Rnai agent for inhibiting MUC5ac gene expression and use thereof

Abstract

Provided are an RNAi agent for inhibiting MUC5AC gene expression and the use thereof. Specifically disclosed are a double-stranded RNA for inhibiting MUC5AC gene expression, a vector, cell, pharmaceutical composition and kit containing a nucleotide encoding the dsRNA, and a method for treating diseases or conditions mediated by or associated with MUC5AC expression in a subject by using the dsRNA, vector, cell, or pharmaceutical composition.

Description

An RNAi agent that inhibits MUC5AC gene expression and its application

[0001] This application claims priority to Chinese Patent Application No. 2024119428274, filed on December 26, 2024, and Chinese Patent Application No. 2025119129385, filed on December 17, 2025, the entire contents of each of which are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of RNA interference. Background Technology

[0003] Airway mucus, as a crucial component of the airway's first line of defense, plays a vital role in maintaining airway humidification and assisting in the function of airway epithelial cells. However, when the respiratory tract is stimulated by pathogenic microorganisms or physical and chemical factors, it triggers the recruitment of inflammatory cells and the release of inflammatory mediators in the lungs, activating multiple signaling pathways and transcription factors, leading to increased airway mucus secretion. Long-term excessive mucus secretion (also known as chronic airway hypersecretion) causes mucus to accumulate in the airway lumen, obstructing the airway, restricting airflow, and accelerating the decline in lung function. Simultaneously, the inflammatory response reduces ciliary clearance function, depletes alveolar surfactant, and alters the biophysical properties of mucus, leading to repeated airway infections, obstruction, and remodeling, creating a vicious cycle. Studies have found that chronic airway hypersecretion is an important pathophysiological and clinical manifestation of mucooblocclusion-related airway diseases such as bronchial asthma and bronchiectasis.

[0004] Mucins are important components of airway mucus. MUC5AC and MUC5B are the main gel-forming mucins in airway mucus. Under normal circumstances, MUC5B is dominant, with its content being about 10 times that of MUC5AC, and it plays a natural defense role against various inhaled foreign substances. However, under the stimulation of exogenous stimuli, pathogenic microorganisms, and cigarette smoke, the expression levels of both MUC5AC and MUC5B mucins increase significantly, especially MUC5AC, which shows an abnormally high increase. The high expression of MUC5AC is closely related to excessive airway mucus secretion.

[0005] Currently, the main medications used clinically to treat mucus obstructive airway diseases include bronchodilators, glucocorticoids, phosphodiesterase inhibitors, anticholinergics, and expectorants. Existing treatments primarily focus on symptom relief and cannot directly address the pathogenic mucin overexpression and excessive secretion. Furthermore, all of these medications have varying degrees of toxic side effects; for example, high-dose bronchodilators can cause cardiovascular-related toxicity, while long-term use of hormones can lead to immunosuppression and osteoporosis. In addition, existing treatments require frequent inhalation, oral administration, or injection, resulting in low patient compliance. Therefore, there is an urgent need to find next-generation treatments that inhibit airway mucus hypersecretion.

[0006] The emergence of RNA interference (RNAi) technology has offered new possibilities for treating such diseases. RNAi technology utilizes double-stranded RNA (dsRNA) to specifically degrade or inhibit the expression of homologous mRNA, thereby blocking the expression of target genes. Currently, RNAi drugs can be administered once every six months, significantly reducing the frequency of dosing and thus improving patient compliance. Therefore, developing a highly effective RNAi interference agent to silence MUC5AC, fundamentally blocking airway hypersecretion, will provide an effective means for long-term treatment of mucus obstructive airway diseases. RNA-based therapeutics are known to inhibit or reduce MUC5AC gene expression. International patent applications WO2022251394A1 and WO2010067882A1 describe the construction of double-stranded RNA (dsRNA) for inhibiting or reducing MUC5AC expression, and the use of the constructs to treat asthma and obstructive pulmonary diseases. Furthermore, international patent application WO2023105527A1 describes antisense oligonucleotides for inhibiting or reducing MUC5AC expression. Although some RNAi drugs targeting MUC5AC are under development, none are currently on the market.

[0007] Therefore, it remains necessary to develop more RNAi agents to inhibit or reduce MUC5AC expression for the treatment of mucus obstruction diseases. Summary of the Invention

[0008] This invention utilizes sequence design to screen for novel siRNAs with novel sequences capable of inhibiting MUC5AC mRNA activity, thereby blocking airway hypersecretion and providing an effective means for long-term treatment of mucus obstructive airway diseases. Chemical modification and ligand conjugation of the siRNA further enhance its inhibitory activity.

[0009] In a first aspect, this disclosure provides a double-stranded RNA (dsRNA) for inhibiting MUC5AC gene expression, said dsRNA comprising a sense strand and an antisense strand forming a double-stranded region, wherein the length of each of the sense strand and the antisense strand is independently 15-30 nucleotides, and said antisense strand comprises SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 9 2, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 16 0, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.In some implementations, the justice chain includes SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 9 1, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

[0010] In some implementations, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 56, 58, 60, 62, 64, 66, 68, 70, 74, 76, 78, 80, 82, 86, 92, 96, 98, 100, 102, 106, 110, 112, 116, 128, 130, 134, 152, 156, 158, 160, 162, 164, 166, 168, 170, 174, 176, 178, 180, 182, 184, 186 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278. In some embodiments, the positive strand comprises SEQ ID NO. 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238. NO:3,5,7,9,11,15,17,19,21,23,25,27,29,31,33,35,37,39,41,43,45,55,57,59,61,63,65,67,69,73,75,77,79,81,85,91 ,95,97,99,101,105,109,111,115,127,129,133,151,155,157,159,161,163,165,167,169,173,175,177,179,181,183,185, A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

[0011] In some implementations, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 62, 64, 66, 68, 74, 80, 82, 86, 102, 128, 156, 158, 162, 166, 168, 170, 176, 178, 182, 184, 186, 188, 190, 192, 194 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in SEQ ID NO: 196, 198, 204, 206, 208, 212, 214, 218, 220, 228, 230, 234, 236, 238, 240, 242, 244, 250, 252, 254, 256, 260, 264, 266, 268, 270, 272, 274, 278. In some embodiments, the positive strand comprises SEQ ID NO: 196, 198, 204, 206, 208, 212, 214, 218, 220, 228, 230, 234, 236, 238, 240, 242, 244, 250, 252, 254, 256, 260, 264, 266, 268, 270, 272, 274, 278. NO:3, 5, 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 61, 63, 65, 67, 73, 79, 8 1, 85, 101, 127, 155, 157, 161, 165, 167, 169, 175, 177, 181, 183, 185, 187, 189, 191, 193, A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 195, 197, 203, 205, 207, 211, 213, 217, 219, 227, 229, 233, 235, 237, 239, 241, 243, 249, 251, 253, 255, 259, 263, 265, 267, 269, 271, 273, 277.

[0012] In some embodiments, the antisense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 66, 86, 102, 128, 144, 156, 184, 192, 278. In some embodiments, the sense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 65, 85, 101, 127, 143, 155, 183, 191, 277.

[0013] In some implementations, the dsRNA is siRNA.

[0014] In some implementations, the lengths of the sense strand and the antisense strand are each independently 15-27 nucleotides, preferably 18-25 nucleotides, and more preferably 19-23 nucleotides.

[0015] In some implementations, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 9 6, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 16 6, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 2 A nucleotide sequence of at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleotides of any one of the nucleotide sequences shown in 36, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.Preferably, the antisense chain comprises SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88. 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 15 6, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220 The nucleotide sequence shown in any one of the following: 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.

[0016] In some implementations, the justice chain includes SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 9 3, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 1 63, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 2 A nucleotide sequence of at least 16, at least 17, or at least 18 consecutive nucleotides of any one of the nucleotide sequences shown in 31, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, and 277.Preferably, the justice chain comprises SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87. 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219 The nucleotide sequence shown is any one of the following: 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

[0017] In some embodiments, the dsRNA comprises any of the following pairs of paired sense and antisense sequences:

[0018] (1) Justice chain: SEQ ID NO:3, Antisense chain: SEQ ID NO:4;

[0019] (2) Justice chain: SEQ ID NO:5, Antisense chain: SEQ ID NO:6;

[0020] (3) Justice chain: SEQ ID NO:7, Antisense chain: SEQ ID NO:8;

[0021] (4) Justice chain: SEQ ID NO:9, Antisense chain: SEQ ID NO:10;

[0022] (5) Justice chain: SEQ ID NO:11, Antisense chain: SEQ ID NO:12;

[0023] (6) Justice chain: SEQ ID NO:13, Antisense chain: SEQ ID NO:14;

[0024] (7) Justice chain: SEQ ID NO:15, Antisense chain: SEQ ID NO:16;

[0025] (8) Justice Chain: SEQ ID NO:17, Antisense Chain: SEQ ID NO:18;

[0026] (9) Justice Chain: SEQ ID NO:19, Antisense Chain: SEQ ID NO:20;

[0027] (10) Justice chain: SEQ ID NO:21, Antisense chain: SEQ ID NO:22;

[0028] (11) Justice chain: SEQ ID NO:23, Antisense chain: SEQ ID NO:24;

[0029] (12) Justice chain: SEQ ID NO:25, Antisense chain: SEQ ID NO:26;

