds. In some embodiments, one or more modified nucleotides are independently 2'-fluoro-modified nucleotides or 2'-O-methyl-modified nucleotides. In some embodiments, each nucleotide of the sense strand and each nucleotide of the antisense strand are modified.

[0014] Further embodiments provide the use of the RNAi agent of the present invention in pain therapy, including cases where the therapy is for chronic pain. Embodiments also provide, but are not limited, methods for treating pain, including chronic pain, which include administering a therapeutic dose of the RNAi agent of the present invention or a pharmaceutical composition thereof to a subject. [Modes for carrying out the invention]

[0015] The RNAi agent of the present specification includes a sense strand and an antisense strand, each of which is an oligonucleotide. As used herein, "nucleotide" means an organic compound having a nucleoside (e.g., a nucleobase such as adenine, cytosine, guanine, thymine, or uracil, and a pentose sugar such as ribose or 2'-deoxyribose) and a phosphate group. A "nucleotide" can be a "modified nucleotide". A "nucleotide" can function as a monomer unit of nucleic acid polymers such as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).

[0016] As used herein, "oligonucleotide" means a short nucleic acid compound (e.g., less than about 100 nucleotides in length). An oligonucleotide can be single-stranded (ss) or double-stranded (ds). An oligonucleotide can have a double-stranded region or not. As a non-limiting set of examples, an oligonucleotide can be a small interfering RNA (siRNA), a microRNA (miRNA), a short hairpin RNA (shRNA), a Dicer substrate interfering RNA (DsiRNA), or an antisense oligonucleotide (ASO), but is not limited thereto.

[0017] As used herein, "ribonucleotide" means a nucleotide having ribose as its pentose sugar and containing a hydroxyl group at the 2'-position. A modified ribonucleotide is a ribonucleotide having one or more modifications or substitutions of atoms other than hydrogen at the 2'-position, including modifications or substitutions of the nucleobase, sugar, or phosphate group.

[0018] As used herein, "modified internucleotide linkage" means an internucleotide linkage having one or more chemical modifications when compared to a reference internucleotide linkage having a phosphodiester linkage. A modified internucleotide linkage can be a non-naturally occurring linkage.

[0019] As used herein, "modified nucleotide" refers to a nucleotide having one or more chemical modifications when compared to the corresponding reference nucleotide selected from adenosine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide, adenosine deoxyribonucleotide, guanine deoxyribonucleotide, cytosine deoxyribonucleotide, and thymidine deoxyribonucleotide. A modified nucleotide can be a non-naturally occurring nucleotide. A modified nucleotide can have one or more chemical modifications, for example, to its sugar, nucleobase, and / or phosphate group, or any combination thereof. Additionally or alternatively, a modified nucleotide may have one or more chemical moieties conjugated to the corresponding reference nucleotide.

[0020] As used herein, "phosphate analog" means a chemical moiety that mimics the electrostatic and / or steric properties of a phosphate group. In some embodiments, a phosphate analog is located at the 5'-terminal nucleotide of an oligonucleotide instead of a 5'-phosphate. A 5'-phosphate analog can include a phosphatase-resistant linkage. Examples of phosphate analogs include, but are not limited to, 5'-phosphonates such as 5'-methylenephosphonate (5'-MP) and 5'-(E)-vinylphosphonate (5'-VP). An oligonucleotide can have a phosphate analog (referred to as a "4'-phosphate analog") at the 4'-carbon position of the sugar of the 5'-terminal nucleotide. An example of a 4'-phosphate analog is oxymethylphosphonate in which an oxygen atom of an oxymethyl group is bonded to the sugar moiety (e.g., of its 4'-carbon) or an analog thereof. See, for example, International Publication No. WO 2018 / 045317.

[0021] Other modifications to the 5' end of oligonucleotides have been developed (see, for example, International Publication No. 2011 / 133871, U.S. Patent No. 8,927,513, and Prakash et al. (2015) Nuc. Acids Res. 43:2993-3011).

[0022] The term “sequence identity percentage” with respect to a reference nucleic acid sequence is defined as the percentage of nucleotides, nucleosides, or nucleic acid bases in a candidate sequence that are identical to a nucleotide, nucleoside, or nucleic acid base in the reference nucleic acid sequence, after the sequences have been optimally aligned and gaps or overhangs have been introduced as necessary to achieve the maximum sequence identity percentage. Alignment for the purpose of determining nucleic acid sequence identity percentage can be achieved in various ways within the scope of the skill of the art, for example, using publicly available computer software programs, such as those described in Current Protocols in Molecular Biology (Ausubel et al., ed., 1987, Supp. 30, Section 7.7.18, Table 7.7.1), including BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR), Clustal W2.0, or Clustal X2.0 software. A person skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm required to achieve the maximum alignment over the entire length of the sequences being compared. The "sequence identity" percentage can be determined by comparing two optimally aligned sequences across a comparison window, where the nucleic acid sequence fragments within the comparison window may contain additions or deletions (e.g., gaps or overhangs) compared to a reference sequence (without additions or deletions) for optimal alignment of the two sequences. The percentage can be calculated by determining the number of positions where identical nucleotides, nucleosides, or nucleic acid bases exist in both sequences, finding the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the sequence identity percentage. The output is the identity percentage of the target sequence to the query sequence.In some embodiments, the sequence identity percentage is the percentage of nucleotide residues identical between the two strands, calculated using PID3 calculation, which is obtained by dividing the number of identical nucleotide residues by the total number of nucleotides in the shorter of the two sequences and multiplying by 100. See, for example, Raghava, G., Barton, GJ, Quantification of the variation in percentage identity for protein sequence alignments. BMC Bioinformatics 7, 415 (2006). In one embodiment described herein, the alignment is performed using Clustal W2.0. In another embodiment, the alignment is performed using Clustal X2.0. In any of the embodiments described above, following the Clustal alignment, the percentage of nucleotide residues identical between the two strands is calculated using PID3 calculation.

[0023] As used herein, “complementary” or “complementarity” means a structural relationship between two nucleotides, nucleosides, or nucleic acid bases that enables them to form base pairs with each other (e.g., on two opposing nucleic acids or on opposing regions of a single nucleic acid chain, e.g., in a hairpin). For example, a purine nucleotide of one nucleic acid that is complementary to a pyrimidine nucleotide of an opposing nucleic acid may form base pairs with each other by forming hydrogen bonds. Complementary nucleotides can base pair in the classic Watson-Crick scheme, meaning adenine pairs with thymine or uracil, and guanine pairs with cytosine, or in any other way that enables the formation of a stable double helix. Similarly, two nucleic acids may have regions of multiple nucleotides that are complementary to each other and form a complementary region. As used herein, “complementary region” means a nucleotide sequence of nucleic acid that is complementary to an antiparallel nucleotide sequence in order to enable hybridization between two sequences of nucleotides under appropriate hybridization conditions (e.g., in phosphate buffer, in a cell, etc.). In some embodiments, the antisense oligonucleotide includes a region complementary to the mRNA target sequence.

[0024] As used herein, “double helix” means a structure formed by hydrogen bonding of complementary base pairs of two antiparallel sequences of nucleotides under conditions favorable to promote such a structure. Double helix can be formed even without complete complementarity between the two strands or in the presence of debasalized nucleotides.

[0025] As used herein, “iRNA,” “iRNA agent,” “RNAi,” “RNAi agent,” “siRNA,” and “RNA interferant” mean an agent containing RNA that mediates targeted cleavage of RNA transcripts via RNA interference, for example, through the RNA-induced silencing complex (RISC) pathway. In some embodiments, the sense and antisense strands of the RNAi agent are 21–23 nucleotides long. In other embodiments, the sense and antisense strands may be longer, for example, 25–30 nucleotides long, in which case the longer RNAi sequence is processed first by the Dicer enzyme. iRNAs attenuate, inhibit, regulate, or reduce the expression of a target gene in a cell.

[0026] As used herein, “conjugated to,” “conjugate,” or “conjugated” means either an indirect or direct covalent bond. For example, a sense or antisense strand disclosed herein may be covalently bonded to a delivery portion to facilitate entry into cells of CNS tissue. In some embodiments, the sense or antisense strand is directly covalently bonded to the delivery portion. In other embodiments, the sense or antisense strand is indirectly conjugated via a linker, i.e., the sense or antisense strand is covalently bonded to an atom of the linker, which is then covalently bonded to the delivery portion.

[0027] As used herein, “linker” means a structure used to conjugate a nucleotide (e.g., an oligonucleotide) to a targeting ligand or delivery portion. In some embodiments, the linker may be “unstable” or “cleavable,” meaning a linker that can be cleaved (e.g., by acidic pH or enzymes). In other embodiments, the linker may be “stable” or “incleavable,” meaning a linker that is not cleaved under physiological conditions.

[0028] As used herein, “reduced expression,” “decreased expression,” or “decreased expression,” including those relating to genes, means a decrease in the amount or level of RNA transcripts and / or proteins encoded by a gene, and / or a decrease in the level of gene activity, in a cell, cell population, sample, or subject, compared to a suitable reference (e.g., a reference cell, cell population, sample, or subject). For example, when an RNAi agent as used herein (e.g., an oligonucleotide having an antisense strand with a nucleotide sequence complementary to SCN10A mRNA or a portion thereof) is introduced into cells, the amount or level of mRNA, protein, and / or activity may decrease compared to cells not treated with the RNAi agent. Specifically, as used herein, “decreased SCN10A expression” means a decrease in the amount or level of SCN10A mRNA, SCN10A protein levels, and / or SCN10A protein activity in a cell, cell population, sample, or subject, compared to a suitable reference (e.g., a reference cell, cell population, tissue, or subject) not treated with the RNAi agent of the present invention.

[0029] As used herein, “patient” means any mammal, including cats, dogs, mice, rats, and primates, as well as humans, that expresses or has been engineered to express the human Nav1.8 gene. Furthermore, “patient” and “subject” may be used interchangeably.

[0030] As used herein, “treatment” or “to treat” means any process that can slow, control, delay, or halt the progression of a disorder or disease disclosed herein, and does not necessarily mean the complete disappearance of all symptoms of the disorder or disease. Treatment includes the administration of RNAi agents or their pharmaceutical compositions for treating a disease or condition in a mammal, including humans.

[0031] RNAi agents having a sense strand and an antisense strand engineered to reduce the expression of the SCN10A gene are disclosed herein. One embodiment is an RNAi agent having a sense strand and an antisense strand, wherein the antisense strand is complementary to the mRNA transcript of the SCN10A gene, and the antisense strand and sense strand form a double helix.

[0032] In certain embodiments, RNAi agents can reduce the expression of the SCN10A gene in cells. In other embodiments, RNAi agents can reduce the expression of the SCN10A gene in human DRG cells. In other embodiments, RNAi agents can reduce the expression of the SCN10A gene in vivo.

[0033] As described above, RNAi agents can be modified. Therefore, in further embodiments, the antisense strand or sense strand optionally comprises one or more modified nucleotides, and optionally, one or more nucleotide-to-nucleotide bonds of the sense strand or antisense strand are modified nucleotide-to-nucleotide bonds.

[0034] In one embodiment, the RNAi agent comprises a sense strand and an antisense strand, the antisense strand comprising a sequence complementary to a sequence selected from any one of SEQ ID NOs: 2 to 32, the sense strand and the antisense strand forming a double helix, optionally each comprising independently one or more modified nucleotides, and optionally one or more nucleotide-to-nucleotide bonds of the sense strand or the antisense strand being modified nucleotide bonds.

[0035] In further embodiments, the antisense strand includes 3' overhangs of two nucleotides. In further embodiments, the antisense strand and the sense strand independently consist of 15 to 50 nucleotides. In further embodiments, the antisense strand and the sense strand independently consist of 18 to 23 nucleotides. In further embodiments, the antisense strand is 23 nucleotides long. In further embodiments, the sense strand is 21 nucleotides long. In further embodiments, the antisense strand is 23 nucleotides long and the sense strand is 21 nucleotides long.

[0036] In some embodiments, the RNAi comprises a sense strand and an antisense strand forming a double helix, the antisense strand being 23 nucleotides long and the sense strand being 21 nucleotides long. In some embodiments of the double helix, the antisense strand contains a sequence complementary to at least 15, 16, 17, or 18 consecutive nucleotides of a sequence selected from any one of SEQ ID NOs: 2-32. In some embodiments, the double helix comprises a sense strand and an antisense strand that are complementary over at least 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides, or complementary over 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides. In further embodiments, the antisense strand contains one of the sequences from SEQ ID NOs: 64-93. In further embodiments, the sense strand contains one of the sequences from SEQ ID NOs: 33-63. In some embodiments, the RNAi agent comprises a sense strand and an antisense strand pair as described in Table 2, Table 3A, or Table 3B. In some embodiments of the double helix, the sense strand includes SEQ ID NO: 35 and the antisense strand includes SEQ ID NO: 66. In some embodiments, the sense strand includes SEQ ID NO: 39 and the antisense strand includes SEQ ID NO: 70.

[0037] In some embodiments of the RNAi agent, the sense strand and antisense strand form a double helix. a. The sense strand has at least 90% sequence identity with SEQ ID NO: 33, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 64, b. The sense strand has at least 90% sequence identity with SEQ ID NO: 34, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 65, c. The sense strand has at least 90% sequence identity with SEQ ID NO: 35, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 66, d. The sense strand has at least 90% sequence identity with SEQ ID NO: 36, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 67, e. The sense strand has at least 90% sequence identity with SEQ ID NO: 37, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 68, f. The sense strand has at least 90% sequence identity with SEQ ID NO: 38, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 69, g. The sense strand has at least 90% sequence identity with SEQ ID NO: 39, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 70, h. The sense strand has at least 90% sequence identity with SEQ ID NO: 40, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 71, i. The sense strand has at least 90% sequence identity with SEQ ID NO: 41, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 72, j. The sense strand has at least 90% sequence identity with SEQ ID NO: 42, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 73, k. The sense strand has at least 90% sequence identity with SEQ ID NO: 43, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 74, l. The sense strand has at least 90% sequence identity with SEQ ID NO: 44, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 75, The m. sense strand has at least 90% sequence identity with SEQ ID NO: 45, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 76,

[0038] n. The sense strand has at least 90% sequence identity with SEQ ID NO: 46, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 77, o. The sense strand has at least 90% sequence identity with sequence number 47, and the antisense strand has at least 90% sequence identity with sequence number 78, The p-sense strand has at least 90% sequence identity with SEQ ID NO: 48, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 79, q. Does the sense strand have at least 90% sequence identity with sequence number 49, and does the antisense strand have at least 90% sequence identity with sequence number 80? r. The sense strand has at least 90% sequence identity with sequence number 50, and the antisense strand has at least 90% sequence identity with sequence number 81, s. The sense strand has at least 90% sequence identity with sequence number 51, and the antisense strand has at least 90% sequence identity with sequence number 82, t. The sense strand has at least 90% sequence identity with sequence number 52, and the antisense strand has at least 90% sequence identity with sequence number 83, u. The sense strand has at least 90% sequence identity with sequence number 53, and the antisense strand has at least 90% sequence identity with sequence number 84, v. The sense strand has at least 90% sequence identity with SEQ ID NO: 54, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 85, w. The sense strand has at least 90% sequence identity with SEQ ID NO: 55, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 86, x. The sense strand has at least 90% sequence identity with SEQ ID NO: 56, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 87, y. The sense strand has at least 90% sequence identity with sequence number 57, and the antisense strand has at least 90% sequence identity with sequence number 88, z. The sense strand has at least 90% sequence identity with sequence number 58, and the antisense strand has at least 90% sequence identity with sequence number 89,

[0039] aa. The sense strand has at least 90% sequence identity with SEQ ID NO: 59, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 90, bb. The sense strand has at least 90% sequence identity with sequence number 60, and the antisense strand has at least 90% sequence identity with sequence number 91, cc. The sense strand has at least 90% sequence identity with sequence number 61, and the antisense strand has at least 90% sequence identity with sequence number 92, dd. The sense strand has at least 90% sequence identity with SEQ ID NO: 62, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 93, or ee. The sense strand has at least 90% sequence identity with sequence number 63, and the antisense strand has at least 90% sequence identity with sequence number 66.

[0040] In some embodiments of RNAi agents, the double helix comprises a sense strand and an antisense strand. a. The sense strand contains sequence number 94, and the antisense strand contains sequence number 132. b. The sense strand contains SEQ ID NO: 95, and the antisense strand contains SEQ ID NO: 133. c. The sense strand contains SEQ ID NO: 96, and the antisense strand contains SEQ ID NO: 134. d. The sense strand contains sequence number 97, and the antisense strand contains sequence number 135. e. The sense strand contains SEQ ID NO: 98, and the antisense strand contains SEQ ID NO: 136. f. The sense strand contains sequence number 99 and the antisense strand contains sequence number 137. g. The sense strand contains SEQ ID NO: 100, and the antisense strand contains SEQ ID NO: 138. h. The sense strand contains SEQ ID NO: 101, and the antisense strand contains SEQ ID NO: 139. i. The sense strand contains sequence number 102, and the antisense strand contains sequence number 140. j. The sense strand contains SEQ ID NO: 103 and the antisense strand contains SEQ ID NO: 141. k. The sense strand contains SEQ ID NO: 104, and the antisense strand contains SEQ ID NO: 142. l. The sense strand contains SEQ ID NO: 105, and the antisense strand contains SEQ ID NO: 143. The m. sense strand contains SEQ ID NO: 106, and the antisense strand contains SEQ ID NO: 144.

[0041] n. The sense strand contains sequence number 107 and the antisense strand contains sequence number 145. o. The sense strand contains sequence number 108, and the antisense strand contains sequence number 146. p.Sense strand contains SEQ ID NO: 109, and antisense strand contains SEQ ID NO: 147. q. The sense strand contains sequence number 110, and the antisense strand contains sequence number 148. r. The sense strand contains sequence number 111, and the antisense strand contains sequence number 149. s. The sense strand contains sequence number 112, and the antisense strand contains sequence number 150. t. The sense strand contains sequence number 113, and the antisense strand contains sequence number 151. u. The sense strand contains sequence number 114, and the antisense strand contains sequence number 152. v. The sense strand contains sequence number 115, and the antisense strand contains sequence number 153. w. The sense strand contains sequence number 116, and the antisense strand contains sequence number 154. x. The sense strand contains sequence number 117, and the antisense strand contains sequence number 155. y. The sense strand contains sequence number 118, and the antisense strand contains sequence number 156. z. The sense strand contains sequence number 119, and the antisense strand contains sequence number 157.

[0042] aa. The sense strand contains sequence number 120, and the antisense strand contains sequence number 158. bb. The sense strand contains sequence number 121, and the antisense strand contains sequence number 159. The sense strand contains sequence number 122, and the antisense strand contains sequence number 160. dd.Sense strand contains SEQ ID NO: 123, and antisense strand contains SEQ ID NO: 161, ee. The sense strand contains sequence number 96, and the antisense strand contains sequence number 162. ff. The sense strand contains sequence number 96, and the antisense strand contains sequence number 163. gg. The sense strand contains sequence number 96, and the antisense strand contains sequence number 164. The sense strand contains sequence number 96, and the antisense strand contains sequence number 165. ii. The sense strand contains sequence number 96, and the antisense strand contains sequence number 166, jj. The sense chain contains sequence number 124, and the antisense chain contains sequence number 162. kk.Sense strand contains SEQ ID NO: 124 and antisense strand contains SEQ ID NO: 163, ll. The sense strand contains sequence number 124, and the antisense strand contains sequence number 164. The mm.sense strand contains sequence number 12, and the antisense strand contains sequence number 165.

[0043] nn. The sense strand contains sequence number 124, and the antisense strand contains sequence number 166. oo. The sense strand contains sequence number 124, and the antisense strand contains sequence number 134, pp. The sense strand contains sequence number 125, and the antisense strand contains sequence number 162. qq. The sense strand contains sequence number 125, and the antisense strand contains sequence number 163. rr. The sense strand contains sequence number 125, and the antisense strand contains sequence number 164. The sense strand contains sequence number 125, and the antisense strand contains sequence number 165. The sense strand contains sequence number 125, and the antisense strand contains sequence number 166. uu.Sense strand contains sequence number 125 and antisense strand contains sequence number 134, vv.Sense strand contains sequence number 126 and antisense strand contains sequence number 162, The sense chain contains sequence number 126, and the antisense chain contains sequence number 163. xx. The sense strand contains sequence number 126, and the antisense strand contains sequence number 164. yy.Sense strand contains sequence number 126, and antisense strand contains sequence number 165. zz. The sense strand contains sequence number 126, and the antisense strand contains sequence number 166.

[0044] aaa. The sense strand contains sequence number 126 and the antisense strand contains sequence number 134. bbb. The sense strand contains sequence number 127, and the antisense strand contains sequence number 167. ccc.Sense strand contains SEQ ID NO: 127, and antisense strand contains SEQ ID NO: 168. ddd. The sense strand contains sequence number 127, and the antisense strand contains sequence number 169. eee.Sense strand contains sequence number 128 and antisense strand contains sequence number 170, fff. The sense strand contains sequence number 128 and the antisense strand contains sequence number 171. ggg.Sense strand contains SEQ ID NO: 128 and antisense strand contains SEQ ID NO: 172 hhh. The sense strand contains sequence number 129 and the antisense strand contains sequence number 173. iii. The sense strand contains sequence number 129 and the antisense strand contains sequence number 174, jjj. The sense strand contains sequence number 129 and the antisense strand contains sequence number 175. kkk.Sense strand contains SEQ ID NO: 130, and antisense strand contains SEQ ID NO: 176, III. The sense strand contains SEQ ID NO: 130, and the antisense strand contains SEQ ID NO: 177. The sense strand contains sequence number 130, and the antisense strand contains sequence number 178.

[0045] nnn. The sense strand contains sequence number 131, and the antisense strand contains sequence number 179. ooo. The sense strand contains sequence number 131, and the antisense strand contains sequence number 180, The ppp. sense strand contains sequence number 131, and the antisense strand contains sequence number 181. qqq.Sense chain contains sequence number 232 and antisense chain contains sequence number 233, rrr. The sense strand contains sequence number 234, and the antisense strand contains sequence number 235. sss.Sense chain contains sequence number 236, and antisense chain contains sequence number 233, The sense strand contains sequence number 237, and the antisense strand contains sequence number 233. uuu.The sense strand contains sequence number 238, and the antisense strand contains sequence number 239, vvv.Sense strand contains SEQ ID NO: 240, and antisense strand contains SEQ ID NO: 239, www.sense_strand contains SEQ ID NO: 238 and antisense_strand contains SEQ ID NO: 235, xxx.Sense strand contains sequence number 240, and antisense strand contains sequence number 235. yyy.Sense strand contains sequence number 241 and antisense strand contains sequence number 235, zzz. The sense strand contains sequence number 238, and the antisense strand contains sequence number 242.

[0046] aaaa. The sense strand contains sequence number 232, and the antisense strand contains sequence number 239, bbbb. The sense strand contains sequence number 240, and the antisense strand contains sequence number 242. cccc.Sense strand contains sequence number 94, and antisense strand contains sequence number 182, dddd. The sense strand contains sequence number 95, and the antisense strand contains sequence number 183, eeee. The sense strand contains sequence number 96, and the antisense strand contains sequence number 184, ffff. The sense strand contains sequence number 97, and the antisense strand contains sequence number 185. gggg. The sense strand contains sequence number 98, and the antisense strand contains sequence number 186. hhhh. The sense strand contains sequence number 99 and the antisense strand contains sequence number 187, iiii. The sense strand contains sequence number 100, and the antisense strand contains sequence number 188, jjjj. The sense strand contains sequence number 101 and the antisense strand contains sequence number 189. kkkk.Sense chain contains sequence number 102 and antisense chain contains sequence number 190, llll. The sense strand contains sequence number 103, and the antisense strand contains sequence number 191, mmmm.Sense strand contains SEQ ID NO: 104 and antisense strand contains SEQ ID NO: 192,

[0047] nnnn. The sense strand contains sequence number 105, and the antisense strand contains sequence number 193, oooo. The sense strand contains sequence number 106, and the antisense strand contains sequence number 194, pppp. The sense strand contains sequence number 107, and the antisense strand contains sequence number 195. qqqq.Sense strand contains sequence number 108 and antisense strand contains sequence number 196, rrrr. The sense strand contains sequence number 109 and the antisense strand contains sequence number 197, ssss. The sense strand contains sequence number 110, and the antisense strand contains sequence number 198. tttt. The sense strand contains sequence number 111, and the antisense strand contains sequence number 199, uuuu. The sense strand contains sequence number 112, and the antisense strand contains sequence number 200, vvvv.Sense strand contains sequence number 113 and antisense strand contains sequence number 201, wwww. The sense chain contains sequence number 114 and the antisense chain contains sequence number 202, xxxx.Sense strand contains sequence number 115 and antisense strand contains sequence number 203, yyyy.Sense chain contains sequence number 116 and antisense chain contains sequence number 204, zzzz. The sense strand contains sequence number 117, and the antisense strand contains sequence number 205.

[0048] aaaaa. The sense strand contains sequence number 118 and the antisense strand contains sequence number 206, bbbbb. The sense strand contains sequence number 119 and the antisense strand contains sequence number 207. ccccc. The sense strand contains sequence number 120, and the antisense strand contains sequence number 208. ddddd. The sense strand contains sequence number 121, and the antisense strand contains sequence number 209, eeeee. The sense strand contains sequence number 122, and the antisense strand contains sequence number 210. fffff. The sense strand contains sequence number 123 and the antisense strand contains sequence number 211. ggggg. The sense strand contains sequence number 96 and the antisense strand contains sequence number 212. hhhhh. The sense strand contains sequence number 96 and the antisense strand contains sequence number 213, iiiii. The sense strand contains sequence number 96 and the antisense strand contains sequence number 214, jjjjj. The sense strand contains sequence number 96 and the antisense strand contains sequence number 215. kkkkk. The sense strand contains sequence number 96, and the antisense strand contains sequence number 216. The sense chain contains sequence number 124, and the antisense chain contains sequence number 212. mmmmm.Sense strand contains SEQ ID NO: 124 and antisense strand contains SEQ ID NO: 213,

[0049] nnnnn. The sense strand contains sequence number 124, and the antisense strand contains sequence number 214, ooooo. The sense strand contains sequence number 124, and the antisense strand contains sequence number 215, ppppp.Sense strand contains sequence number 124 and antisense strand contains sequence number 216, qqqqq. The sense strand contains sequence number 124, and the antisense strand contains sequence number 184, rrrrr. The sense strand contains sequence number 125, and the antisense strand contains sequence number 212, sssss. The sense chain contains sequence number 125, and the antisense chain contains sequence number 213. ttttt. The sense strand contains sequence number 125, and the antisense strand contains sequence number 214. uuuuu. The sense strand contains sequence number 125, and the antisense strand contains sequence number 215, vvvvv.Sense strand contains sequence number 125 and antisense strand contains sequence number 216, wwwww.Sense chain contains sequence number 125 and antisense chain contains sequence number 184, xxxxx.Sense chain contains sequence number 126 and antisense chain contains sequence number 212, yyyyy. The sense chain contains sequence number 126, and the antisense chain contains sequence number 213. zzzzz. The sense strand contains sequence number 126 and the antisense strand contains sequence number 214.