[0030] (13) Justice chain: SEQ ID NO:27, Antisense chain: SEQ ID NO:28;

[0031] (14) Justice chain: SEQ ID NO:29, Antisense chain: SEQ ID NO:30;

[0032] (15) Justice chain: SEQ ID NO:31, Antisense chain: SEQ ID NO:32;

[0033] (16) Justice chain: SEQ ID NO:33, Antisense chain: SEQ ID NO:34;

[0034] (17) Justice chain: SEQ ID NO:35, Antisense chain: SEQ ID NO:36;

[0035] (18) Justice chain: SEQ ID NO:37, Antisense chain: SEQ ID NO:38;

[0036] (19) Justice Chain: SEQ ID NO:39, Antisense Chain: SEQ ID NO:40;

[0037] (20) Justice chain: SEQ ID NO:41, Antisense chain: SEQ ID NO:42;

[0038] (21) Justice chain: SEQ ID NO:43, Antisense chain: SEQ ID NO:44;

[0039] (22) Justice chain: SEQ ID NO:45, Antisense chain: SEQ ID NO:46;

[0040] (23) Justice chain: SEQ ID NO:47, Antisense chain: SEQ ID NO:48;

[0041] (24) Justice chain: SEQ ID NO:49, Antisense chain: SEQ ID NO:50;

[0042] (25) Justice chain: SEQ ID NO:51, Antisense chain: SEQ ID NO:52;

[0043] (26) Justice chain: SEQ ID NO:53, Antisense chain: SEQ ID NO:54;

[0044] (27) Justice chain: SEQ ID NO:55, Antisense chain: SEQ ID NO:56;

[0045] (28) Justice chain: SEQ ID NO:57, Antisense chain: SEQ ID NO:58;

[0046] (29) Justice chain: SEQ ID NO:59, Antisense chain: SEQ ID NO:60;

[0047] (30) Justice chain: SEQ ID NO:61, Antisense chain: SEQ ID NO:62;

[0048] (31) Justice chain: SEQ ID NO:63, Antisense chain: SEQ ID NO:64;

[0049] (32) Justice chain: SEQ ID NO:65, Antisense chain: SEQ ID NO:66;

[0050] (33) Justice chain: SEQ ID NO:67, Antisense chain: SEQ ID NO:68;

[0051] (34) Justice chain: SEQ ID NO:69, Antisense chain: SEQ ID NO:70;

[0052] (35) Justice chain: SEQ ID NO:71, Antisense chain: SEQ ID NO:72;

[0053] (36) Justice chain: SEQ ID NO:73, Antisense chain: SEQ ID NO:74;

[0054] (37) Justice chain: SEQ ID NO:75, Antisense chain: SEQ ID NO:76;

[0055] (38) Justice chain: SEQ ID NO:77, Antisense chain: SEQ ID NO:78;

[0056] (39) Justice chain: SEQ ID NO:79, Antisense chain: SEQ ID NO:80;

[0057] (40) Justice chain: SEQ ID NO:81, Antisense chain: SEQ ID NO:82;

[0058] (41) Justice chain: SEQ ID NO:83, Antisense chain: SEQ ID NO:84;

[0059] (42) Justice chain: SEQ ID NO:85, Antisense chain: SEQ ID NO:86;

[0060] (43) Justice chain: SEQ ID NO:87, Antisense chain: SEQ ID NO:88;

[0061] (44) Justice chain: SEQ ID NO:89, Antisense chain: SEQ ID NO:90;

[0062] (45) Justice chain: SEQ ID NO:91, Antisense chain: SEQ ID NO:92;

[0063] (46) Justice chain: SEQ ID NO:93, Antisense chain: SEQ ID NO:94;

[0064] (47) Justice chain: SEQ ID NO:95, Antisense chain: SEQ ID NO:96;

[0065] (48) Justice chain: SEQ ID NO:97, Antisense chain: SEQ ID NO:98;

[0066] (49) Justice Chain: SEQ ID NO:99, Antisense Chain: SEQ ID NO:100;

[0067] (50) Justice chain: SEQ ID NO:101, Antisense chain: SEQ ID NO:102;

[0068] (51) Justice chain: SEQ ID NO:103, Antisense chain: SEQ ID NO:104;

[0069] (52) Justice chain: SEQ ID NO:105, Antisense chain: SEQ ID NO:106;

[0070] (53) Justice chain: SEQ ID NO:107, Antisense chain: SEQ ID NO:108;

[0071] (54) Justice chain: SEQ ID NO:109, Antisense chain: SEQ ID NO:110;

[0072] (55) Justice chain: SEQ ID NO:111, Antisense chain: SEQ ID NO:112;

[0073] (56) Justice chain: SEQ ID NO:113, Antisense chain: SEQ ID NO:114;

[0074] (57) Justice chain: SEQ ID NO:115, Antisense chain: SEQ ID NO:116;

[0075] (58) Justice chain: SEQ ID NO:117, Antisense chain: SEQ ID NO:118;

[0076] (59) Justice chain: SEQ ID NO:119, Antisense chain: SEQ ID NO:120;

[0077] (60) Justice chain: SEQ ID NO:121, Antisense chain: SEQ ID NO:122;

[0078] (61) Justice chain: SEQ ID NO:123, Antisense chain: SEQ ID NO:124;

[0079] (62) Justice chain: SEQ ID NO:125, Antisense chain: SEQ ID NO:126;

[0080] (63) Justice chain: SEQ ID NO:127, Antisense chain: SEQ ID NO:128;

[0081] (64) Justice chain: SEQ ID NO:129, Antisense chain: SEQ ID NO:130;

[0082] (65) Justice chain: SEQ ID NO:131, Antisense chain: SEQ ID NO:132;

[0083] (66) Justice chain: SEQ ID NO:133, Antisense chain: SEQ ID NO:134;

[0084] (67) Justice chain: SEQ ID NO:135, Antisense chain: SEQ ID NO:136;

[0085] (68) Justice chain: SEQ ID NO:137, Antisense chain: SEQ ID NO:138;

[0086] (69) Justice chain: SEQ ID NO:139, Antisense chain: SEQ ID NO:140;

[0087] (70) Justice chain: SEQ ID NO:141, Antisense chain: SEQ ID NO:142;

[0088] (71) Justice chain: SEQ ID NO:143, Antisense chain: SEQ ID NO:144;

[0089] (72) Justice chain: SEQ ID NO:145, Antisense chain: SEQ ID NO:146;

[0090] (73) Justice chain: SEQ ID NO:147, Antisense chain: SEQ ID NO:148;

[0091] (74) Justice chain: SEQ ID NO:149, Antisense chain: SEQ ID NO:150;

[0092] (75) Justice chain: SEQ ID NO:151, Antisense chain: SEQ ID NO:152;

[0093] (76) Justice chain: SEQ ID NO:153, Antisense chain: SEQ ID NO:154;

[0094] (77) Justice chain: SEQ ID NO:155, Antisense chain: SEQ ID NO:156;

[0095] (78) Justice chain: SEQ ID NO:157, Antisense chain: SEQ ID NO:158;

[0096] (79) Justice chain: SEQ ID NO:159, Antisense chain: SEQ ID NO:160;

[0097] (80) Justice chain: SEQ ID NO:161, Antisense chain: SEQ ID NO:162;

[0098] (81) Justice chain: SEQ ID NO:163, Antisense chain: SEQ ID NO:164;

[0099] (82) Justice chain: SEQ ID NO:165, Antisense chain: SEQ ID NO:166;

[0100] (83) Justice chain: SEQ ID NO:167, Antisense chain: SEQ ID NO:168;

[0101] (84) Justice chain: SEQ ID NO:169, Antisense chain: SEQ ID NO:170;

[0102] (85) Justice chain: SEQ ID NO:171, Antisense chain: SEQ ID NO:172;

[0103] (86) Justice chain: SEQ ID NO:173, Antisense chain: SEQ ID NO:174;

[0104] (87) Justice chain: SEQ ID NO:175, Antisense chain: SEQ ID NO:176;

[0105] (88) Justice Chain: SEQ ID NO:177, Antisense Chain: SEQ ID NO:178;

[0106] (89) Justice chain: SEQ ID NO:179, Antisense chain: SEQ ID NO:180;

[0107] (90) Justice chain: SEQ ID NO:181, Antisense chain: SEQ ID NO:182;

[0108] (91) Justice chain: SEQ ID NO:183, Antisense chain: SEQ ID NO:184;

[0109] (92) Justice chain: SEQ ID NO:185, Antisense chain: SEQ ID NO:186;

[0110] (93) Justice chain: SEQ ID NO:187, Antisense chain: SEQ ID NO:188;

[0111] (94) Justice Chain: SEQ ID NO:189, Antisense Chain: SEQ ID NO:190;

[0112] (95) Justice chain: SEQ ID NO:191, Antisense chain: SEQ ID NO:192;

[0113] (96) Justice Chain: SEQ ID NO:193, Antisense Chain: SEQ ID NO:194;

[0114] (97) Justice chain: SEQ ID NO:195, Antisense chain: SEQ ID NO:196;

[0115] (98) Justice Chain: SEQ ID NO:197, Antisense Chain: SEQ ID NO:198;

[0116] (99) Justice Chain: SEQ ID NO:199, Antisense Chain: SEQ ID NO:200;

[0117] (100) Justice chain: SEQ ID NO:201, Antisense chain: SEQ ID NO:202;

[0118] (101) Justice chain: SEQ ID NO:203, Antisense chain: SEQ ID NO:204;

[0119] (102) Justice chain: SEQ ID NO:205, Antisense chain: SEQ ID NO:206;

[0120] (103) Justice chain: SEQ ID NO:207, Antisense chain: SEQ ID NO:208;

[0121] (104) Justice Chain: SEQ ID NO:209, Antisense Chain: SEQ ID NO:210;

[0122] (105) Justice chain: SEQ ID NO:211, Antisense chain: SEQ ID NO:212;

[0123] (106) Justice chain: SEQ ID NO:213, Antisense chain: SEQ ID NO:214;

[0124] (107) Justice chain: SEQ ID NO:215, Antisense chain: SEQ ID NO:216;

[0125] (108) Justice Chain: SEQ ID NO:217, Antisense Chain: SEQ ID NO:218;

[0126] (109) Justice Chain: SEQ ID NO:219, Antisense Chain: SEQ ID NO:220;

[0127] (110) Justice chain: SEQ ID NO:221, Antisense chain: SEQ ID NO:222;

[0128] (111) Justice chain: SEQ ID NO:223, Antisense chain: SEQ ID NO:224;

[0129] (112) Justice chain: SEQ ID NO:225, Antisense chain: SEQ ID NO:226;

[0130] (113) Justice chain: SEQ ID NO:227, Antisense chain: SEQ ID NO:228;

[0131] (114) Justice chain: SEQ ID NO:229, Antisense chain: SEQ ID NO:230;

[0132] (115) Justice chain: SEQ ID NO:231, Antisense chain: SEQ ID NO:232;

[0133] (116) Justice chain: SEQ ID NO:233, Antisense chain: SEQ ID NO:234;

[0134] (117) Justice chain: SEQ ID NO:235, Antisense chain: SEQ ID NO:236;

[0135] (118) Justice chain: SEQ ID NO:237, Antisense chain: SEQ ID NO:238;

[0136] (119) Justice chain: SEQ ID NO:239, Antisense chain: SEQ ID NO:240;

[0137] (120) Justice chain: SEQ ID NO:241, Antisense chain: SEQ ID NO:242;

[0138] (121) Justice chain: SEQ ID NO:243, Antisense chain: SEQ ID NO:244;

[0139] (122) Justice chain: SEQ ID NO:245, Antisense chain: SEQ ID NO:246;

[0140] (123) Justice chain: SEQ ID NO:247, Antisense chain: SEQ ID NO:248;

[0141] (124) Justice chain: SEQ ID NO:249, Antisense chain: SEQ ID NO:250;

[0142] (125) Justice chain: SEQ ID NO:251, Antisense chain: SEQ ID NO:252;

[0143] (126) Justice chain: SEQ ID NO:253, Antisense chain: SEQ ID NO:254;

[0144] (127) Justice chain: SEQ ID NO:255, Antisense chain: SEQ ID NO:256;

[0145] (128) Justice chain: SEQ ID NO:257, Antisense chain: SEQ ID NO:258;

[0146] (129) Justice chain: SEQ ID NO:259, Antisense chain: SEQ ID NO:260;

[0147] (130) Justice chain: SEQ ID NO:261, Antisense chain: SEQ ID NO:262;

[0148] (131) Justice chain: SEQ ID NO:263, Antisense chain: SEQ ID NO:264;

[0149] (132) Justice chain: SEQ ID NO:265, Antisense chain: SEQ ID NO:266;

[0150] (133) Justice chain: SEQ ID NO:267, Antisense chain: SEQ ID NO:268;

[0151] (134) Justice chain: SEQ ID NO:269, Antisense chain: SEQ ID NO:270;

[0152] (135) Justice chain: SEQ ID NO:271, Antisense chain: SEQ ID NO:272;

[0153] (136) Justice chain: SEQ ID NO:273, Antisense chain: SEQ ID NO:274;

[0154] (137) Justice chain: SEQ ID NO:275, Antisense chain: SEQ ID NO:276;

[0155] (138) Justice chain: SEQ ID NO:277, Antisense chain: SEQ ID NO:278.

[0156] In some embodiments, at least one nucleotide of the sense strand and / or the antisense strand is a modified nucleotide.

[0157] In some embodiments, substantially all nucleotides of the sense strand and substantially all nucleotides of the antisense strand are modified nucleotides, or all nucleotides of the sense strand and all nucleotides of the antisense strand are modified nucleotides.

[0158] In some embodiments, the sense strand and the antisense strand each independently comprise one or more nucleotide modifications selected from the group consisting of: 2'-O-methyl modification, 2'-O-methoxyethyl modification, 2'-fluoro modification, DNA modification, glycerol nucleic acid (GNA) modification, unlocking nucleic acid (UNA) modification, vinylphosphonate modification (e.g., 5'-VP modification), inverse non-base deoxyribonucleotide (invAb) modification, and phosphate thioester nucleotide interbonding modification. In a preferred embodiment, the sense strand and the antisense strand each independently comprise one or more nucleotide modifications selected from the group consisting of: 2'-O-methyl modification, 2'-fluoro modification, DNA modification, phosphate thioester nucleotide interbonding modification, 5'-VP modification, and invAb modification.

[0159] In some embodiments, the antisense strand has a length of 23 nucleotides, and the sense strand has a length of 21 nucleotides.

[0160] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 5, 11, 13, 15, 17, 19, 21, 22, and 23, counted from the 5' end; preferably, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 13, 15, 17, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 2, 14, and 16.

[0161] In some implementations, the antisense chain has:

[0162] (i) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 12, 14, and 16; or

[0163] (ii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 7, 8, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 9, 12, 14, and 16; or

[0164] (iii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 5, 6, 7, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 4, 12, 14, and 16; or

[0165] (iv) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 10, 14, and 16; or

[0166] (v) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20; or

[0167] (vi) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 19, 20, 21, 22, and 23, and the nucleotides with 2'-fluoro modification at positions 2, 6, 14, 16, and 18; or

[0168] (vii) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the nucleotides with 2'-fluoro modification at positions 2, 8, 9, 14, and 16; or

[0169] (viii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 7, 14, and 16; or

[0170] (ix) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 14, and 16; or

[0171] (x) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the nucleotides with 2'-fluorine modification at positions 2, 7, 12, 14, and 16;

[0172] and / or

[0173] (xi) Starting from the 5' end, count the phosphate ester nucleotide bonds located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23;

[0174] and / or

[0175] (xii) VP modification at the 5' end.

[0176] In some embodiments, the positive strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at position 9; preferably, the positive strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 9 and 11; more preferably, the positive strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 16, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 7, 9, and 11.

[0177] In some implementations, the justice chain has:

[0178] (i) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, and 11; or

[0179] (ii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 9, 11, and 13; or

[0180] (iii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 15; or

[0181] (iv) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21; the 2'-fluoro modified nucleotides at positions 7, 9, and 10; and the deoxyribonucleotides at position 11; or

[0182] (v) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 13; or

[0183] (vi) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 16; or

[0184] (vii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 17; or

[0185] (viii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 12; or

[0186] (ix) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and the 2'-fluoro modified nucleotides at positions 8, 9, 10, 11; or

[0187] (x) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 15, 16, 17, 18, 19, 20, 21, and the nucleotides with 2'-fluorine modification at positions 9, 11, 14;

[0188] and / or

[0189] (xi) Starting from the 5' end, count the internucleotide bonds of phosphate thioesters located between nucleotide positions 1 and 2 and between 20 and 21;

[0190] and / or

[0191] (xii) is modified by invAb at the 3' end.

[0192] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 12, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, and 11, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0193] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 12, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 9, 11, and 13, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0194] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 7, 8, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 9, 12, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 9, 11, and 13, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0195] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 5, 6, 7, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 4, 12, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, and 11, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0196] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 10, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 15, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0197] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 10, 14, and 16; and phosphate thiophosphates located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Ester-nucleotide inter-bonding; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end; 2'-fluoro modified nucleotides at positions 7, 9, and 10; and a deoxyribonucleotide at position 11, as well as thiophosphate ester nucleotide inter-bondings between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0198] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides located at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20; and phosphate thiophosphates located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Ester-nucleotide inter-bonding; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end; 2'-fluoro modified nucleotides at positions 7, 9, and 10; and a deoxyribonucleotide at position 11, as well as thiophosphate ester nucleotide inter-bondings between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0199] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 6, 14, 16, and 18; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 13, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0200] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 8, 9, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 16, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0201] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 8, 9, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 17, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0202] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides located at positions 2, 7, 14, and 16; and phosphate thioesters located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Nucleotide-to-nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end; 2'-fluorinated modified nucleotides at positions 7, 9, 10, 11, and 12; and invAb modification at the 3' end; and phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0203] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 8, 9, 10, and 11, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0204] In some embodiments, the antisense strand is 23 nucleotides long and has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23 (counting from the 5' end), 2'-fluoro modified nucleotides at positions 2, 7, 12, 14, and 16, and a phosphate thioester core at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23 (counting from the 5' end). The nucleotides are bonded together, and there is a VP modification at the 5' end; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 9, 11, and 14, and phosphate thioester nucleotides bonded together between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0205] In some implementations, the antisense chain comprises any antisense chain sequence selected from those shown in Table 8.