[0050] aaaaaa. The sense chain contains sequence number 126 and the antisense chain contains sequence number 215. bbbbbb. The sense strand contains sequence number 126, and the antisense strand contains sequence number 216. cccccc. The sense strand contains sequence number 126, and the antisense strand contains sequence number 184. dddddd. The sense strand contains sequence number 127, and the antisense strand contains sequence number 217, eeeeee. The sense strand contains sequence number 127, and the antisense strand contains sequence number 218, ffffff. The sense strand contains sequence number 127, and the antisense strand contains sequence number 219. gggggg. The sense strand contains sequence number 128, and the antisense strand contains sequence number 220. hhhhhh. The sense strand contains sequence number 128 and the antisense strand contains sequence number 221, iiiiii. The sense strand contains sequence number 128 and the antisense strand contains sequence number 222, jjjjjj. The sense strand contains sequence number 129 and the antisense strand contains sequence number 223. kkkkkk. The sense chain contains sequence number 129 and the antisense chain contains sequence number 224. llllll. The sense strand contains sequence number 129, and the antisense strand contains sequence number 225, mmmmmm. The sense strand contains sequence number 130, and the antisense strand contains sequence number 226.

[0051] nnnnnn. The sense strand contains sequence number 130, and the antisense strand contains sequence number 227, oooooo. The sense strand contains sequence number 130, and the antisense strand contains sequence number 228, pppppp. The sense strand contains sequence number 131, and the antisense strand contains sequence number 229. qqqqqq. The sense strand contains sequence number 131, and the antisense strand contains sequence number 230. rrrrrr. The sense strand contains sequence number 131, and the antisense strand contains sequence number 231, ssssss. The sense chain contains sequence number 232, and the antisense chain contains sequence number 184. tttttt. The sense strand contains sequence number 234, and the antisense strand contains sequence number 215. uuuuuu. The sense strand contains sequence number 236, and the antisense strand contains sequence number 184, vvvvvv. The sense strand contains sequence number 237, and the antisense strand contains sequence number 184, wwwwww. The sense chain contains sequence number 238, and the antisense chain contains sequence number 213, xxxxxx. The sense strand contains sequence number 240, and the antisense strand contains sequence number 213. yyyyyy. The sense strand contains sequence number 238, and the antisense strand contains sequence number 215. zzzzzz. The sense strand contains sequence number 240, and the antisense strand contains sequence number 215,

[0052] aaaaaaa. The sense chain contains sequence number 241 and the antisense chain contains sequence number 215, bbbbbbb. The sense chain contains sequence number 238 and the antisense chain contains sequence number 212. ccccccc. The sense strand contains sequence number 232, and the antisense strand contains sequence number 213. ddddddd. The sense strand contains sequence number 240, and the antisense strand contains sequence number 214.

[0053] In some embodiments of RNAi agents, the double helix comprises a sense strand and an antisense strand. a. The sense chain consists of sequence number 94, and the antisense chain consists of sequence number 132. b. The sense chain consists of sequence number 95, and the antisense chain consists of sequence number 133. c. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 134. d. The sense strand consists of sequence number 97, and the antisense strand consists of sequence number 135. e. The sense strand consists of sequence number 98, and the antisense strand consists of sequence number 136. f. The sense chain consists of sequence number 99, and the antisense chain consists of sequence number 137. g. The sense strand consists of sequence number 100, and the antisense strand consists of sequence number 138. h. The sense strand consists of sequence number 101, and the antisense strand consists of sequence number 139. i. The sense chain consists of sequence number 102, and the antisense chain consists of sequence number 140. j. The sense strand consists of sequence number 103, and the antisense strand consists of sequence number 141. k. The sense strand consists of sequence number 104, and the antisense strand consists of sequence number 142. l. The sense strand consists of sequence number 105, and the antisense strand consists of sequence number 143. The m. sense strand consists of sequence number 106, and the antisense strand consists of sequence number 144.

[0054] n. The sense strand consists of sequence number 107, and the antisense strand consists of sequence number 145. o. The sense strand consists of sequence number 108, and the antisense strand consists of sequence number 146. p. The sense strand consists of sequence number 109, and the antisense strand consists of sequence number 147. q. The sense chain consists of sequence number 110, and the antisense chain consists of sequence number 148. r. The sense strand consists of sequence number 111, and the antisense strand consists of sequence number 149. s. The sense chain consists of sequence number 112, and the antisense chain consists of sequence number 150. t. The sense strand consists of sequence number 113, and the antisense strand consists of sequence number 151. u. The sense chain consists of sequence number 114, and the antisense chain consists of sequence number 152. v. The sense strand consists of sequence number 115, and the antisense strand consists of sequence number 153. w. The sense chain consists of sequence number 116, and the antisense chain consists of sequence number 154. x. The sense chain consists of sequence number 117, and the antisense chain consists of sequence number 155. y. The sense chain consists of sequence number 118, and the antisense chain consists of sequence number 156. z. The sense chain consists of sequence number 119, and the antisense chain consists of sequence number 157.

[0055] aa. The sense chain consists of sequence number 120, and the antisense chain consists of sequence number 158. bb. The sense strand consists of sequence number 121, and the antisense strand consists of sequence number 159. The sense chain consists of sequence number 122, and the antisense chain consists of sequence number 160. The sense strand consists of sequence number 123, and the antisense strand consists of sequence number 161. ee. The sense strand consists of sequence number 96, and the antisense strand consists of sequence number 162. ff. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 163. gg. The sense strand consists of sequence number 96, and the antisense strand consists of sequence number 164. The sense strand consists of sequence number 96, and the antisense strand consists of sequence number 165. ii. The sense strand consists of sequence number 96, and the antisense strand consists of sequence number 166. jj. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 162. kk. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 163. ll. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 164. The mm. sense strand consists of sequence number 124, and the antisense strand consists of sequence number 165.

[0056] nn. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 166. oo. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 134. The sense strand consists of sequence number 125, and the antisense strand consists of sequence number 162. qq. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 163. rr. The sense strand consists of sequence number 125, and the antisense strand consists of sequence number 164. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 165. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 166. uu.The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 134. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 162. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 163. xx. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 164. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 165. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 166.

[0057] aaa. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 134. bbb. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 167. The ccc.sense chain consists of sequence number 127, and the antisense chain consists of sequence number 168. ddd. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 169. eee. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 170. fff. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 171. ggg.Sense chain consists of sequence number 128 and antisense chain consists of sequence number 172. hhh. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 173. iii. The sense chain consists of sequence number 129 and the antisense chain consists of sequence number 174. jjj. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 175. kkk. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 176. III. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 177. The mmm. sense strand consists of sequence number 130, and the antisense strand consists of sequence number 178.

[0058] nnn. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 179. ooo. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 180. The ppp. sense strand consists of sequence number 131, and the antisense strand consists of sequence number 181. qqq. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 233. rrr. The sense chain consists of sequence number 234, and the antisense chain consists of sequence number 235. The sense chain consists of sequence number 236, and the antisense chain consists of sequence number 233. The sense chain consists of sequence number 237, and the antisense chain consists of sequence number 233. uuu. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 239, vvv. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 239. www.sense_chain consists of sequence number 238 and antisense_chain consists of sequence number 235. xxx. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 235. The sense chain consists of sequence number 241, and the antisense chain consists of sequence number 235. zzz. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 242.

[0059] aaaa. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 239. bbbb. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 242. The sense chain consists of sequence number 94, and the antisense chain consists of sequence number 182. dddd. The sense strand consists of sequence number 95, and the antisense strand consists of sequence number 183. eeee. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 184, ffff. The sense chain consists of sequence number 97, and the antisense chain consists of sequence number 185. gggg. The sense chain consists of sequence number 98, and the antisense chain consists of sequence number 186. hhhh. The sense chain consists of sequence number 99, and the antisense chain consists of sequence number 187. iiii. The sense chain consists of sequence number 100, and the antisense chain consists of sequence number 188. jjjj. The sense chain consists of sequence number 101, and the antisense chain consists of sequence number 189. kkkk. The sense chain consists of sequence number 102, and the antisense chain consists of sequence number 190. The sense chain consists of sequence number 103, and the antisense chain consists of sequence number 191. mmmm. The sense strand consists of sequence number 104, and the antisense strand consists of sequence number 192.

[0060] nnnn. The sense chain consists of sequence number 105, and the antisense chain consists of sequence number 193, oooo. The sense chain consists of sequence number 106, and the antisense chain consists of sequence number 194, pppp. The sense strand consists of sequence number 107, and the antisense strand consists of sequence number 195. qqqq. The sense chain consists of sequence number 108, and the antisense chain consists of sequence number 196. rrrr. The sense strand consists of sequence number 109, and the antisense strand consists of sequence number 197, ssss. The sense chain consists of sequence number 110, and the antisense chain consists of sequence number 198. The sense chain consists of sequence number 111, and the antisense chain consists of sequence number 199. uuuu. The sense chain consists of sequence number 112, and the antisense chain consists of sequence number 200, vvvv. The sense chain consists of sequence number 113, and the antisense chain consists of sequence number 201. wwww. The sense chain consists of sequence number 114, and the antisense chain consists of sequence number 202. xxxx. The sense chain consists of sequence number 115, and the antisense chain consists of sequence number 203. yyyy. The sense chain consists of sequence number 116, and the antisense chain consists of sequence number 204. zzzz. The sense chain consists of sequence number 117, and the antisense chain consists of sequence number 205.

[0061] aaaaa. The sense chain consists of sequence number 118, and the antisense chain consists of sequence number 206. bbbbb. The sense chain consists of sequence number 119 and the antisense chain consists of sequence number 207. The sense chain consists of sequence number 120, and the antisense chain consists of sequence number 208. ddddd. The sense strand consists of sequence number 121, and the antisense strand consists of sequence number 209, eeeee. The sense chain consists of sequence number 122, and the antisense chain consists of sequence number 210. fffff. The sense chain consists of sequence number 123, and the antisense chain consists of sequence number 211. ggggg. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 212. hhhhh. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 213. iiiii. The sense chain consists of sequence number 96 and the antisense chain consists of sequence number 214, jjjjj. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 215. kkkkk. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 216. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 212. mmmmm. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 213.

[0062] nnnnn. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 214, ooooo. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 215, ppppp. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 216. qqqqq. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 184. rrrrr. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 212, sssss. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 213. ttttt. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 214. uuuuu. The sense strand contains sequence number 125, and the antisense strand contains sequence number 215, vvvvv. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 216. wwwww. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 184. xxxxx. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 212. yyyyy. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 213. zzzzz. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 214.

[0063] aaaaaa. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 215. bbbbbb. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 216. cccccc. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 184. dddddd. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 217, eeeeee. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 218, ffffff. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 219. gggggg. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 220. hhhhhh. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 221. iiiiii. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 222, jjjjjj. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 223. kkkkkk. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 224. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 225. mmmmmm. The sense strand consists of sequence number 130, and the antisense strand consists of sequence number 226.

[0064] nnnnnn. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 227, oooooo. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 228, pppppp. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 229. qqqqqq. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 230. rrrrrr. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 231, ssssss. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 184. tttttt. The sense chain consists of sequence number 234, and the antisense chain consists of sequence number 215. uuuuuu. The sense chain consists of sequence number 236, and the antisense chain consists of sequence number 184, vvvvvv. The sense chain consists of sequence number 237, and the antisense chain consists of sequence number 184. wwwwww. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 213. xxxxxx. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 213. yyyyyy. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 215. zzzzzz. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 215.

[0065] aaaaaaa. The sense chain consists of sequence number 241, and the antisense chain consists of sequence number 215. bbbbbbb. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 212. ccccccc. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 213. ddddddd. The sense strand consists of sequence number 240, and the antisense strand consists of sequence number 214.

[0066] In some embodiments of the RNAi agent, the antisense strand is essentially derived from Sequence ID No. 233, and the sense strand is essentially derived from Sequence ID No. 232.

[0067] In some embodiments of the RNAi agent, the antisense strand is essentially derived from Sequence ID No. 235, and the sense strand is essentially derived from Sequence ID No. 234.

[0068] In some embodiments of the RNAi agent, the antisense strand is essentially derived from SEQ ID NO: 233, and the sense strand is essentially derived from SEQ ID NO: 236.

[0069] In some embodiments, the RNAi agent includes a sense strand further comprising a delivery moiety. In some embodiments, the RNAi agent includes an antisense strand further comprising a delivery moiety.

[0070] In some embodiments of RNAi agents, the delivery portion is a lipophilic delivery portion. In one embodiment, the delivery portion is conjugated to a nucleic acid base in the sense strand, as represented by formula Ib. In another embodiment, the delivery portion is conjugated to a nucleic acid base in the sense strand, as represented by the formula in Table 10. In some embodiments, the delivery portion is conjugated to a nucleic acid base at position 12 or 13 from the 5' end of the sense strand.

[0071] In some embodiments, the RNAi agent includes nucleotide modifications. Therefore, in certain embodiments, the RNAi agent includes modified nucleotides that are independently 2'-fluoromodified nucleotides or 2'-O-methyl-modified nucleotides. In some embodiments, each nucleotide in the sense strand and each nucleotide in the antisense strand are modified.

[0072] In certain embodiments, the 2' fluoromodified nucleotide is located at a specific position selected from the following: a. Positions 2, 3, 7, 14, and 16 from the 5' end of the antisense chain, b. Positions 2, 5, 7, 14, and 16 from the 5' end of the antisense chain, c. Positions 2, 3, 8, 14, and 16 from the 5' end of the antisense chain, d. Positions 2, 5, 8, 14, and 16 from the 5' end of the antisense chain, e. Positions 2, 14, and 16 from the 5' end of the antisense chain, or f. Positions 2, 6, 14, and 16 from the 5' end of the antisense chain.

[0073] In certain embodiments, the 2' fluoromodified nucleotide is located at a specific position selected from the following: a. Positions 9, 10, and 11 from the 5' end of the sense strand, b. Positions 7, 9, and 11 from the 5' end of the sense strand, c. Positions 7, 9, and 10 from the 5' end of the sense strand, d. Positions 7, 10, and 11 from the 5' end of the sense strand, or e. Positions 7, 9, 10, and 11 from the 5' end of the sense strand.

[0074] In further embodiments, the 2'-fluoromodified nucleotide is located at a specific position on the antisense strand, and that position is a. Positions 2, 3, 7, 14, and 16 from the 5' end of the antisense chain, b. Positions 2, 5, 7, 14, and 16 from the 5' end of the antisense chain, c. Positions 2, 3, 8, 14, and 16 from the 5' end of the antisense chain, d. Positions 2, 5, 8, 14, and 16 from the 5' end of the antisense chain, e. Positions 2, 14, and 16 from the 5' end of the antisense chain, or f. Positions 2, 6, 14, and 16 from the 5' end of the antisense chain, Selected from, the 2' fluoro-modified nucleotide is located at a specific position on the sense strand, and that position is a. Positions 9, 10, and 11 from the 5' end of the sense strand, b. Positions 7, 9, and 11 from the 5' end of the sense strand, c. Positions 7, 9, and 10 from the 5' end of the sense strand, d. Positions 7, 10, and 11 from the 5' end of the sense strand, or e. From the 5' end of the sense strand, positions 7, 9, 10 and 11, and f. Positions 7, 9, 10, and 11 from the 5' end of the sense strand, and positions 2, 6, 14, and 16 from the 5' end of the antisense strand. g. Selected from positions 9, 10, and 11 from the 5' end of the sense strand, and positions 2, 5, 8, 14, and 16 from the 5' end of the antisense strand.

[0075] In some embodiments of the RNAi agent, the nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand, and at positions 7, 9, 10, and 11 from the 5' end of the sense strand, include 2' fluoromodifications.

[0076] In some embodiments of the RNAi agent, the nucleotides at positions 2, 5, 7, 14, and 16 from the 5' end of the antisense strand, and at positions 9, 10, and 11 from the 5' end of the sense strand, contain 2' fluoromodifications.

[0077] In some embodiments of the RNAi agent, the nucleotides at positions 2, 3, 7, 14, and 16 from the 5' end of the antisense strand, and positions 9, 10, and 11 from the 5' end of the sense strand, contain 2' fluoromodifications.

[0078] In some embodiments of the RNAi agent, the nucleotides at positions 2, 5, 8, 14, and 16 from the 5' end of the antisense strand, and at positions 9, 10, and 11 from the 5' end of the sense strand, include 2' fluoromodifications.

[0079] In some embodiments of the RNAi agent, the nucleotides at positions 2, 14, and 16 from the 5' end of the antisense strand, and at positions 9, 10, and 11 from the 5' end of the sense strand, include 2' fluoromodifications.

[0080] In some embodiments of the RNAi agent, the nucleotides at positions 2, 5, 7, 14, and 16 from the 5' end of the antisense strand, and at positions 7, 9, 10, and 11 from the 5' end of the sense strand, include 2' fluoromodifications.

[0081] In some embodiments of RNAi agents, in which a nucleotide at a specific position is modified to include a 2'-fluoromodified nucleotide, the remaining nucleotides are also modified, optionally including a 2'-O-methylmodified nucleotide.

[0082] In some embodiments, the RNAi agent includes one or more modified nucleoside links. In some embodiments, the nucleoside links are phosphorothioate chains. In some embodiments, the antisense strand has four or five phosphorothioate chains. In some embodiments, the sense strand has four or five phosphorothioate chains. In some embodiments, the antisense strand includes four or fewer phosphorothioate nucleotide links. In some embodiments, positions 1 and 2 of the sense strand (e.g., relative to the 5' or 3' end) are linked by modified nucleotide links. In some embodiments, positions 1 and 2 of the antisense strand (e.g., relative to the 5' or 3' end) are linked by phosphorothioate nucleotide links. In some embodiments, positions 2 and 3 of the sense strand (e.g., relative to the 5' or 3' end) are linked by modified nucleotide links. In some embodiments, positions 2 and 3 of the antisense strand (e.g., relative to the 5' or 3' end) are linked by phosphorothioate nucleotide links.

[0083] In certain embodiments, the 5' end of the antisense chain is modified to include a phosphate group or a phosphate analog. In some embodiments, the phosphate analog is a 5'-(E)-vinylphosphonate.

[0084] In some embodiments, the RNAi agent includes a modified nucleotide that is a debasic portion. In some embodiments, the sense strand has a modified nucleotide that is a debasic portion. In some embodiments, the sense strand and antisense strand of the RNAi agent herein form a complementary region of at least 15 nucleotides. In other embodiments, the sense strand and antisense strand form a complementary region of at least 16, 17, or 18 nucleotides. In other embodiments, the sense strand and antisense strand form a complementary region of 15, 16, 17, 18, 19, 20, or 21 nucleotides.

[0085] Another embodiment is an RNAi agent having a sense strand and an antisense strand, wherein the antisense strand is complementary to a portion of the mRNA of Sequence ID No. 1 (transcript of the SCN10A gene), the sense strand and antisense strand form a double helix, each independently being 15 to 25 nucleotides long, and optionally, each independently containing one or more modified nucleotides. In some embodiments, the sense strand and / or antisense strand are selected from the table disclosed herein. In some embodiments, the RNAi agent includes an antisense strand and a sense strand forming a double helix selected from the table disclosed herein.

[0086] Another embodiment disclosed herein is an RNAi agent having formula I:RLD, where R is a double-stranded oligonucleotide having a sense strand and an antisense strand, the sense strand and antisense strand forming a complementary region, D is a delivery portion, L is absent or a linker portion, the antisense strand comprises at least 15 consecutive nucleotides complementary to the mRNA transcript of the SCN10A gene (e.g., SEQ ID NO: 1), the sense strand and antisense strand are complementary over at least 15 nucleotides, in some embodiments the sense strand and antisense strand form a complementary region of 15, 16, 17, or 18 nucleotides, the sense strand and antisense strand are each independently 18 to 23 nucleotides long, optionally the sense strand and antisense strand each independently comprises one or more modified nucleotides, optionally D is present.

[0087] In another embodiment, disclosed herein is an RNAi agent having formula I:RLD, where R is a double-stranded oligonucleotide having a sense strand and an antisense strand, the sense strand and antisense strand forming a double helix, D is a delivery portion, L is absent or a linker portion, the antisense strand comprises at least 15 consecutive nucleotides complementary to the mRNA transcript of the SCN10A gene, the sense strand and antisense strand are each independently 18 to 23 nucleotides long, and optionally, the sense strand and antisense strand each independently comprises one or more modified nucleotides. In a further embodiment, the antisense strand comprises at least 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides complementary to the SCN10A gene.

[0088] Furthermore, disclosed herein are RNAi agents having formula I:RLD, where R is a double-stranded oligonucleotide having a sense strand and an antisense strand, the sense strand and antisense strand forming a double helix, D is a delivery portion, L is absent or a linker portion, the antisense strand is complementary to at least 18 nucleotides of the sequence in Table 1, the sense strand and antisense strand are complementary over at least 15, 16, 17, 18, 19, 20 or 21 nucleotides, the sense strand and antisense strand are independently 18 to 23 nucleotides long, and optionally, the sense strand and antisense strand each independently contain one or more modified nucleotides.

[0089] In some embodiments, the delivery portion is formula I:

[0090] [ka] This is the delivery portion.

[0091] In some embodiments, the RNAi agent comprises a sense strand and an antisense strand, the sense strand and the antisense strand forming a double-stranded region, and the sense strand or antisense strand has the following formula:

[0092] [ka] Includes modified nucleotides, In some embodiments, B is a nucleic acid base selected from adenine (A), cytosine (C), guanine (G), thymine (T), uracil (U), or derivatives thereof. In some embodiments, B is a nucleic acid base selected from A, C, G, T, and U. In some embodiments, B is a nucleic acid base derivative selected from 5-methylcytosine, 2-thiouridine, 4-thiouridine, C5-modified pyrimidine, C2-modified purine, N8-modified purine, pseudouracil, isocytosine, isoguanine, 2,6-dianinopurine, pseudocytosine, 2-aminopurine, xanthine, hypoxanthine, 7-methylguanine, 5-hydroxymethylcytosine, 5,6-dihydrouracil, 5-carboxycytidine, phenoxazine, N6-alkyl-A, or O6-alkyl-G.

[0093] In some embodiments, the sense strand contains a modified nucleotide of formula Ia. In some embodiments, the sense strand contains, for example, one of the modified nucleotides of formula Ia at any one of the positions 1-6 or 12-21 from the 5' end. In some embodiments, the sense strand contains a modified nucleotide of formula Ia at position 13 from the 5' end. In some embodiments, the sense strand contains a modified nucleotide of formula Ia at position 12 from the 5' end.

[0094] In some embodiments, the delivery portion is conjugated to a nucleotide of the sense strand. In some embodiments, the delivery portion is a modified nucleotide located on the sense strand. In some embodiments, the modified nucleotide is located at positions 5' to 12 or 13 of the sense strand. In some embodiments, the modified nucleotide is 2'-O-hexadecyluridine, 2'-O-hexadecylcytidine, 2'-O-hexadecylguanine, or 2'-O-hexadecyladenosine, depending on the sequence of the sense strand at position 12 or 13 (see Table 10 for structures).

[0095] For RNAi agents described herein that reduce the expression of the SCN10A gene, the antisense strand is complementary to the sequence in Table 1. In some embodiments, the antisense strand is complementary to at least 15 nucleotides of the sequence in Table 1. In some embodiments, the antisense strand is complementary to 18 consecutive nucleotides of the sequence in Table 1. In some embodiments, the RNAi agent contains 0, 1, 2, or 3 mismatches between the sense strand and the antisense strand.

[0096] In some embodiments, the antisense or sense strand of the RNAi agent has a sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the antisense or sense strands of Table 2, Table 3A, or Table 3B. In some embodiments, the antisense or sense strand of the RNAi agent has a sequence with 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the antisense or sense strands described in Table 2, Table 3A, or Table 3B.

[0097] In some embodiments, the sense strand and antisense strand are complementary over 15, 16, 17, 18, 19, or 20 nucleotides. In some embodiments, the sense strand and antisense strand are complementary over 15, 16, 17, 18, 19, or 20 consecutive nucleotides. In further embodiments, the RNAi agent contains 0, 1, 2, or 3 mismatches between the sense strand and the antisense strand.

[0098] In some embodiments of the RNAi agents disclosed herein, the RNAi agents can reduce the expression of the SCN10A gene in cells. In some embodiments, the RNAi agents disclosed herein are for therapeutic use. In one embodiment, the use is in the treatment of chronic pain. In some embodiments, the RNAi agents are for use in the manufacture of pharmaceuticals for the treatment of pain, including chronic pain.

[0099] In some embodiments, the RNAi agent is incorporated into a pharmaceutical composition comprising the RNAi agent and one or more pharmaceutically acceptable excipients.

[0100] Another embodiment provided herein is a method for reducing SCN10A protein and / or mRNA expression in cells, comprising contacting the cells with an RNAi agent disclosed herein and incubating the cells for a time sufficient to degrade the SCN10A mRNA. The reduction in SCN10A protein and / or mRNA expression is measured by any suitable assay. In some embodiments, the degradation of SCN10A mRNA is determined by RT-PCR.

[0101] Some embodiments provided herein are methods for treating patients who would benefit from reducing the expression of mRNA transcripts of the SCN10A gene, comprising administering an effective amount of an RNAi agent or a pharmaceutical composition thereof to the patient in need. Some embodiments are methods for treating chronic pain in patients in need of treatment for chronic pain, comprising administering a therapeutic amount of an RNAi agent or a pharmaceutical composition thereof disclosed herein.

[0102] RNAi agents and compositions containing RNAi agents are administered by any preferred method, including, but not limited to, intrathecal delivery (e.g., intrathecal injection (lumbar puncture)), epidural delivery (e.g., transarctic (midline) epidural injection, transforaminal (peri-DRG) epidural injection), systemic delivery such as intravenous or subcutaneous delivery (e.g., intravenous or subcutaneous injection), or intraganglionic injection.

[0103] The drug regimen may be adjusted to provide the optimal desired response (e.g., therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the urgency of the therapeutic situation.

[0104] The dosage values ​​may vary depending on the type and severity of the condition to be alleviated. It should be further understood that for any particular subject, the specific medication regimen should be adjusted over time according to the individual needs and the professional judgment of the person managing or supervising the administration of the composition.

[0105] In another embodiment, the foregoing provides compounds, RNAi agents, or pharmaceutical compositions for use in therapy. The foregoing also provides compounds, RNAi agents, or pharmaceutical compositions for use in the treatment of pain disorders.