[0206] In some implementations, the justice chain comprises any of the justice chain sequences shown in Table 8.

[0207] In some embodiments, the dsRNA comprises a pair of sense and antisense sequences selected from any of the pairs shown in Table 8.

[0208] In some embodiments, the dsRNA comprises any of the following pairs of paired sense and antisense sequences:

[0209] (1) Justice chain: SEQ ID NO:293, Antisense chain: SEQ ID NO:294;

[0210] (2) Justice chain: SEQ ID NO:399, Antisense chain: SEQ ID NO:400;

[0211] (3) Justice chain: SEQ ID NO:479, Antisense chain: SEQ ID NO:480;

[0212] (4) Justice chain: SEQ ID NO:527, Antisense chain: SEQ ID NO:528;

[0213] (5) Justice chain: SEQ ID NO:533, Antisense chain: SEQ ID NO:534;

[0214] (6) Justice chain: SEQ ID NO:595, Antisense chain: SEQ ID NO:596;

[0215] (7) Justice chain: SEQ ID NO:597, Antisense chain: SEQ ID NO:598;

[0216] (8) Justice chain: SEQ ID NO:607, Antisense chain: SEQ ID NO:608;

[0217] (9) Justice chain: SEQ ID NO:617, Antisense chain: SEQ ID NO:618;

[0218] (10) Justice chain: SEQ ID NO:627, Antisense chain: SEQ ID NO:628;

[0219] (11) Justice chain: SEQ ID NO:687, Antisense chain: SEQ ID NO:688;

[0220] (12) Justice chain: SEQ ID NO:693, Antisense chain: SEQ ID NO:694;

[0221] (13) Justice chain: SEQ ID NO:697, Antisense chain: SEQ ID NO:698;

[0222] (14) Justice chain: SEQ ID NO:739, Antisense chain: SEQ ID NO:740;

[0223] (15) Justice chain: SEQ ID NO:787, Antisense chain: SEQ ID NO:788;

[0224] (16) Justice chain: SEQ ID NO:797, Antisense chain: SEQ ID NO:798;

[0225] (17) Justice chain: SEQ ID NO:801, Antisense chain: SEQ ID NO:802.

[0226] In some implementations, the dsRNA is further conjugated with a target ligand, preferably with the positive strand of the target ligand, and more preferably with the 5' end of the positive strand of the target ligand.

[0227] In some embodiments, the targeting ligand is an integrin targeting ligand, preferably an αvβ6 integrin targeting ligand.

[0228] In some implementations, the targeting ligand has the following structure:

[0229] in This indicates the position where the dsRNA is attached to the positive strand via a phosphate group or a thiophosphate group.

[0230] In some embodiments, the dsRNA comprises a pair of paired sense and antisense strand sequences selected from the following, and the 5' end of the sense strand is conjugated to the ligand Tri-SM6.1 via a phosphate thioester or phosphodiester bond:

[0231] (1) Justice chain: SEQ ID NO:399, Antisense chain: SEQ ID NO:400;

[0232] (2) Justice chain: SEQ ID NO:479, Antisense chain: SEQ ID NO:480;

[0233] (3) Justice chain: SEQ ID NO:533, Antisense chain: SEQ ID NO:534;

[0234] (4) Justice chain: SEQ ID NO:597, Antisense chain: SEQ ID NO:598;

[0235] (5) Justice chain: SEQ ID NO:607, Antisense chain: SEQ ID NO:608;

[0236] (6) Justice chain: SEQ ID NO:617, Antisense chain: SEQ ID NO:618;

[0237] (7) Justice chain: SEQ ID NO:627, Antisense chain: SEQ ID NO:628.

[0238] In a second aspect, this disclosure provides a vector comprising a nucleotide sequence encoding the dsRNA described in the first aspect of this disclosure.

[0239] In a third aspect, this disclosure provides cells comprising the dsRNA described in the first aspect of this disclosure or the vector described in the second aspect of this disclosure.

[0240] In a fourth aspect, this disclosure provides pharmaceutical compositions comprising the dsRNA described in the first aspect of this disclosure, the vector described in the second aspect of this disclosure, or the cell described in the third aspect of this disclosure, and optionally a pharmaceutically acceptable carrier or excipient.

[0241] In a fifth aspect, this disclosure provides a kit comprising the dsRNA described in the first aspect of this disclosure, the vector described in the second aspect of this disclosure, or the cells described in the third aspect of this disclosure.

[0242] In a sixth aspect, this disclosure provides a method for inhibiting the expression of the MUC5AC gene in cells, the method comprising contacting the cells with the dsRNA described in the first aspect of this disclosure or the pharmaceutical composition described in the fourth aspect of this disclosure.

[0243] In a seventh aspect, this disclosure provides a method for inhibiting the expression of the MUC5AC gene in the cells of a subject, the method comprising administering to the subject the dsRNA described in the first aspect of this disclosure, the vector described in the second aspect of this disclosure, the cells described in the third aspect of this disclosure, or the pharmaceutical composition described in the fourth aspect of this disclosure.

[0244] In an eighth aspect, this disclosure provides a method for treating a disease or condition in a subject that benefits from reduced MUC5AC gene expression, the method comprising administering to the subject an effective amount of the dsRNA described in the first aspect of this disclosure, the vector described in the second aspect of this disclosure, the cell described in the third aspect of this disclosure, or the pharmaceutical composition described in the fourth aspect of this disclosure.

[0245] In some implementations, the disease or condition that benefits from reduced MUC5AC gene expression is a disease mediated or associated with MUC5AC expression.

[0246] In some implementations, the diseases mediated or associated with the MUC5AC expression are respiratory diseases.

[0247] In some embodiments, the diseases mediated or associated with the MUC5AC expression are mucus obstructive airway diseases, preferably selected from asthma, COPD, and bronchiectasis. Detailed Implementation

[0248] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0249] It should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terminology used in the embodiments of the present invention is for describing specific embodiments and not for limiting the scope of protection of the present invention.

[0250] In this application specification and claims, unless otherwise expressly stated herein, the singular forms “an,” “a,” and “the” include the plural forms.

[0251] Unless otherwise stated or defined, the terms “comprising,” “including,” and “containing” should be understood to mean that the said element or step or group of elements or steps are included, but do not exclude any other element or step or group of elements or steps.

[0252] The term "and / or" as used herein includes any one of two or more related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected using at least two conjunctions selected from "and / or," it should be understood that, in this application, the technical solution undoubtedly includes solutions connected by "logical AND," and also undoubtedly includes solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, AND / OR, B, AND / OR, C, AND / OR, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0253] When a numerical range is given in the implementation scheme, it should be understood that, unless otherwise stated in the present invention, the two endpoints of each numerical range and any value between the two endpoints may be selected.

[0254] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. Apart from the specific methods, apparatus, and materials used in the embodiments, based on the prior art mastery of one of ordinary skill in the art and the description of this invention, any prior art methods, apparatus, and materials similar to or equivalent to those described, used, or made by those of ordinary skill in the art in implementing this invention may be used to implement this invention, and all such methods, apparatus, and materials shall fall within the protection scope of this invention. The embodiments of this invention are described in more detail below.

[0255] I. Definition

[0256] In this paper, a “double-stranded region” refers to a region containing two antiparallel and complementary or substantially complementary nucleic acid strands.

[0257] As used herein, the term "double-stranded RNA" or "dsRNA" refers to a ribonucleic acid molecule or ribonucleic acid molecule complex that contains a double-stranded region as defined above. The two parts forming the double-stranded region can be two distinct parts of a larger RNA molecule, or they can be separate RNA molecules.

[0258] When the two parts are separate RNA molecules, the dsRNA in this article is referred to as small interfering RNA or short interfering RNA, or simply siRNA.

[0259] When two parts are two distinct portions of a larger molecule, i.e., when the 3' end of one part is linked to the 5' end of the other part to form a double-stranded region by one or more uninterrupted nucleotides, the uninterrupted nucleotide used for the linkage is called a "hairpin loop." When the two parts are covalently linked to form a double-stranded region by a method other than a hairpin loop, the linkage structure is called a "linkosome." After such dsRNA is introduced into the cell, it is cleaved into siRNA by an intracellular endonuclease called Dicer.

[0260] As used herein, the term "siRNA" refers to a class of double-stranded RNA molecules comprising a sense strand and an antisense strand that can mediate the silencing of a target RNA (e.g., mRNA, such as a transcript of a gene encoding a protein) that is complementary or substantially complementary to the antisense strand. siRNAs are typically double-stranded, consisting of an antisense strand complementary to the target RNA and a sense strand complementary or substantially complementary to that antisense strand. Typically, the target RNA is an endogenous gene or a pathogen gene. Additionally, RNAs other than mRNA (e.g., tRNA) and viral RNAs can also be targeted.

[0261] As used herein, the term “antisense strand” refers to a strand in dsRNA (especially siRNA) that contains regions that are fully or substantially complementary to the target sequence.

[0262] As used herein, the term "complementary region" refers to a region on the antisense strand that is fully or substantially complementary to the target mRNA sequence. In cases where the complementary region is not fully complementary to the target sequence, mismatches can occur within the molecule or at the ends. Typically, the most tolerant mismatches are located in the end regions, for example, within 5, 4, 3, 2, or 1 nucleotides at the 5' and / or 3' ends. The portion of the antisense strand most sensitive to mismatches is called the "seed region." For example, in a siRNA containing a 19-nt strand, the 19th position (from 5' to 3') can tolerate some mismatches.