[0106] NCBI reference sequence: NM_006514.4 (sequence number 1): Sequence ID 1:

[0107] Table 1

[0108] Table 2

[0109] Table 3

[0110] Table 4

[0111] Table 5

[0112] Table 6

[0113] Table 7-1

[0114] Table 7-2

[0115] Table 8-1

[0116] Table 8-2

[0117] Table 8-3

[0118] Table 8-4

[0119] Table 8-5

[0120] Table 8-6

[0121] Table 9-1

[0122] Table 9-2

[0123] Table 9-3

[0124] Table 9-4

[0125] Table 9-5

[0126] Table 9-6

[0127] In Tables 3A and 3B, modifications are indicated immediately before the corresponding modified nucleotide. The abbreviation "P" represents a phosphate group, "m" represents methylation of an OH group that produces a 2' methoxy modification on ribose, and "f" represents a 2' fluoro modification of ribose. * " represents a phosphorothioate modification of the phosphodiester bond in the skeleton), "ads" represents formula Ia, and Ahd or Uhd represents a delivery moiety that is a 2'-O-hexadecyl nucleic acid base as shown in Table 10. In some embodiments, the RNAi agent includes a "phosphate analog" modification located at the 5' terminal nucleotide of the sense or antisense oligonucleotide instead of a 5'-phosphate. In some embodiments, the 5' modification is a phosphate group ("p"). In some embodiments, the modification is a 5'-(E)-vinylphosphonate (5'-VP or VP). In some embodiments, the sense strand includes a delivery moiety conjugated to the nucleotide at position 12 or 13 from the 5' end of the sense strand, and in some embodiments, the lipophilic delivery moiety is optionally of formula I or 2'-O-hexadecyl. [Examples]

[0128] Example 1: Design of RNAi agent The RNAi agent was designed to contain an antisense strand complementary to the 18-mer region of SCN10A having the sequence of transcript NM_000718.4 (SEQ ID NO: 1), as shown in Table 1. The sense RNA oligonucleotide strand and antisense RNA oligonucleotide strand are 18–23 nucleotides long and have an optional overhang of 1, 2, 3, 4, or 5 nucleotides. These nucleotides are characterized by 1–10 fluoroadditions at the 2' position of ribose, and the remaining nucleotides are methylated at the 2' position of ribose (producing 2'-methoxy, i.e., 2'O-methyl modification).

[0129] In some embodiments, the sense strand and / or antisense strand independently contain 1, 2, 3, 4, or 5 fluoroadditions at the 2' position of ribose, and the remaining nucleotides are methylated at the 2' position of ribose. In some embodiments, the antisense strand is optionally phosphorylated at its 5' end. In other embodiments, the antisense strand is optionally modified with a vinylphosphonic acid group at its 5' end. One or more phosphodiester bonds are present at the 5' and 3' ends of each strand. Optionally, a VPN is present at the 5' end of the antisense strand, for example, instead of a phosphate group.

[0130] Table 1 shows 18-mer sequences used to generate complementary antisense strands. In some embodiments, the antisense strands are designed to be 23 nucleotides long.

[0131] Using the 18mers in Table 1, we generated the double hemispheres shown in Table 2.

[0132] Example 2: In vitro knockdown of SCN10A gene expression The knockdown of SCN10A expression using RNAi agents was assayed using the following procedure.

[0133] Human iPSC-derived sensory neurons (purchased from Anatomic) were plated, and each RNAi agent was added directly to individual wells. RNAi agents derived from the sequences in Table 2 were added in two strips at a single concentration of 1 μM (1000 nM). The treated cells were lysed, and changes in gene expression in RNAi-treated cells were analyzed using Cells-to-CT Kits according to the manufacturer's protocol (ThermoFisher A35377). Pre-designed gene expression assays (supplied as 20× mixtures) were selected from Applied Bio-systems (Foster City, CA, USA). The efficiency of these assays (ThermoFisher Hs_01045139_m1 SCN10A and ThermoFisher Hs99999905_m1 GAPDH) was characterized using cDNA dilution series. RT-QPCR was performed using a MicroAmp Optical 384-well reaction plate with the QuantStudio 7 Flex system. Relative gene expression levels were determined using the delta-delta CT method, which normalizes for the housekeeping gene GAPDH. IC50 was determined using 3-parameter logistic fitting with GraphPad Prism v9.0.

[0134] Selective siRNA agents containing nucleotide sequences were tested at multiple concentrations. Teg-cholesterol conjugate siRNAs were used at concentrations of 4000, 1000, 250, 62.5, 15.6, 3.9, 0.98, and 0.24 nM.

[0135] SCN10A oligonucleotides were conjugated to the lipophilic delivery region as follows. Double helixes with the numbers shown in Tables 4 and 5 were conjugated with teg-cholesterol. The teg-cholesterol conjugates were evaluated for knockdown in iPSC cells in vitro. The sense and antisense strands of the conjugated RNAi agents are shown in Table 3A.

[0136] The results are shown in Table 4 as the knockdown percentage (%KD) of the SCN10A transcript.

[0137] The RNAi agent conjugated as described above is tested in vitro in iPSC cells using the same method. The results of knockdown at a single concentration or various concentrations are shown in Table 4 and Table 5, respectively. Table 5 includes data from multiple concentrations to establish the IC50, but only shows 4 μM and the IC50. The ND entries correspond to cases where the measured values were not determined.

[0138]

Table 10

[0139]

Table 11-1

[0140]

Table 11-2

[0141] Example 3: Knockdown of SCN10A in Cultured Human DRG Cells Selected RNAi agents were further tested in human DRG cultures. DRGs were isolated from deceased human donors and plated in a medium containing DMEM / F12 supplemented with 2% horse serum (ThermoFisher Scientific), 30 ng / mL human NGF-B (N-245; Alomone), 30 ng / mL human GDNF (G-240; Alomone), 1 ng / mL human NT3 (N-260; Alomone), Gem21 NeuroPlex (GeminiBio), and penicillin / streptomycin (1037,8016; ThermoFisher Scientific), and then maintained for 3 days. Then, siRNA (conjugated with Teg-cholesterol) was directly added to the DRG neuronal cell cultures at a final concentration of 1000 nM, and after waiting for 5 days, they were harvested.

[0142] RNA was recovered. Specifically, siRNA-treated cells were lysed, and cellular RNA was isolated using the RNAdvance v2 for Cell kit (A47943; Beckman Coulter). From 60 μL of eluted RNA, 1.5 μL of total RNA was used for RT-qPCR using the Luna Universal Probe One-Step RT-QPCR kit (E3006E; NEB) according to the manufacturer's instructions. Gene expression was measured using the QuanStudio 6 Flex system with Taqman assays for SCN10A (Hs01045137_m1, Thermo Fisher), GAPDH (Hs99999905_m1, Thermo Fisher), and STMN2 (Hs00975900_m1, Thermo Fisher). The delta-delta Ct of the RT-qPCR data was analyzed using Design & Analysis Software 2.6.0 and subsequent data visualization with GraphPad 9.0.

[0143] The results are shown in Table 6 as knockdown percentage (%KD) at the SCN10A transcript level.

[0144] [Table 12]

[0145] Example 4: RNAi agents administered in vivo to rats Lipid-conjugated siRNAs were prepared as described in the Art. A 16-carbon carboxylic acid was conjugated to the nucleotide at position 13 of the sense strand. The 5' end of the antisense strand was modified with a 5'VP moiety. In Table 7, double strand 100 is double strand 32 containing a 16-carbon chain presented detached from the 2' hydroxyl group of the nucleotide at position 13 of the sense strand, and the 5' end of the antisense strand was modified with a 5'VP moiety instead of a 5'P moiety. In Table 7, double strand 101 is double strand 33 containing a 16-carbon chain presented detached from the 2' hydroxyl group of the nucleotide at position 13 of the sense strand, and the 5' end of the antisense strand was modified with a 5'VP moiety instead of a 5'P moiety. In Table 7, double strand 102 is double strand 34 containing a 16-carbon chain presented detached from the 2' hydroxyl group of the nucleotide at position 13 of the sense strand, and the 5' end of the antisense strand was modified with a 5'VP moiety instead of a 5'P moiety. In Table 7, double strand 105 is double strand 37, which includes a 16-carbon chain presented detached from the 2' hydroxyl group of the nucleotide at position 13 of the sense strand, and the 5' end of the antisense strand is modified with a 5'VP moiety instead of a 5'P moiety. In Table 7, double strand 106 is double strand 38, which includes a 16-carbon chain presented detached from the 2' hydroxyl group of the nucleotide at position 13 of the sense strand, and the 5' end of the antisense strand is modified with a 5'VP moiety instead of a 5'P moiety. In Table 7, double strand 107 is double strand 39, which includes a 16-carbon chain presented detached from the 2' hydroxyl group of the nucleotide at position 13 of the sense strand, and the 5' end of the antisense strand is modified with a 5'VP moiety instead of a 5'P moiety.

[0146] Rats expressing the human SCN10A gene at the rat locus were intrathecally administered 4 μg of siRNA or vehicle control. The animals were sacrificed after two weeks, and the cervical dorsal root ganglia (CDRG), lumbar dorsal root ganglia (LDRG), and upper thoracic dorsal root ganglia (TDRG) were isolated. RNA was prepared as described above, and RT-PCR was performed as described above. The percentage of remaining SCN10A mRNA was measured relative to GAPDH. The results are shown in Table 7.

[0147]

Table 13

[0148] Rats expressing the human SCN10A gene at the rat locus were administered 300 μg of siRNA or vehicle control intrathecally. Animals were sacrificed after 1, 2, or 4 weeks (see Table 8), and cervical dorsal root ganglia (CDRG), lumbar dorsal root ganglia (LDRG), and thoracic dorsal root ganglia (TDRG) were isolated. RNA was prepared as described above, and RT-PCR was performed as described above. The percentage of remaining SCN10A mRNA was measured relative to GAPDH. The results are shown in Table 8.

[0149]

Table 14

[0150] Example 5: RNAi Agents Administered In Vivo in Non-Human Primate (NHP) Studies An in vivo test of duplex number 168 was performed in cynomolgus monkeys (Macaca fascicularis) to evaluate the efficacy of SCN10A siRNA. To elucidate the efficacy of siRNA in silencing the target gene, indwelling catheters were implanted intrathecally in the lumbar region in cynomolgus monkeys at n = 4 / group. The monkeys were injected with either aCSF or duplex number 168 (8 mg / mL in aCSF) over 15 minutes and perfusion-fixed 29 days later. Tissues collected at necropsy included dorsal root ganglia, spinal cord, nerves, and skin biopsies. qPCR was performed to determine mRNA knockdown in the target region of interest. The following Table 9 shows the remaining SCN10A mRNA observed in the lumbar DRG 29 days after a single administration of siRNA.

[0151]

Table 15

[0152] Example 6: Lipophilic delivery portion, modified nucleotide containing the same, and nucleic acid containing the modified nucleotide Certain abbreviations are defined as follows: "ACN" refers to acetonitrile, "AEX" refers to anion exchange, "C / D" refers to cleavage and deprotection, "CPG" refers to controlled pore glass, "aCSF" refers to artificial cerebrospinal fluid, "DCM" refers to dichloromethane, "DEA" refers to diethylamine, "DIPEA" refers to N,N-diisopropylethylamine, "DMA" refers to dimethylacetamide, "DMAP" refers to 4-dimethylaminopyridine, "DMF" refers to dimethylformamide, "DMSO" refers to dimethyl sulfoxide, and "DMT" refers to 4,4'-dimethoxytrityl(4,"4'-dimethoxytrityl" refers to 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, "ES / MS" refers to electrospray mass spectrometry, "siRNA" refers to ethyl acetate, "EtOH" refers to ethanol and ethyl alcohol, "IP-RP" refers to ion-pair reverse phase, "LC / MS" refers to liquid chromatography-mass spectrometry, "MeOH" refers to methanol and methyl alcohol, "MPA" refers to mobile phase A, "MPB" refers to mobile phase B, "MWCO (molecular weight cut-off)" refers to molecular weight cut-off, and "NaOAc" refers to sodium acetate. "N-hydroxysuccinimide" refers to trifluoroacetic acid, "NMR" refers to nuclear magnetic resonance, "PBS" refers to phosphate-buffered saline, "PVDF" refers to polyvinylidene fluoride, "RP" refers to reverse phase, "siRNA (small interfering ribonucleic acid)" refers to small interfering ribonucleic acid, "TCEP" refers to tris(2-carboxyethyl)phosphine, "TEA" refers to triethylamine, "TFA" refers to trifluoroacetic acid, "THF" refers to tetrahydrofuran, "UPLC" refers to ultra-performance liquid chromatography, and "UV" refers to ultraviolet light.

[0153] Scheme 1

[0154] [ka]

[0155] Scheme 1, step A shows the reaction of compound (1) with 2,2'-dipyridyl disulfide in a solvent system such as MeOH and THF to obtain compound (2). Step B shows the reaction of compound (2) with 3-sulfanylpropionic acid in a solvent such as MeOH to obtain compound (3). Step C shows the addition of NHS to compound (3) using a coupling reagent such as EDCI and a catalyst such as DMAP in a solvent such as DCM to obtain compound (4). Step D shows the addition of compound (4) to a properly modified sense chain in the presence of borate buffer to obtain compound (5).

[0156] Preparation 1 2-Dodecyldisulfaneyl pyridine

[0157] [ka] 1-Dodecanethiol (12.7 g, 61.4 mmol) was added to a solution of 2,2'-dipyridyl disulfide (20.5 g, 92.1 mmol) in MeOH (90 mL) and THF (5 mL). The mixture was stirred at ambient temperature for 16 hours and then concentrated under vacuum. The resulting residue was purified by silica gel flash chromatography eluted with 0-15% siRNA in hexane to obtain the title compound as a colorless oil (14.35 g, 75%). ES / MS (m / z): 312 (M+H).

[0158] Preparation 2 3-(dodecyldisulfanyl)propanoic acid

[0159] [ka] 3-Sulfanylpropionic acid (7.58 g, 71.44 mmol) was added to a solution of 2-(dodecyldisulfanyl)pyridine (18.55 g, 59.5 mmol) in MeOH (60 mL). The reaction mixture was stirred at ambient temperature for 1 hour and then concentrated under vacuum. The resulting residue was purified by silica gel flash chromatography eluted with 5-30% toluene in hexane to obtain the title compound as a colorless oil (14 g, 76%). 1 H NMR (DMSO-d6) δ2.86(t,2H,J=7.0Hz),2.71(t,2H,J=7.0Hz),2.62(t,2H,J=7.0Hz),1.61(quint,2H),1.33(q,2H),1.28(s,16H),0.90(t,3H,J=6.8Hz).

[0160] Preparation 3 2,5-Dioxopyrrolidine-1-yl-3-(dodecyldisulfanyl)propanoate

[0161] [ka] NHS (1.35 g, 11.7 mmol) was added to a solution of 3-(dodecyldisulfanyl)propanoic acid (3.0 g, 9.8 mmol), EDCI (2.25 g, 11.7 mmol), and DMAP (0.24 g, 2 mmol) in DCM (39 mL). The mixture was stirred at ambient temperature for 3 hours and then concentrated under vacuum. The resulting residue was purified by silica gel flash chromatography eluted with 0-40% phenylethylamine in hexane to obtain the title compound as a white solid (3.2 g, 81%). 1 H NMR(DMSO-d6)δ3.10(t,2H,J=6.3Hz),2.99(t,2H,J=6.3Hz),2.80(s,4H),2.75(t, 2H,J=7.0Hz),1.61(quint,2H),1.33(q,2H),1.28(s,16H),0.90(t,3H,J=6.8Hz).

[0162] C12 ADS-linked siRNA

[0163] [ka] Sense chains (3.1 g, 0.44 mmol) synthesized using the conditions found in the following protocol were treated with a solution of 2,5-dioxopyrrolidine-1-yl-3-(dodecyldisulfanyl)propanoate (5.3 g, 4.4 mmol) in ACN (113 mL). The solution was shaken at 30°C for 1.5 hours. The reaction was quenched by dilution with water and adjustment of pH to 7 with 1.2 M aqueous HCl. The solution was then concentrated via Genevac to remove the organic solvent and obtain crude oligonucleotides.

[0164] Crude oligonucleotides were purified using a reverse-phase AKTA® Pure purification system with a Source 15 RPC column (MPA: 50 mM NaOAc with 10% ACN and MPB: 80% ACN / water). In all cases, fractions with a mass purity of over 85% and free of impurities of over 5% were combined.

[0165] The purified oligonucleotide was desalted at 3500xg for approximately 30 minutes using a 15 mL 3K MWCO centrifuge spin tube. The oligonucleotide was rinsed with RNAse-free water until the conductivity of the eluate reached less than 100 usei / cm. After desalting was complete, 2–3 mL of RNAse-free water was added, followed by 10 aspirates, and the retained material was transferred to a 50 mL Falcon tube. This was repeated until the oligonucleotide was completely transferred by measuring the concentration of the compound on the filter via nanodrops. The final oligonucleotide was then nanofiltered twice at 3500xg for 2 minutes through a 15 mL 100K MWCO centrifuge spin tube. The final desalted oligonucleotide was analyzed for concentration (nanodrops at A260), characterized by mass purity by IP-RP LCMS, and UV purity by ULC. ES / MS (m / z): 7324.6 (M+H).

[0166] [Table 16-1]

[0167] [Table 16-2]

[0168] The delivery portions in Table 10 are synthesized and conjugated using standard methods. See, for example, International Publication No. 2019 / 217459.

[0169] Example 7: Tissue distribution and microgliosis analysis The selected SCN10A RNAi agents will be evaluated in rats for their tissue distribution and microgliosis. Rats will be intrathecally injected with a double-stranded RNAi agent (168 or 169) or PBS (phosphate-buffered saline). The animals will be sacrificed two months after injection.

[0170] RNAi agent tissue distribution and microgliosis analysis Selected tissues from the brain or spinal cord of treated rats are prepared for imaging. The rat tissue images are analyzed for inflammation based on the criteria in Table 11.

[0171] [Table 17]

Claims

1. An RNA interference (RNAi) agent comprising a sense strand and an antisense strand, wherein the antisense strand comprises a sequence complementary to at least 15 nucleotides of a sequence selected from any one of SEQ ID NOs: 2 to 32, the sense strand and the antisense strand form a double helix, optionally each independently comprises one or more modified nucleotides, and optionally one or more internucleotide bonds of the sense strand or the antisense strand are modified nucleotide bonds.

2. The RNAi agent according to claim 1, wherein the antisense strand comprises a sequence complementary to 18 consecutive nucleotides from any one of SEQ ID NOs: 2 to 32.

3. The RNAi agent according to claim 1 or 2, wherein the antisense strand comprises 3' overhangs of two nucleotides.

4. The RNAi agent according to claim 1, 2, or 3, wherein the antisense strand is 23 nucleotides long.

5. The RNAi agent according to any one of claims 1 to 4, wherein the sense strand is 21 nucleotides long.

6. The RNAi agent according to any one of claims 1 to 5, wherein the antisense strand comprises one sequence from sequence numbers 64 to 93.

7. The RNAi agent according to any one of claims 1 to 6, wherein the sense strand comprises one sequence from 33 to 63.

8. The RNAi agent according to claim 1, wherein the sense strand and the antisense strand form a double helix containing pairs of nucleic acid sequences as shown in Table 2.

9. The RNAi agent according to any one of claims 1 to 8, wherein the sense strand includes SEQ ID NO: 35 and the antisense strand includes SEQ ID NO: 66, or the sense strand includes SEQ ID NO: 39 and the antisense strand includes SEQ ID NO:

70.

10. The RNAi agent according to any one of claims 1 to 9, wherein the RNAi agent further comprises a delivery portion.

11. The RNAi agent according to claim 10, wherein the delivery portion is a lipophilic delivery portion.

12. The RNAi agent according to claim 11, wherein the lipophilic delivery portion is optionally of formula I or 2'-O-hexadecyl.

13. The RNAi agent according to any one of claims 1 to 12, wherein one or more of the modified nucleotides are independently 2'-fluoromodified nucleotides or 2'-O-methyl modified nucleotides.

14. The RNAi agent according to any one of claims 1 to 13, wherein each nucleotide of the sense strand and each nucleotide of the antisense strand are modified.

15. The antisense chain is 23 nucleotides long, each nucleotide in the antisense chain is a modified nucleotide, and one or more of the modified nucleotides are a. From the 5' end of the antisense chain, at positions 2, 3, 7, 14, and 16, b. From the 5' end of the antisense chain, at positions 2, 5, 7, 14, and 16, c. From the 5' end of the antisense chain, at positions 2, 3, 8, 14, and 16, d. From the 5' end of the antisense chain, at positions 2, 5, 8, 14, and 16, e. The 2nd, 14th, and 16th positions from the 5' end of the antisense chain, or f. The RNAi agent according to any one of claims 1 to 14, comprising a 2' fluoromodification located at a group of positions such as the 2, 6, 14, and 16 positions from the 5' end of the antisense strand.

16. The sense strand is 21 nucleotides long, each nucleotide in the sense strand is a modified nucleotide, and one or more of the modified nucleotides are a. From the 5' end of the sense strand, positions 9, 10, and 11, b. From the 5' end of the sense strand, positions 7, 9, and 11, c. From the 5' end of the sense strand, positions 7, 9, and 10, d. Positions 7, 10, and 11 from the 5' end of the sense strand, or e. The RNAi agent according to any one of claims 1 to 14, comprising a 2' fluoromodification located at a group of positions such as the 7th, 9th, 10th, and 11th positions from the 5' end of the sense strand.

17. The RNAi agent according to any one of claims 1 to 16, wherein the RNAi agent comprises one or more modified nucleotide links that are phosphorothioate chains.

18. The RNAi agent according to claim 17, wherein the sense strand has four or five phosphorothioate chains.

19. The RNAi agent according to claim 18, wherein the antisense strand has four or five phosphorothioate chains.

20. The RNAi agent according to any one of claims 1 to 19, wherein the nucleotide at the 5' end of the antisense strand has a phosphate group or a phosphate analogue.

21. The RNAi agent according to any one of claims 1 to 20, wherein the nucleotide at the 5' end of the antisense strand has a phosphate analog which is 5'-(E)-vinylphosphonate.

22. The RNAi agent according to any one of claims 1 to 21, wherein the sense strand has a modification that is a debase portion or further modification.

23. The RNAi agent according to any one of claims 1 to 22, wherein the nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand, and the nucleotides at positions 7, 9, 10, and 11 from the 5' end of the sense strand, each contain a 2' fluoromodification.

24. The RNAi agent according to any one of claims 1 to 22, wherein the nucleotides at positions 2, 5, 7, 14, and 16 from the 5' end of the antisense strand, and the nucleotides at positions 9, 10, and 11 from the 5' end of the sense strand, each contain a 2' fluoromodification.

25. The RNAi agent according to any one of claims 1 to 22, wherein the nucleotides at positions 2, 3, 7, 14, and 16 from the 5' end of the antisense strand, and the nucleotides at positions 9, 10, and 11 from the 5' end of the sense strand, each contain a 2' fluoromodification.

26. The RNAi agent according to any one of claims 1 to 22, wherein the nucleotides at positions 2, 5, 8, 14, and 16 from the 5' end of the antisense strand, and the nucleotides at positions 9, 10, and 11 from the 5' end of the sense strand, each contain a 2' fluoromodification.

27. The RNAi agent according to any one of claims 1 to 22, wherein the nucleotides at positions 2, 14, and 16 from the 5' end of the antisense strand, and the nucleotides at positions 9, 10, and 11 from the 5' end of the sense strand, each contain a 2' fluoromodification.

28. The RNAi agent according to any one of claims 1 to 22, wherein the nucleotides at positions 2, 5, 7, 14, and 16 from the 5' end of the antisense strand, and the nucleotides at positions 7, 9, 10, and 11 from the 5' end of the sense strand, each contain a 2' fluoromodification.

29. The RNAi agent according to any one of claims 10 to 28, wherein the delivery portion is a lipophilic delivery portion conjugated to a nucleotide at position 12 or 13 from the 5' end of the sense strand.

30. The RNAi agent according to any one of claims 10 to 29, wherein the delivery portion is a lipophilic delivery portion conjugated to a nucleotide at the 12th or 13th position from the 5' end of the sense strand, and the lipophilic delivery portion is 2'-O-hexadecyl.

31. The RNAi agent according to any one of claims 10 to 29, wherein the delivery portion is a lipophilic delivery portion conjugated to a nucleotide at the 12th or 13th position from the 5' end of the sense strand, and the lipophilic delivery portion has formula I.

32. The sense chain and the antisense chain form a double chain. a. The sense strand has at least 90% sequence identity with sequence number 33, and the antisense strand has at least 90% sequence identity with sequence number 64, b. The sense strand has at least 90% sequence identity with sequence number 34, and the antisense strand has at least 90% sequence identity with sequence number 65, c. The sense strand has at least 90% sequence identity with sequence number 35, and the antisense strand has at least 90% sequence identity with sequence number 66, d. The sense strand has at least 90% sequence identity with sequence number 36, and the antisense strand has at least 90% sequence identity with sequence number 67, e. The sense strand has at least 90% sequence identity with sequence number 37, and the antisense strand has at least 90% sequence identity with sequence number 68, f. The sense strand has at least 90% sequence identity with sequence number 38, and the antisense strand has at least 90% sequence identity with sequence number 69, g. The sense strand has at least 90% sequence identity with sequence number 39, and the antisense strand has at least 90% sequence identity with sequence number 70, h. The sense strand has at least 90% sequence identity with sequence number 40, and the antisense strand has at least 90% sequence identity with sequence number 71, i. The sense strand has at least 90% sequence identity with sequence number 41, and the antisense strand has at least 90% sequence identity with sequence number 72, j. The sense strand has at least 90% sequence identity with sequence number 42, and the antisense strand has at least 90% sequence identity with sequence number 73, k. The sense strand has at least 90% sequence identity with sequence number 43, and the antisense strand has at least 90% sequence identity with sequence number 74, l. The sense strand has at least 90% sequence identity with sequence number 44, and the antisense strand has at least 90% sequence identity with sequence number 75, m. The sense strand has at least 90% sequence identity with sequence number 45, and the antisense strand has at least 90% sequence identity with sequence number 76, n. The sense strand has at least 90% sequence identity with sequence number 46, and the antisense strand has at least 90% sequence identity with sequence number 77, o. The sense strand has at least 90% sequence identity with sequence number 47, and the antisense strand has at least 90% sequence identity with sequence number 78, p. The sense strand has at least 90% sequence identity with sequence number 48, and the antisense strand has at least 90% sequence identity with sequence number 79, q. The sense strand has at least 90% sequence identity with sequence number 49, and the antisense strand has at least 90% sequence identity with sequence number 80, r. The sense strand has at least 90% sequence identity with sequence number 50, and the antisense strand has at least 90% sequence identity with sequence number 81, s. The sense strand has at least 90% sequence identity with sequence number 51, and the antisense strand has at least 90% sequence identity with sequence number 82, t. The sense strand has at least 90% sequence identity with sequence number 52, and the antisense strand has at least 90% sequence identity with sequence number 83, u. The sense strand has at least 90% sequence identity with sequence number 53, and the antisense strand has at least 90% sequence identity with sequence number 84, v. The sense strand has at least 90% sequence identity with sequence number 54, and the antisense strand has at least 90% sequence identity with sequence number 85, w. The sense strand has at least 90% sequence identity with sequence number 55, and the antisense strand has at least 90% sequence identity with sequence number 86, x. The sense strand has at least 90% sequence identity with sequence number 56, and the antisense strand has at least 90% sequence identity with sequence number 87, y. The sense strand has at least 90% sequence identity with sequence number 57, and the antisense strand has at least 90% sequence identity with sequence number 88, z. The sense strand has at least 90% sequence identity with sequence number 58, and the antisense strand has at least 90% sequence identity with sequence number 89, aa. The sense strand has at least 90% sequence identity with sequence number 59, and the antisense strand has at least 90% sequence identity with sequence number 90, bb. The sense strand has at least 90% sequence identity with sequence number 60, and the antisense strand has at least 90% sequence identity with sequence number 91, cc. The sense strand has at least 90% sequence identity with sequence number 61, and the antisense strand has at least 90% sequence identity with sequence number 92, dd. The sense strand has at least 90% sequence identity with SEQ ID NO: 62, and the antisense strand has at least 90% sequence identity with SEQ ID NO: 93, or ee. The RNAi agent according to any one of claims 1 to 7 and 10 to 31, wherein the sense strand has at least 90% sequence identity with SEQ ID NO: 63, and the antisense strand has at least 90% sequence identity with SEQ ID NO:

66.