[0263] As used herein, the term "complementarity" refers to the ability of a first polynucleotide to hybridize with a second polynucleotide under certain conditions, such as stringent conditions. For example, stringent conditions may include 400 mM NaCl, 40 mM PIPES at pH 6.4, and 1 mM EDTA at 50 or 70 °C for 12–16 hours.

[0264] As used herein, in order to satisfy the above requirements regarding their hybridization ability, "complementary" sequences may also include base pairs formed entirely from non-Watson-Crick base pairs and / or from non-natural and modified nucleotides. Such non-Watson-Crick base pairs include, but are not limited to, G:U swing base pairings or Hoogstein base pairs.

[0265] As used herein, a polynucleotide that is “at least partially complementary” or “substantially complementary” to mRNA refers to a polynucleotide that is substantially complementary to a continuous portion of the mRNA of interest (e.g., the mRNA encoding MUC5AC). For example, a polynucleotide is at least partially complementary to the mRNA encoding MUC5AC if the sequence is substantially complementary to an uninterrupted portion of the mRNA encoding MUC5AC.

[0266] As used herein, the terms “complementary,” “fully complementary,” and “substantially complementary” can be used relative to the base pairing between the sense and antisense strands of dsRNA, particularly siRNA, or between the antisense strand of dsRNA, particularly siRNA, and the target sequence.

[0267] As used herein, the term "sense chain" refers to a chain of siRNA that includes regions substantially complementary to the regions defined herein as antisense chains.

[0268] "Nucleoside" is a compound composed of two substances: a purine or pyrimidine base and ribose or deoxyribose. "Nucleotide" is a compound composed of three substances: a purine or pyrimidine base, ribose or deoxyribose, and phosphate. "Oligonucleotide" refers to nucleic acid molecules (RNA or DNA) with a length of less than 100, 200, 300, or 400 nucleotides.

[0269] A "base" is the basic building block for the synthesis of nucleosides, nucleotides, and nucleic acids. Its constituent elements include nitrogen, hence it is also called a "nitrogenous base." In this article, unless otherwise specified, the capital letters A, U, T, G, and C represent the base composition of nucleotides, namely adenine, uracil, thymine, guanine, and cytosine, respectively.

[0270] As used herein, the term "nucleotide overhang" refers to at least one unpaired nucleotide that protrudes from the double-stranded region of siRNA. A nucleotide overhang exists, for example, when the 3' end of one strand of siRNA extends beyond the 5' end of the other strand (or vice versa). siRNA may contain overhangs having at least one nucleotide, at least two nucleotides, at least three nucleotides, at least four nucleotides, or at least five or more nucleotides. A nucleotide overhang may contain or consist of nucleotides / modified nucleotides (including deoxynucleotides / nucleosides). One or more overhangs may be located on the sense strand or antisense strand, or a combination thereof. An overhang having one or more nucleotides may be present at the 5' end, 3' end, or both ends of the antisense or sense strand of siRNA.

[0271] "Flat-ended" or "blunt-ended" means that there are no unpaired nucleotides at the ends of the double-stranded siRNA, i.e., no nucleotide overhangs. "Flat-ended siRNA" is a double-stranded siRNA that is double-stranded throughout its entire length, meaning that there are no nucleotide overhangs at either end of the molecule.

[0272] As used herein, the term "modified nucleotide" refers to a nucleotide other than a ribonucleotide (2'-hydroxynucleotide), which can be obtained by modification methods known in the art. The phrase "substantially all nucleotides are modified" as used herein means that the majority, but not all, nucleotides of the dsRNA (especially siRNA) of this application are modified, and may include no more than 5, 4, 3, 2, or 1 unmodified nucleotides.

[0273] "Modified nucleotides" may include, but are not limited to, 2'-O-alkyl modified nucleotides (e.g., 2'-O-methyl modified nucleotides, 2'-methoxyethyl modified nucleotides), 2'-fluoro modified nucleotides, 2'-deoxy modified nucleotides (e.g., deoxyribonucleotides), inosine ribonucleotides, abase-free nucleotides, reverse abase-free deoxyribonucleotides, nucleotides containing a thiophosphate group, thiophosphate ester nucleotide bonds, vinylphosphonate modified nucleotides, locked nucleotides, unlocked nucleotides, 2'-amino modified nucleotides, and 2'-C-alkyl modified nucleotides. Nucleotides, 2'-O-allyl modified nucleotides, morpholinonucleotides, aminophosphates, nucleotides containing non-natural bases, terminal nucleotides linked to cholesterol derivatives or dodecanoic acid di ...

[0274] As used herein, the term "2'-O-methyl modified nucleotide" refers to a nucleotide in which the hydroxyl group at the 2' position of the ribosyl group is replaced by a methoxy group. "2'-Fluoromodified nucleotide" refers to a nucleotide in which the hydroxyl group at the 2' position of the ribosyl group is replaced by a fluorine group; it is also called "2'-deoxy-2'-fluorine modified nucleotide." "Modification of internucleotide thiophosphate bonds" refers to a modification in which two adjacent nucleotides are linked by a thiophosphate ester. "Nucleotide containing a thiophosphate group" refers to a nucleotide in which one or more oxygen atoms on the phosphate group are replaced by sulfur atoms. "5'-VP modification" refers to 5'-(E)-vinylphosphonate modification, which is a modification that replaces the phosphate group at the 5' end of the antisense strand with an (E)-vinylphosphonate structure. "DNA modification," also known as "deoxyribonucleotide modification," refers to the replacement of ribonucleotides with their corresponding deoxyribonucleotides; that is, U is replaced with dT; A with dA; C with dC; and G with dG. "invAb modification" refers to reverse abase-free deoxyribonucleotide modification. invAb is a nucleotide analog whose structural feature is the inversion of the stereochemical configuration of the deoxyribose ring, resulting in the interchange of the 5' and 3' ends of the sugar ring. This modification is usually introduced into the 3' or 5' end of the siRNA double strand to interfere with the recognition and binding of nucleases, thereby enhancing the stability of the siRNA molecule.

[0275] As used herein, a “targeting ligand” refers to a chemical moiety conjugated to dsRNA (especially siRNA) that can alter the distribution, targeting, or lifespan of dsRNA (especially siRNA). For example, a targeting ligand provides enhanced affinity for selected targets (e.g., molecules, cells or cell types, compartments, e.g., cellular or organ compartments, tissues, organs, or regions of the body) compared to siRNA without a targeting ligand.

[0276] As used herein, “αvβ6 integrin targeting ligand” is a compound with affinity for integrin αvβ6 that can be used as a ligand to facilitate the targeting and delivery of its linked dsRNA to desired cells and / or tissues (i.e., cells expressing integrin αvβ6, such as epithelial cells).

[0277] As used herein, the term “pharmaceutical acceptable” means that the carrier or excipient is compatible with the other components of the composition and is not substantially toxic to the recipient, and / or that such carrier or excipient is approved or may be used in a pharmaceutical composition administered to a subject.

[0278] As used herein, the term “inhibition” is used interchangeably with “reduction,” “silence,” “downregulation,” and other similar terms, and includes any level of inhibition.

[0279] The phrase "inhibit MUC5AC gene expression" refers to the inhibition of the expression of any MUC5AC gene, as well as variants or mutants of the MUC5AC gene. Therefore, the MUC5AC gene can be a wild-type MUC5AC gene, a mutant MUC5AC gene, or a transgenic MUC5AC gene in the case of genetically manipulated cells, cell groups, or organisms.

[0280] "Inhibition of MUC5AC gene expression" includes inhibition of any level of the MUC5AC gene, such as at least partial inhibition of MUC5AC gene expression. MUC5AC gene expression can be assessed based on the level or changes in the level of any variable associated with MUC5AC gene expression, such as the mRNA level encoding MUC5AC or the protein level of MUC5AC. This level can be assessed in a single cell or in a group of cells, including, for example, samples derived from a subject.

[0281] Inhibition can be assessed by a decrease in the absolute or relative level of one or more variables associated with MUC5AC gene expression compared to a control level. The control level can be any type of control level utilized in the art, such as baseline levels before administration or levels determined from similar untreated or controlled (e.g., buffer-only control or inert agent control) subjects, cells, or samples.

[0282] As used herein, the terms “treatment,” “therapy,” “treatment,” etc., refer to the administration of a drug or the conduct of a procedure for the purpose of achieving an effect. These effects may be preventative in the complete or partial prevention of a disease or its symptoms, and / or therapeutic in the partial or complete cure of a disease and / or its symptoms. As used herein, “treatment” can include treating a disease or condition (e.g., an inflammatory disease) in mammals, particularly humans, and includes: (a) preventing the onset of a disease or its symptoms in subjects who may be susceptible to the disease (e.g., including diseases that may be related to or caused by the primary disease) but have not yet been diagnosed with the disease; (b) suppressing the disease, i.e., halting its progression; and (c) alleviating the disease, i.e., causing its regression. Treatment can refer to any indicator of success in the treatment, improvement, or prevention of cancer, including any objective or subjective parameter such as a reduction in symptoms; remission; elimination of disease symptoms or making the disease condition more tolerable for the patient; slowing the rate of deterioration or decline; or weakening the final stage of deterioration. Treatment or improvement of symptoms is based on one or more objective or subjective parameters; including the results of a physician’s examination. Therefore, the term "treatment" includes the administration of the antibodies, compositions, or conjugates disclosed herein to prevent or delay, alleviate, or prevent or inhibit the development of symptoms or conditions associated with a disease (e.g., an inflammatory disease). The term "therapeutic effect" refers to the reduction, elimination, or prevention of disease, disease symptoms, or disease side effects in a subject.