33. a. The sense chain includes sequence number 94, and the antisense chain includes sequence number 132. b. The sense chain includes sequence number 95, and the antisense chain includes sequence number 133. c. The sense chain includes sequence number 96, and the antisense chain includes sequence number 134. d. The sense chain includes sequence number 97, and the antisense chain includes sequence number 135. e. The sense chain includes sequence number 98, and the antisense chain includes sequence number 136. f. The sense chain includes sequence number 99, and the antisense chain includes sequence number 137, g. The sense chain includes sequence number 100, and the antisense chain includes sequence number 138. h. The sense chain includes sequence number 101, and the antisense chain includes sequence number 139, i. The sense chain includes sequence number 102, and the antisense chain includes sequence number 140. j. The sense chain includes sequence number 103, and the antisense chain includes sequence number 141. k. The sense chain includes sequence number 104, and the antisense chain includes sequence number 142. l. The sense chain includes sequence number 105, and the antisense chain includes sequence number 143. m. The sense chain includes sequence number 106, and the antisense chain includes sequence number 144. n. The sense chain includes sequence number 107, and the antisense chain includes sequence number 145. o. The sense chain includes sequence number 108, and the antisense chain includes sequence number 146. p. The sense chain includes sequence number 109, and the antisense chain includes sequence number 147. q. The sense chain includes sequence number 110, and the antisense chain includes sequence number 148. r. The sense chain includes sequence number 111, and the antisense chain includes sequence number 149. s. The sense chain includes sequence number 112, and the antisense chain includes sequence number 150. t. The sense chain includes sequence number 113, and the antisense chain includes sequence number 151. u. The sense chain includes sequence number 114, and the antisense chain includes sequence number 152. v. The sense chain includes sequence number 115, and the antisense chain includes sequence number 153, w. The sense chain includes sequence number 116, and the antisense chain includes sequence number 154. x. The sense chain includes sequence number 117, and the antisense chain includes sequence number 155. y. The sense chain includes sequence number 118, and the antisense chain includes sequence number 156. z. The sense chain includes sequence number 119, and the antisense chain includes sequence number 157, aa. The sense chain includes sequence number 120, and the antisense chain includes sequence number 158. bb. The sense chain includes sequence number 121, and the antisense chain includes sequence number 159. cc. The sense chain includes sequence number 122, and the antisense chain includes sequence number 160. dd. The sense chain includes sequence number 123, and the antisense chain includes sequence number 161. ee. The sense chain includes sequence number 96, and the antisense chain includes sequence number 162. ff. The sense chain includes sequence number 96, and the antisense chain includes sequence number 163. gg. The sense chain includes sequence number 96, and the antisense chain includes sequence number 164. hh. The sense chain includes sequence number 96, and the antisense chain includes sequence number 165. ii. The sense chain includes sequence number 96, and the antisense chain includes sequence number 166. jj. The sense chain includes sequence number 124, and the antisense chain includes sequence number 162, kk. The sense chain includes sequence number 124, and the antisense chain includes sequence number 163. ll. The sense chain includes sequence number 124, and the antisense chain includes sequence number 164. mm. The sense chain includes sequence number 124, and the antisense chain includes sequence number 165. nn. The sense chain includes sequence number 124, and the antisense chain includes sequence number 166. oo. The sense chain includes sequence number 124, and the antisense chain includes sequence number 134. pp. The sense chain includes sequence number 125, and the antisense chain includes sequence number 162. qq. The sense chain includes sequence number 125, and the antisense chain includes sequence number 163. rr. The sense chain includes sequence number 125, and the antisense chain includes sequence number 164. ss. The sense chain includes sequence number 125, and the antisense chain includes sequence number 165. tt. The sense chain includes sequence number 125, and the antisense chain includes sequence number 166. uu. The sense chain includes sequence number 125, and the antisense chain includes sequence number 134. vv. The sense chain includes sequence number 126, and the antisense chain includes sequence number 162, LOL. The sense chain includes sequence number 126, and the antisense chain includes sequence number 163. xx. The sense chain includes sequence number 126, and the antisense chain includes sequence number 164. yy. The sense chain includes sequence number 126, and the antisense chain includes sequence number 165. zz. The sense chain includes sequence number 126, and the antisense chain includes sequence number 166. aaa. The sense chain includes sequence number 126, and the antisense chain includes sequence number 134. bbb. The sense chain includes sequence number 127, and the antisense chain includes sequence number 167. ccc. The sense chain includes sequence number 127, and the antisense chain includes sequence number 168, ddd. The sense chain includes sequence number 127, and the antisense chain includes sequence number 169. eee. The sense chain includes sequence number 128, and the antisense chain includes sequence number 170. fff. The sense chain includes sequence number 128, and the antisense chain includes sequence number 171, ggg. The sense chain includes sequence number 128, and the antisense chain includes sequence number 172, hhh. The sense chain includes sequence number 129, and the antisense chain includes sequence number 173. iii. The sense chain includes sequence number 129, and the antisense chain includes sequence number 174. jjj. The sense chain includes sequence number 129, and the antisense chain includes sequence number 175, kkk. The sense chain includes sequence number 130, and the antisense chain includes sequence number 176. lll. The sense chain includes sequence number 130, and the antisense chain includes sequence number 177, mmm. The sense chain includes sequence number 130, and the antisense chain includes sequence number 178. nnn. The sense chain includes sequence number 131, and the antisense chain includes sequence number 179, oo. The sense chain includes sequence number 131, and the antisense chain includes sequence number 180. ppp. The sense chain includes sequence number 131, and the antisense chain includes sequence number 181, qqq. The sense chain includes sequence number 232, and the antisense chain includes sequence number 233. rrr. The sense chain includes sequence number 234, and the antisense chain includes sequence number 235, sss. The sense chain includes sequence number 236, and the antisense chain includes sequence number 233. ttt. The sense chain includes sequence number 237, and the antisense chain includes sequence number 233. uuu. The sense chain includes sequence number 238, and the antisense chain includes sequence number 239. vvv. The sense chain includes sequence number 240, and the antisense chain includes sequence number 239, LOL. The sense chain includes sequence number 238, and the antisense chain includes sequence number 235. xxx. The sense chain includes sequence number 240, and the antisense chain includes sequence number 235. yyy. The sense chain includes sequence number 241, and the antisense chain includes sequence number 235. zzz. The sense chain includes sequence number 238, and the antisense chain includes sequence number 242. aaaa. The sense chain includes sequence number 232, and the antisense chain includes sequence number 239, bbbb. The sense chain includes sequence number 240, and the antisense chain includes sequence number 242. cccc. The sense chain includes sequence number 94, and the antisense chain includes sequence number 182. dddd. The sense chain includes sequence number 95, and the antisense chain includes sequence number 183. eeee. The sense chain includes sequence number 96, and the antisense chain includes sequence number 184. ffff. The sense chain includes sequence number 97, and the antisense chain includes sequence number 185. gggg. The sense chain includes sequence number 98, and the antisense chain includes sequence number 186. hhhh. The sense chain includes sequence number 99, and the antisense chain includes sequence number 187, iii. The sense chain includes sequence number 100, and the antisense chain includes sequence number 188. jjjj. The sense chain includes sequence number 101, and the antisense chain includes sequence number 189, kkkk. The sense chain includes sequence number 102, and the antisense chain includes sequence number 190. llll. The sense chain includes sequence number 103, and the antisense chain includes sequence number 191. mmmm. The sense chain includes sequence number 104, and the antisense chain includes sequence number 192. nnnn. The sense chain includes sequence number 105, and the antisense chain includes sequence number 193. oooo. The sense chain includes sequence number 106, and the antisense chain includes sequence number 194. pppp. The sense chain includes sequence number 107, and the antisense chain includes sequence number 195, qqqq. The sense chain includes sequence number 108, and the antisense chain includes sequence number 196. rrrr. The sense chain includes sequence number 109, and the antisense chain includes sequence number 197, sss. The sense chain includes sequence number 110, and the antisense chain includes sequence number 198. tttt. The sense chain includes sequence number 111, and the antisense chain includes sequence number 199, uu uu. The sense chain includes sequence number 112, and the antisense chain includes sequence number 200. vvvv. The sense chain includes sequence number 113, and the antisense chain includes sequence number 201, LOL. The sense chain includes sequence number 114, and the antisense chain includes sequence number 202. xxxx. The sense chain includes sequence number 115, and the antisense chain includes sequence number 203. yyyy. The sense chain includes sequence number 116, and the antisense chain includes sequence number 204. zzz. The sense chain includes sequence number 117, and the antisense chain includes sequence number 205. aaaaa. The sense chain includes sequence number 118, and the antisense chain includes sequence number 206, bbbbb. The sense chain includes sequence number 119, and the antisense chain includes sequence number 207, cccccc. The sense chain includes sequence number 120, and the antisense chain includes sequence number 208. dddddd. The sense chain includes sequence number 121, and the antisense chain includes sequence number 209, eeee. The sense chain includes sequence number 122, and the antisense chain includes sequence number 210. fffff. The sense chain includes sequence number 123, and the antisense chain includes sequence number 211. ggggg. The sense chain includes sequence number 96, and the antisense chain includes sequence number 212. hhhhh. The sense chain includes sequence number 96, and the antisense chain includes sequence number 213. iiiiii. The sense chain includes sequence number 96, and the antisense chain includes sequence number 214. jjjjj. The sense chain includes sequence number 96, and the antisense chain includes sequence number 215. kkkkk. The sense chain includes sequence number 96, and the antisense chain includes sequence number 216. lllll. The sense chain includes sequence number 124, and the antisense chain includes sequence number 212, mmmmm. The sense chain includes sequence number 124, and the antisense chain includes sequence number 213. nnnnn. The sense chain includes sequence number 124, and the antisense chain includes sequence number 214. oooooo. The sense chain includes sequence number 124, and the antisense chain includes sequence number 215. ppppp. The sense chain includes sequence number 124, and the antisense chain includes sequence number 216, qqqqq. The sense chain includes sequence number 124, and the antisense chain includes sequence number 184. rrrrr. The sense chain includes sequence number 125, and the antisense chain includes sequence number 212, ssss. The sense chain includes sequence number 125, and the antisense chain includes sequence number 213. ttttt. The sense chain includes sequence number 125, and the antisense chain includes sequence number 214. uuuuu. The sense chain includes sequence number 125, and the antisense chain includes sequence number 215. vvvvv. The sense chain includes sequence number 125, and the antisense chain includes sequence number 216, LOL. The sense chain includes sequence number 125, and the antisense chain includes sequence number 184. xxxxx. The sense chain includes sequence number 126, and the antisense chain includes sequence number 212. yyyyy. The sense chain includes sequence number 126, and the antisense chain includes sequence number 213. zzzzz. The sense chain includes sequence number 126, and the antisense chain includes sequence number 214. aaaaaa. The sense chain includes sequence number 126, and the antisense chain includes sequence number 215. bbbbbb. The sense chain includes sequence number 126, and the antisense chain includes sequence number 216. cccccc. The sense chain includes sequence number 126, and the antisense chain includes sequence number 184. dddddd. The sense chain includes sequence number 127, and the antisense chain includes sequence number 217, eeeeee. The sense chain includes sequence number 127, and the antisense chain includes sequence number 218. fffff. The sense chain includes sequence number 127, and the antisense chain includes sequence number 219, gggggg. The sense chain includes sequence number 128, and the antisense chain includes sequence number 220, hhhhhh. The sense chain includes sequence number 128, and the antisense chain includes sequence number 221. iiiiii. The sense chain includes sequence number 128, and the antisense chain includes sequence number 222. jjjjjj. The sense chain includes sequence number 129, and the antisense chain includes sequence number 223. kkkkkk. The sense chain includes sequence number 129, and the antisense chain includes sequence number 224. lllllll. The sense chain includes sequence number 129, and the antisense chain includes sequence number 225, mmmmmm. The sense chain includes sequence number 130, and the antisense chain includes sequence number 226. nnnnnnn. The sense chain includes sequence number 130, and the antisense chain includes sequence number 227, oooooo. The sense chain includes sequence number 130, and the antisense chain includes sequence number 228. pppppp. The sense chain includes sequence number 131, and the antisense chain includes sequence number 229, qqqqqq. The sense chain includes sequence number 131, and the antisense chain includes sequence number 230. rrrrrr. The sense chain includes sequence number 131, and the antisense chain includes sequence number 231, sssss. The sense chain includes sequence number 232, and the antisense chain includes sequence number 184. ttttttt. The sense chain includes sequence number 234, and the antisense chain includes sequence number 215. uuuuuu. The sense chain includes sequence number 236, and the antisense chain includes sequence number 184. vvvvvv. The sense chain includes sequence number 237, and the antisense chain includes sequence number 184, LOL. The sense chain includes sequence number 238, and the antisense chain includes sequence number 213. xxxxxx. The sense chain includes sequence number 240, and the antisense chain includes sequence number 213. yyyyyy. The sense chain includes sequence number 238, and the antisense chain includes sequence number 215. zzzzzzz. The sense chain includes sequence number 240, and the antisense chain includes sequence number 215. aaaaaaa. The sense chain includes sequence number 241, and the antisense chain includes sequence number 215, bbbbbb. The sense chain includes sequence number 238, and the antisense chain includes sequence number 212. cccccccc. The sense chain includes sequence number 232, and the antisense chain includes sequence number 213. dddddddd. The RNAi agent according to any one of claims 1 to 7 and 9 to 31, wherein the sense strand comprises SEQ ID NO: 240 and the antisense strand comprises SEQ ID NO:

214.

34. a. The sense chain consists of sequence number 94, and the antisense chain consists of sequence number 132. b. The sense chain consists of sequence number 95, and the antisense chain consists of sequence number 133. c. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 134. d. The sense chain consists of sequence number 97, and the antisense chain consists of sequence number 135. e. The sense chain consists of sequence number 98, and the antisense chain consists of sequence number 136. f. The sense chain consists of sequence number 99, and the antisense chain consists of sequence number 137. g. The sense chain consists of sequence number 100, and the antisense chain consists of sequence number 138. h. The sense chain consists of sequence number 101, and the antisense chain consists of sequence number 139. i. The sense chain consists of sequence number 102, and the antisense chain consists of sequence number 140, j. The sense chain consists of sequence number 103, and the antisense chain consists of sequence number 141. k. The sense chain consists of sequence number 104, and the antisense chain consists of sequence number 142. l. The sense chain consists of sequence number 105, and the antisense chain consists of sequence number 143. m. The sense chain consists of sequence number 106, and the antisense chain consists of sequence number 144. n. The sense chain consists of sequence number 107, and the antisense chain consists of sequence number 145. o. The sense chain consists of sequence number 108, and the antisense chain consists of sequence number 146. p. The sense chain consists of sequence number 109, and the antisense chain consists of sequence number 147. q. The sense chain consists of sequence number 110, and the antisense chain consists of sequence number 148, r. The sense chain consists of sequence number 111, and the antisense chain consists of sequence number 149, s. The sense chain consists of sequence number 112, and the antisense chain consists of sequence number 150. t. The sense chain consists of sequence number 113, and the antisense chain consists of sequence number 151. u. The sense chain consists of sequence number 114, and the antisense chain consists of sequence number 152. v. The sense chain consists of sequence number 115, and the antisense chain consists of sequence number 153, w. The sense chain consists of sequence number 116, and the antisense chain consists of sequence number 154. x. The sense chain consists of sequence number 117, and the antisense chain consists of sequence number 155. y. The sense chain consists of sequence number 118, and the antisense chain consists of sequence number 156. z. The sense chain consists of sequence number 119, and the antisense chain consists of sequence number 157. aa. The sense chain consists of sequence number 120, and the antisense chain consists of sequence number 158. bb. The sense chain consists of sequence number 121, and the antisense chain consists of sequence number 159. cc. The sense chain consists of sequence number 122, and the antisense chain consists of sequence number 160. dd. The sense chain consists of sequence number 123, and the antisense chain consists of sequence number 161. ee. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 162. ff. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 163. gg. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 164. hh. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 165. ii. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 166. jj. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 162, kk. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 163. ll. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 164. mm. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 165. nn. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 166. oo. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 134. pp. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 162. qq. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 163. rr. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 164, ss. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 165. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 166. uu. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 134. vv. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 162, LOL. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 163. xx. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 164. yy. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 165. zz. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 166. aaa. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 134. bbb. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 167. ccc. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 168. ddd. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 169. eee. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 170. fff. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 171. ggg. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 172. hhh. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 173. iii. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 174. jjj. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 175, kkk. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 176. lll. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 177, mmm. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 178. nnn. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 179, oo. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 180. ppp. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 181, qqq. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 233. rrr. The sense chain consists of sequence number 234, and the antisense chain consists of sequence number 235, sss. The sense chain consists of sequence number 236, and the antisense chain consists of sequence number 233. The sense chain consists of sequence number 237, and the antisense chain consists of sequence number 233. uuu. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 239. vvv. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 239, LOL. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 235. xxx. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 235. yyy. The sense chain consists of sequence number 241, and the antisense chain consists of sequence number 235. zzz. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 242. aaaa. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 239, bbbb. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 242. cccc. The sense chain consists of sequence number 94, and the antisense chain consists of sequence number 182. dddd. The sense chain consists of sequence number 95, and the antisense chain consists of sequence number 183. eeee. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 184. fff. The sense chain consists of sequence number 97, and the antisense chain consists of sequence number 185. gggg. The sense chain consists of sequence number 98, and the antisense chain consists of sequence number 186. hhhh. The sense chain consists of sequence number 99, and the antisense chain consists of sequence number 187. iii. The sense chain consists of sequence number 100, and the antisense chain consists of sequence number 188. jjjj. The sense chain consists of sequence number 101, and the antisense chain consists of sequence number 189, kkkk. The sense chain consists of sequence number 102, and the antisense chain consists of sequence number 190. llll. The sense chain consists of sequence number 103, and the antisense chain consists of sequence number 191. mmmm. The sense chain consists of sequence number 104, and the antisense chain consists of sequence number 192. nnnn. The sense chain consists of sequence number 105, and the antisense chain consists of sequence number 193, oooo. The sense chain consists of sequence number 106, and the antisense chain consists of sequence number 194. pppp. The sense chain consists of sequence number 107, and the antisense chain consists of sequence number 195, qqqq. The sense chain consists of sequence number 108, and the antisense chain consists of sequence number 196. rrrr. The sense chain consists of sequence number 109, and the antisense chain consists of sequence number 197, sss. The sense chain consists of sequence number 110, and the antisense chain consists of sequence number 198. tttt. The sense chain consists of sequence number 111, and the antisense chain consists of sequence number 199, uuuu. The sense chain consists of sequence number 112, and the antisense chain consists of sequence number 200. vvvv. The sense chain consists of sequence number 113, and the antisense chain consists of sequence number 201, LOL. The sense chain consists of sequence number 114, and the antisense chain consists of sequence number 202. xxxx. The sense chain consists of sequence number 115, and the antisense chain consists of sequence number 203. yyyy. The sense chain consists of sequence number 116, and the antisense chain consists of sequence number 204. zzz. The sense chain consists of sequence number 117, and the antisense chain consists of sequence number 205. aaaaa. The sense chain consists of sequence number 118, and the antisense chain consists of sequence number 206, bbbbb. The sense chain consists of sequence number 119, and the antisense chain consists of sequence number 207. cccccc. The sense chain consists of sequence number 120, and the antisense chain consists of sequence number 208. dddddd. The sense chain consists of sequence number 121, and the antisense chain consists of sequence number 209, eeee. The sense chain consists of sequence number 122, and the antisense chain consists of sequence number 210. fffff. The sense chain consists of sequence number 123, and the antisense chain consists of sequence number 211. ggggg. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 212. hhhhh. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 213. iiiiii. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 214, jjjjj. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 215. kkkkk. The sense chain consists of sequence number 96, and the antisense chain consists of sequence number 216. lllll. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 212, mmmmm. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 213. nnnnn. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 214, oooooo. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 215. ppppp. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 216, qqqqq. The sense chain consists of sequence number 124, and the antisense chain consists of sequence number 184. rrrrr. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 212, ssss. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 213. ttttt. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 214. uuuuu. The sense chain includes sequence number 125, and the antisense chain includes sequence number 215. vvvvv. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 216, LOL. The sense chain consists of sequence number 125, and the antisense chain consists of sequence number 184. xxxxx. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 212. yyyyy. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 213. zzzzz. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 214. aaaaaa. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 215, bbbbbb. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 216. cccccc. The sense chain consists of sequence number 126, and the antisense chain consists of sequence number 184. dddddd. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 217, eeeeee. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 218. fffff. The sense chain consists of sequence number 127, and the antisense chain consists of sequence number 219, gggggg. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 220. hhhhhh. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 221. iiiiiii. The sense chain consists of sequence number 128, and the antisense chain consists of sequence number 222, jjjjjj. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 223, kkkkkk. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 224. lllllll. The sense chain consists of sequence number 129, and the antisense chain consists of sequence number 225, mmmmmm. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 226. nnnnnn. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 227, oooooo. The sense chain consists of sequence number 130, and the antisense chain consists of sequence number 228. ppppp. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 229, qqqqqq. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 230. rrrrrr. The sense chain consists of sequence number 131, and the antisense chain consists of sequence number 231, sssss. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 184. ttttttt. The sense chain consists of sequence number 234, and the antisense chain consists of sequence number 215. uuuuuu. The sense chain consists of sequence number 236, and the antisense chain consists of sequence number 184. vvvvvv. The sense chain consists of sequence number 237, and the antisense chain consists of sequence number 184, LOL. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 213. xxxxxx. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 213. yyyyy. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 215. zzzzzzz. The sense chain consists of sequence number 240, and the antisense chain consists of sequence number 215. aaaaaaa. The sense chain consists of sequence number 241, and the antisense chain consists of sequence number 215, bbbbbbbb. The sense chain consists of sequence number 238, and the antisense chain consists of sequence number 212. cccccccc. The sense chain consists of sequence number 232, and the antisense chain consists of sequence number 213. dddddddd. The RNAi agent according to claim 33, wherein the sense strand consists of Sequence ID No. 240 and the antisense strand consists of Sequence ID No.

214.

35. The RNAi agent according to claim 33, comprising a delivery portion, wherein the delivery portion is a lipophilic delivery portion conjugated to a nucleotide at position 12 or 13 from the 5' end of the sense strand.

36. The RNAi agent according to claim 35, wherein the delivery portion is a lipophilic delivery portion conjugated to a nucleotide at the 12th or 13th position from the 5' end of the sense strand, and the lipophilic delivery portion is 2'-O-hexadecyl.

37. The RNAi agent according to any one of claims 1 to 19, wherein the delivery portion is a lipophilic delivery portion conjugated to a nucleotide at the 12th or 13th position from the 5' end of the sense strand, and the lipophilic delivery portion has formula I.

38. a. The sense chain includes sequence number 35 and the antisense chain includes sequence number 66, or b. The sense chain includes sequence number 39, and the antisense chain includes sequence number 70. The RNAi agent according to claim 1, wherein the first nucleotide from the 5' end of the antisense strand is a modified nucleotide having a phosphate analog, optionally the phosphate analog being a 5'-vinylphosphonate, the sense strand and the antisense strand have one, two, three, or four modified nucleotide interlinks, optionally the modified nucleotide interlinks being phosphorothioate chains, each nucleotide of the sense strand and the antisense strand is a modified nucleotide, the modified nucleotide of the antisense strand is a 2'-fluoromodified nucleotide at positions 2, 6, 14, and 16 from the 5' end of the antisense strand, and a 2'-O-methyl modified nucleotide at the remaining positions, the modified nucleotide of the sense strand is a 2'-fluoromodified nucleotide at positions 7, 9, 10, and 11 from the 5' end of the sense strand, a 2'-O-C16 alkyl modified nucleotide at position 13 from the 5' end of the sense strand, and a 2'-O-methyl modified nucleotide at the remaining positions.

39. a. The sense chain includes sequence number 35 and the antisense chain includes sequence number 66, or b. The sense chain includes sequence number 39, and the antisense chain includes sequence number 70. The RNAi agent according to claim 1, wherein the first nucleotide from the 5' end of the antisense strand is a modified nucleotide having a phosphate analog, optionally the phosphate analog being a 5'-vinylphosphonate, the sense strand and the antisense strand have one, two, three, or four modified nucleotide interlinks, optionally the modified nucleotide interlinks being phosphorothioate chains, each nucleotide of the sense strand and the antisense strand is a modified nucleotide, the modified nucleotide of the antisense strand is a 2'-fluoromodified nucleotide at positions 2, 6, 14, and 16 from the 5' end of the antisense strand, and a 2'-O-methyl modified nucleotide at the remaining positions, the modified nucleotide of the sense strand is a 2'-fluoromodified nucleotide at positions 7, 9, 10, and 11 from the 5' end of the sense strand, a 2'-O-C16 alkyl modified nucleotide at position 12 from the 5' end of the sense strand, and a 2'-O-methyl modified nucleotide at the remaining positions.

40. a. The sense chain includes sequence number 35 and the antisense chain includes sequence number 66, or b. The sense chain includes sequence number 39, and the antisense chain includes sequence number 70. The RNAi agent according to claim 1, wherein the first nucleotide from the 5' end of the antisense strand is a modified nucleotide having a phosphate analog, optionally the phosphate analog being a 5'-vinylphosphonate, the sense strand and the antisense strand have one, two, three, or four modified nucleotide interlinks, optionally the modified nucleotide interlinks being phosphorothioate chains, each nucleotide of the sense strand and the antisense strand is a modified nucleotide, the modified nucleotide of the antisense strand is a 2'-fluoromodified nucleotide at positions 2, 5, 8, 14, and 16 from the 5' end of the antisense strand, and a 2'-O-methylmodified nucleotide at the remaining positions, the modified nucleotide of the sense strand is a 2'-fluoromodified nucleotide at positions 9, 10, and 11 from the 5' end of the sense strand, a nucleotide of formula Ib at position 12 from the 5' end of the sense strand, and a 2'-O-methylmodified nucleotide at the remaining positions.

41. The RNAi agent according to claim 1, wherein the antisense strand essentially consists of SEQ ID NO: 233 and the sense strand essentially consists of SEQ ID NO:

232.

42. The RNAi agent according to claim 1, wherein the antisense strand essentially consists of Sequence ID No. 235 and the sense strand essentially consists of Sequence ID No.

234.

43. The RNAi agent according to claim 1, wherein the antisense strand essentially consists of SEQ ID NO: 233 and the sense strand essentially consists of SEQ ID NO:

236.

44. An RNAi agent according to any one of claims 1 to 43, for use in therapy.

45. An RNAi agent according to any one of claims 1 to 43, for use in the treatment of pain, or optionally, chronic pain.

46. Use of an RNAi agent according to any one of claims 1 to 43 for the manufacture of a pharmaceutical product for the treatment of pain, or optionally for the treatment of chronic pain.

47. A pharmaceutical composition comprising an RNAi agent according to any one of claims 1 to 43 and one or more pharmaceutically acceptable excipients.

48. A method for treating pain in a patient who requires treatment of pain, comprising administering an RNAi agent or a pharmaceutical composition thereof according to any one of claims 1 to 43.

49. The method according to claim 48, wherein the RNAi agent or pharmaceutical composition thereof according to any one of claims 1 to 43 is administered intrathecally or epidurally.

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