[0283] The term "effective amount" as used in this invention refers to an amount sufficient to treat a disease when administered to a subject for the purpose of treating that disease.

[0284] As used herein, the term “subject” refers to any mammalian subject for diagnosis, treatment, or therapy. For therapeutic purposes, “mammal” means any animal classified as a mammal, including humans, livestock, and laboratory animals, zoo animals, sporting animals, or pet animals, such as dogs, horses, cats, cattle, sheep, goats, pigs, mice, rats, rabbits, guinea pigs, monkeys, etc.

[0285] II. dsRNA

[0286] This disclosure provides a double-stranded RNA (dsRNA) for suppressing MUC5AC gene expression, said dsRNA comprising a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand and the antisense strand are each independently 15-30 nucleotides in length. In some embodiments, the antisense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of any of the antisense strand sequences shown in Table 1. In some embodiments, the sense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of any of the sense strand sequences shown in Table 1.

[0287] In some implementations, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 9 2, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 16 0, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.In some implementations, the justice chain includes SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 9 1, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

[0288] In some implementations, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 56, 58, 60, 62, 64, 66, 68, 70, 74, 76, 78, 80, 82, 86, 92, 96, 98, 100, 102, 106, 110, 112, 116, 128, 130, 134, 152, 156, 158, 160, 162, 164, 166, 168, 170, 174, 176, 178, 180, 182, 184, 186 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278. In some embodiments, the positive strand comprises SEQ ID NO. 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238. NO:3,5,7,9,11,15,17,19,21,23,25,27,29,31,33,35,37,39,41,43,45,55,57,59,61,63,65,67,69,73,75,77,79,81,85,91 ,95,97,99,101,105,109,111,115,127,129,133,151,155,157,159,161,163,165,167,169,173,175,177,179,181,183,185, A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

[0289] In some implementations, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 62, 64, 66, 68, 74, 80, 82, 86, 102, 128, 156, 158, 162, 166, 168, 170, 176, 178, 182, 184, 186, 188, 190, 192, 194 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in SEQ ID NO: 196, 198, 204, 206, 208, 212, 214, 218, 220, 228, 230, 234, 236, 238, 240, 242, 244, 250, 252, 254, 256, 260, 264, 266, 268, 270, 272, 274, 278. In some embodiments, the positive strand comprises SEQ ID NO: 196, 198, 204, 206, 208, 212, 214, 218, 220, 228, 230, 234, 236, 238, 240, 242, 244, 250, 252, 254, 256, 260, 264, 266, 268, 270, 272, 274, 278. NO:3, 5, 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 61, 63, 65, 67, 73, 79, 8 1, 85, 101, 127, 155, 157, 161, 165, 167, 169, 175, 177, 181, 183, 185, 187, 189, 191, 193, A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 195, 197, 203, 205, 207, 211, 213, 217, 219, 227, 229, 233, 235, 237, 239, 241, 243, 249, 251, 253, 255, 259, 263, 265, 267, 269, 271, 273, 277.

[0290] In some embodiments, the antisense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 66, 86, 102, 128, 144, 156, 184, 192, 278. In some embodiments, the sense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 65, 85, 101, 127, 143, 155, 183, 191, 277.

[0291] In some embodiments, the length of the positive and negative strands is independently 15-27 nucleotides, preferably 18-25 nucleotides, more preferably 19-23 nucleotides, and even more preferably 21-23 nucleotides. In some embodiments, the length of the positive and negative strands is independently 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.

[0292] In some embodiments, the length of the double-stranded region is 15-25 nucleotide pairs, preferably 16-23 nucleotide pairs, and more preferably 18-21 nucleotide pairs.

[0293] In some implementations, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 9 8, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 24 A nucleotide sequence of at least 15, at least 16, at least 17, at least 18, at least 19, at least 20 or at least 21 consecutive nucleotides of any one of the nucleotide sequences shown in 0, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.In some embodiments, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236 The sequence of any one of the numbers 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278 differs by no more than 0, 1, 2, or 3 nucleotides and consists of at least 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides.In a preferred embodiment, the antisense chain comprises SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88. 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 15 6, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220 The nucleotide sequence shown in any one of the following: 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278. In a more preferred embodiment, the antisense chain includes 4, 6, 8, 10, 12, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 56, 58, 60, 62, 64, 66, 68, 70, 74, 76, 78, 80, 82, 86, 92, 96, 98, 100, 102, 106, 110, 112, 116, 128, 130, 134, 152, 156, 158, 160, 162, 164, 166, 168, 170, 174, 176, 178 The nucleotide sequence shown is any one of the following: 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.In a further preferred embodiment, the antisense chain includes SEQ ID NO: 4, 6, 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 62, 64, 66, 68, 74, 80, 82, 86, 102, 128, 156, 158, 162, 166, 168, 170, 176, 178, 182, 184, 186, 188, 190 The nucleotide sequence shown is any one of the following: 192, 194, 196, 198, 204, 206, 208, 212, 214, 218, 220, 228, 230, 234, 236, 238, 240, 242, 244, 250, 252, 254, 256, 260, 264, 266, 268, 270, 272, 274, 278.

[0294] In some implementations, the justice chain includes SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 9 7, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 1 69, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239 A nucleotide sequence of at least 15, at least 16, at least 17, at least 18, at least 19, at least 20 or at least 21 consecutive nucleotides of any of the nucleotide sequences shown in 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.In some implementations, the justice chain includes SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 1 67, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 2 The sequence shown in any one of the sequences 37, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, and 277 differs from the sequence by no more than 0, 1, 2, or 3 nucleotides in at least 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides.In a preferred embodiment, the justice chain comprises SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87. 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219 The nucleotide sequence shown in any one of SEQ ID NO. 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277. In a more preferred embodiment, the positive strand comprises ... NO:3,5,7,9,11,15,17,19,21,23,25,27,29,31,33,35,37,39,41,43,45,55,57,59,61,63,65,67,69,73,75,77,79,81,8 5, 91, 95, 97, 99, 101, 105, 109, 111, 115, 127, 129, 133, 151, 155, 157, 159, 161, 163, 165, 167, 169, 173, 175, 177, 179, 181, The nucleotide sequence shown is any one of the following: 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.In a further preferred embodiment, the justice chain comprises SEQ ID NO: 3, 5, 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 61, 63, 65, 67, 73, 79, 81, 85, 101, 127, 155, 157, 161, 165, 167, 169, 175, 177, 181, 183, 185, 187, 189 The nucleotide sequence shown is any one of the following: 191, 193, 195, 197, 203, 205, 207, 211, 213, 217, 219, 227, 229, 233, 235, 237, 239, 241, 243, 249, 251, 253, 255, 259, 263, 265, 267, 269, 271, 273, 277.

[0295] In some embodiments, the antisense strand comprises the nucleotide sequence shown in any one of SEQ ID NO: 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 66, 86, 102, 128, 144, 156, 184, 192, 278. In some embodiments, the sense strand comprises the nucleotide sequence shown in any one of SEQ ID NO: 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 65, 85, 101, 127, 143, 155, 183, 191, 277.

[0296] In some embodiments, the dsRNA is siRNA. In other embodiments, a hairpin loop is formed between the sense and antisense strands of the dsRNA of the present invention.

[0297] The dsRNA of the present invention includes dsRNA having a nucleotide overhang at one end (i.e., having one overhang and one blunt end) or having nucleotide overhangs at both ends. For example, the 5' end of the sense strand of the dsRNA includes an overhang of one or more nucleotides and the 3' end of the sense strand includes an overhang of one or more nucleotides. For example, the 5' end of the antisense strand of the dsRNA includes an overhang of one or more nucleotides and the 3' end of the antisense strand includes an overhang of one or more nucleotides. For example, the 5' end of the sense strand of the dsRNA includes an overhang of one or more nucleotides and the 5' end of the antisense strand includes an overhang of one or more nucleotides. For example, the 3' end of the sense strand of the dsRNA includes an overhang of one or more nucleotides and the 3' end of the antisense strand includes an overhang of one or more nucleotides. For example, the 5' end of the sense strand of the dsRNA includes an overhang of one or more nucleotides and the 3' end of the sense strand includes a blunt end. For example, the 3' end of the sense strand of dsRNA contains a protruding end with one or more nucleotides, and the 5' end of the sense strand contains a blunt end. Similarly, the 5' end of the antisense strand of dsRNA contains a protruding end with one or more nucleotides, and the 3' end of the antisense strand contains a blunt end.

[0298] In some embodiments, the 3' end of the antisense strand of the dsRNA of the present invention includes a protrusion having one or more nucleotides, and the 5' end of the antisense strand includes a blunt end. In some embodiments, the 3' end of the antisense strand of the dsRNA of the present invention includes a protrusion having one, two, three, or four nucleotides, and the 5' end of the antisense strand includes a blunt end. In some embodiments, the 3' end of the antisense strand of the dsRNA of the present invention includes a protrusion having two nucleotides, and the 5' end of the antisense strand includes a blunt end.

[0299] In some embodiments, the 3' end of the sense strand of the dsRNA of the present invention includes a protrusion having one or more nucleotides, and the 5' end of the sense strand includes a blunt end. In some embodiments, the 3' end of the sense strand of the dsRNA of the present invention includes a protrusion having one, two, three, or four nucleotides, and the 5' end of the sense strand includes a blunt end. In some embodiments, the 3' end of the sense strand of the dsRNA of the present invention includes a protrusion having two nucleotides, and the 5' end of the sense strand includes a blunt end.

[0300] In some embodiments, one or both of the sense strand and the antisense strand include a 3' overhang and / or a 5' overhang having at least one nucleotide. In some specific embodiments, one or both of the sense strand and the antisense strand include a 3' overhang and / or a 5' overhang having one nucleotide. In some specific embodiments, one or both of the sense strand and the antisense strand include a 3' overhang and / or a 5' overhang having two nucleotides. In some specific embodiments, one or both of the sense strand and the antisense strand include a 3' overhang and / or a 5' overhang having three nucleotides. In some specific embodiments, one or both of the sense strand and the antisense strand include a 3' overhang and / or a 5' overhang having four nucleotides.

[0301] In some specific embodiments, the antisense strand includes a 3' overhang and / or a 5' overhang having one nucleotide. In some embodiments, the antisense strand includes a 3' overhang and / or a 5' overhang having two nucleotides. In some embodiments, the antisense strand includes a 3' overhang and / or a 5' overhang having three nucleotides. In some embodiments, the antisense strand includes a 3' overhang and / or a 5' overhang having four nucleotides. In some preferred embodiments, the antisense strand has a 3' overhang and / or a 5' overhang having at least two nucleotides. Preferably, the antisense strand includes a 3' overhang and / or a 5' overhang having two nucleotides.

[0302] In some specific embodiments, the positive strand includes a 3' overhang and / or a 5' overhang of 1 nucleotide. In some embodiments, the positive strand includes a 3' overhang and / or a 5' overhang of 2 nucleotides. In some embodiments, the positive strand includes a 3' overhang and / or a 5' overhang of 3 nucleotides. In some embodiments, the positive strand includes a 3' overhang and / or a 5' overhang of 4 nucleotides. In some preferred embodiments, the positive strand has a 3' overhang and / or a 5' overhang of at least 2 nucleotides. Preferably, the positive strand includes a 3' overhang and / or a 5' overhang of 2 nucleotides.

[0303] In some implementations, the justice chain and the antisense chain are of the same length.

[0304] In some implementations, the full length of the justice chain is complementary to the full length of the antisense chain, forming a double chain, i.e., having flat ends.

[0305] In other embodiments, the sense strand and the antisense strand are of the same length, and a portion of the sense strand is complementary to a portion of the antisense strand, i.e., both the sense strand and the antisense strand have a 3' or 5' overhang. In some embodiments, the sense strand and the antisense strand are of different lengths. In a preferred embodiment, the 3' or 5' end of the antisense strand has an overhang of at least one nucleotide, more preferably two or three nucleotides.

[0306] In a preferred embodiment, the antisense strand includes a 3' overhang of 2 nucleotides. In other preferred embodiments, the sense strand includes a 3' overhang of 2 nucleotides.

[0307] In some implementations, the bistranded regions formed by the sense and antisense strands are completely complementary. In other implementations, the bistranded regions formed by the sense and antisense strands are substantially complementary, and may contain one, two, three, four, or five non-complementary sites.

[0308] In some embodiments, the dsRNA comprises any pair of paired sense and antisense sequences as shown in Table 1. In some embodiments, the dsRNA comprises any of the following pairs of paired sense and antisense sequences:

[0309] (1) Justice chain: SEQ ID NO:3, Antisense chain: SEQ ID NO:4;

[0310] (2) Justice chain: SEQ ID NO:5, Antisense chain: SEQ ID NO:6;

[0311] (3) Justice chain: SEQ ID NO:7, Antisense chain: SEQ ID NO:8;

[0312] (4) Justice chain: SEQ ID NO:9, Antisense chain: SEQ ID NO:10;

[0313] (5) Justice chain: SEQ ID NO:11, Antisense chain: SEQ ID NO:12;

[0314] (6) Justice chain: SEQ ID NO:13, Antisense chain: SEQ ID NO:14;

[0315] (7) Justice chain: SEQ ID NO:15, Antisense chain: SEQ ID NO:16;

[0316] (8) Justice Chain: SEQ ID NO:17, Antisense Chain: SEQ ID NO:18;

[0317] (9) Justice Chain: SEQ ID NO:19, Antisense Chain: SEQ ID NO:20;

[0318] (10) Justice chain: SEQ ID NO:21, Antisense chain: SEQ ID NO:22;

[0319] (11) Justice chain: SEQ ID NO:23, Antisense chain: SEQ ID NO:24;

[0320] (12) Justice chain: SEQ ID NO:25, Antisense chain: SEQ ID NO:26;

[0321] (13) Justice chain: SEQ ID NO:27, Antisense chain: SEQ ID NO:28;

[0322] (14) Justice chain: SEQ ID NO:29, Antisense chain: SEQ ID NO:30;

[0323] (15) Justice chain: SEQ ID NO:31, Antisense chain: SEQ ID NO:32;

[0324] (16) Justice chain: SEQ ID NO:33, Antisense chain: SEQ ID NO:34;

[0325] (17) Justice chain: SEQ ID NO:35, Antisense chain: SEQ ID NO:36;

[0326] (18) Justice chain: SEQ ID NO:37, Antisense chain: SEQ ID NO:38;

[0327] (19) Justice Chain: SEQ ID NO:39, Antisense Chain: SEQ ID NO:40;

[0328] (20) Justice chain: SEQ ID NO:41, Antisense chain: SEQ ID NO:42;

[0329] (21) Justice chain: SEQ ID NO:43, Antisense chain: SEQ ID NO:44;

[0330] (22) Justice chain: SEQ ID NO:45, Antisense chain: SEQ ID NO:46;

[0331] (23) Justice chain: SEQ ID NO:47, Antisense chain: SEQ ID NO:48;

[0332] (24) Justice chain: SEQ ID NO:49, Antisense chain: SEQ ID NO:50;

[0333] (25) Justice chain: SEQ ID NO:51, Antisense chain: SEQ ID NO:52;

[0334] (26) Justice chain: SEQ ID NO:53, Antisense chain: SEQ ID NO:54;

[0335] (27) Justice chain: SEQ ID NO:55, Antisense chain: SEQ ID NO:56;

[0336] (28) Justice chain: SEQ ID NO:57, Antisense chain: SEQ ID NO:58;

[0337] (29) Justice chain: SEQ ID NO:59, Antisense chain: SEQ ID NO:60;

[0338] (30) Justice chain: SEQ ID NO:61, Antisense chain: SEQ ID NO:62;

[0339] (31) Justice chain: SEQ ID NO:63, Antisense chain: SEQ ID NO:64;

[0340] (32) Justice chain: SEQ ID NO:65, Antisense chain: SEQ ID NO:66;

[0341] (33) Justice chain: SEQ ID NO:67, Antisense chain: SEQ ID NO:68;

[0342] (34) Justice chain: SEQ ID NO:69, Antisense chain: SEQ ID NO:70;

[0343] (35) Justice chain: SEQ ID NO:71, Antisense chain: SEQ ID NO:72;

[0344] (36) Justice chain: SEQ ID NO:73, Antisense chain: SEQ ID NO:74;

[0345] (37) Justice chain: SEQ ID NO:75, Antisense chain: SEQ ID NO:76;

[0346] (38) Justice chain: SEQ ID NO:77, Antisense chain: SEQ ID NO:78;

[0347] (39) Justice chain: SEQ ID NO:79, Antisense chain: SEQ ID NO:80;

[0348] (40) Justice chain: SEQ ID NO:81, Antisense chain: SEQ ID NO:82;

[0349] (41) Justice chain: SEQ ID NO:83, Antisense chain: SEQ ID NO:84;

[0350] (42) Justice chain: SEQ ID NO:85, Antisense chain: SEQ ID NO:86;

[0351] (43) Justice chain: SEQ ID NO:87, Antisense chain: SEQ ID NO:88;

[0352] (44) Justice chain: SEQ ID NO:89, Antisense chain: SEQ ID NO:90;

[0353] (45) Justice chain: SEQ ID NO:91, Antisense chain: SEQ ID NO:92;

[0354] (46) Justice chain: SEQ ID NO:93, Antisense chain: SEQ ID NO:94;

[0355] (47) Justice chain: SEQ ID NO:95, Antisense chain: SEQ ID NO:96;

[0356] (48) Justice chain: SEQ ID NO:97, Antisense chain: SEQ ID NO:98;

[0357] (49) Justice Chain: SEQ ID NO:99, Antisense Chain: SEQ ID NO:100;

[0358] (50) Justice chain: SEQ ID NO:101, Antisense chain: SEQ ID NO:102;

[0359] (51) Justice chain: SEQ ID NO:103, Antisense chain: SEQ ID NO:104;

[0360] (52) Justice chain: SEQ ID NO:105, Antisense chain: SEQ ID NO:106;

[0361] (53) Justice chain: SEQ ID NO:107, Antisense chain: SEQ ID NO:108;

[0362] (54) Justice chain: SEQ ID NO:109, Antisense chain: SEQ ID NO:110;

[0363] (55) Justice chain: SEQ ID NO:111, Antisense chain: SEQ ID NO:112;

[0364] (56) Justice chain: SEQ ID NO:113, Antisense chain: SEQ ID NO:114;

[0365] (57) Justice chain: SEQ ID NO:115, Antisense chain: SEQ ID NO:116;

[0366] (58) Justice chain: SEQ ID NO:117, Antisense chain: SEQ ID NO:118;

[0367] (59) Justice chain: SEQ ID NO:119, Antisense chain: SEQ ID NO:120;

[0368] (60) Justice chain: SEQ ID NO:121, Antisense chain: SEQ ID NO:122;

[0369] (61) Justice chain: SEQ ID NO:123, Antisense chain: SEQ ID NO:124;

[0370] (62) Justice chain: SEQ ID NO:125, Antisense chain: SEQ ID NO:126;

[0371] (63) Justice chain: SEQ ID NO:127, Antisense chain: SEQ ID NO:128;

[0372] (64) Justice chain: SEQ ID NO:129, Antisense chain: SEQ ID NO:130;

[0373] (65) Justice chain: SEQ ID NO:131, Antisense chain: SEQ ID NO:132;

[0374] (66) Justice chain: SEQ ID NO:133, Antisense chain: SEQ ID NO:134;

[0375] (67) Justice chain: SEQ ID NO:135, Antisense chain: SEQ ID NO:136;

[0376] (68) Justice chain: SEQ ID NO:137, Antisense chain: SEQ ID NO:138;

[0377] (69) Justice chain: SEQ ID NO:139, Antisense chain: SEQ ID NO:140;

[0378] (70) Justice chain: SEQ ID NO:141, Antisense chain: SEQ ID NO:142;

[0379] (71) Justice chain: SEQ ID NO:143, Antisense chain: SEQ ID NO:144;

[0380] (72) Justice chain: SEQ ID NO:145, Antisense chain: SEQ ID NO:146;

[0381] (73) Justice chain: SEQ ID NO:147, Antisense chain: SEQ ID NO:148;

[0382] (74) Justice chain: SEQ ID NO:149, Antisense chain: SEQ ID NO:150;

[0383] (75) Justice chain: SEQ ID NO:151, Antisense chain: SEQ ID NO:152;

[0384] (76) Justice chain: SEQ ID NO:153, Antisense chain: SEQ ID NO:154;

[0385] (77) Justice chain: SEQ ID NO:155, Antisense chain: SEQ ID NO:156;

[0386] (78) Justice chain: SEQ ID NO:157, Antisense chain: SEQ ID NO:158;

[0387] (79) Justice chain: SEQ ID NO:159, Antisense chain: SEQ ID NO:160;

[0388] (80) Justice chain: SEQ ID NO:161, Antisense chain: SEQ ID NO:162;

[0389] (81) Justice chain: SEQ ID NO:163, Antisense chain: SEQ ID NO:164;

[0390] (82) Justice chain: SEQ ID NO:165, Antisense chain: SEQ ID NO:166;

[0391] (83) Justice chain: SEQ ID NO:167, Antisense chain: SEQ ID NO:168;

[0392] (84) Justice chain: SEQ ID NO:169, Antisense chain: SEQ ID NO:170;

[0393] (85) Justice chain: SEQ ID NO:171, Antisense chain: SEQ ID NO:172;

[0394] (86) Justice chain: SEQ ID NO:173, Antisense chain: SEQ ID NO:174;

[0395] (87) Justice chain: SEQ ID NO:175, Antisense chain: SEQ ID NO:176;

[0396] (88) Justice Chain: SEQ ID NO:177, Antisense Chain: SEQ ID NO:178;

[0397] (89) Justice chain: SEQ ID NO:179, Antisense chain: SEQ ID NO:180;

[0398] (90) Justice chain: SEQ ID NO:181, Antisense chain: SEQ ID NO:182;

[0399] (91) Justice chain: SEQ ID NO:183, Antisense chain: SEQ ID NO:184;

[0400] (92) Justice chain: SEQ ID NO:185, Antisense chain: SEQ ID NO:186;

[0401] (93) Justice chain: SEQ ID NO:187, Antisense chain: SEQ ID NO:188;

[0402] (94) Justice Chain: SEQ ID NO:189, Antisense Chain: SEQ ID NO:190;

[0403] (95) Justice chain: SEQ ID NO:191, Antisense chain: SEQ ID NO:192;

[0404] (96) Justice Chain: SEQ ID NO:193, Antisense Chain: SEQ ID NO:194;

[0405] (97) Justice chain: SEQ ID NO:195, Antisense chain: SEQ ID NO:196;

[0406] (98) Justice Chain: SEQ ID NO:197, Antisense Chain: SEQ ID NO:198;

[0407] (99) Justice Chain: SEQ ID NO:199, Antisense Chain: SEQ ID NO:200;

[0408] (100) Justice chain: SEQ ID NO:201, Antisense chain: SEQ ID NO:202;

[0409] (101) Justice chain: SEQ ID NO:203, Antisense chain: SEQ ID NO:204;

[0410] (102) Justice chain: SEQ ID NO:205, Antisense chain: SEQ ID NO:206;

[0411] (103) Justice chain: SEQ ID NO:207, Antisense chain: SEQ ID NO:208;

[0412] (104) Justice Chain: SEQ ID NO:209, Antisense Chain: SEQ ID NO:210;

[0413] (105) Justice chain: SEQ ID NO:211, Antisense chain: SEQ ID NO:212;

[0414] (106) Justice chain: SEQ ID NO:213, Antisense chain: SEQ ID NO:214;

[0415] (107) Justice chain: SEQ ID NO:215, Antisense chain: SEQ ID NO:216;

[0416] (108) Justice Chain: SEQ ID NO:217, Antisense Chain: SEQ ID NO:218;

[0417] (109) Justice Chain: SEQ ID NO:219, Antisense Chain: SEQ ID NO:220;

[0418] (110) Justice chain: SEQ ID NO:221, Antisense chain: SEQ ID NO:222;

[0419] (111) Justice chain: SEQ ID NO:223, Antisense chain: SEQ ID NO:224;

[0420] (112) Justice chain: SEQ ID NO:225, Antisense chain: SEQ ID NO:226;

[0421] (113) Justice chain: SEQ ID NO:227, Antisense chain: SEQ ID NO:228;

[0422] (114) Justice chain: SEQ ID NO:229, Antisense chain: SEQ ID NO:230;

[0423] (115) Justice chain: SEQ ID NO:231, Antisense chain: SEQ ID NO:232;

[0424] (116) Justice chain: SEQ ID NO:233, Antisense chain: SEQ ID NO:234;

[0425] (117) Justice chain: SEQ ID NO:235, Antisense chain: SEQ ID NO:236;

[0426] (118) Justice chain: SEQ ID NO:237, Antisense chain: SEQ ID NO:238;

[0427] (119) Justice chain: SEQ ID NO:239, Antisense chain: SEQ ID NO:240;

[0428] (120) Justice chain: SEQ ID NO:241, Antisense chain: SEQ ID NO:242;

[0429] (121) Justice chain: SEQ ID NO:243, Antisense chain: SEQ ID NO:244;

[0430] (122) Justice chain: SEQ ID NO:245, Antisense chain: SEQ ID NO:246;

[0431] (123) Justice chain: SEQ ID NO:247, Antisense chain: SEQ ID NO:248;

[0432] (124) Justice chain: SEQ ID NO:249, Antisense chain: SEQ ID NO:250;

[0433] (125) Justice chain: SEQ ID NO:251, Antisense chain: SEQ ID NO:252;

[0434] (126) Justice chain: SEQ ID NO:253, Antisense chain: SEQ ID NO:254;

[0435] (127) Justice chain: SEQ ID NO:255, Antisense chain: SEQ ID NO:256;

[0436] (128) Justice chain: SEQ ID NO:257, Antisense chain: SEQ ID NO:258;

[0437] (129) Justice chain: SEQ ID NO:259, Antisense chain: SEQ ID NO:260;

[0438] (130) Justice chain: SEQ ID NO:261, Antisense chain: SEQ ID NO:262;

[0439] (131) Justice chain: SEQ ID NO:263, Antisense chain: SEQ ID NO:264;

[0440] (132) Justice chain: SEQ ID NO:265, Antisense chain: SEQ ID NO:266;

[0441] (133) Justice chain: SEQ ID NO:267, Antisense chain: SEQ ID NO:268;

[0442] (134) Justice chain: SEQ ID NO:269, Antisense chain: SEQ ID NO:270;

[0443] (135) Justice chain: SEQ ID NO:271, Antisense chain: SEQ ID NO:272;

[0444] (136) Justice chain: SEQ ID NO:273, Antisense chain: SEQ ID NO:274;

[0445] (137) Justice chain: SEQ ID NO:275, Antisense chain: SEQ ID NO:276;

[0446] (138) Justice chain: SEQ ID NO:277, Antisense chain: SEQ ID NO:278.

[0447] In the dsRNA (especially siRNA) of the present invention, all nucleotide groups may be unmodified or contain at least one modified nucleotide group, and the modification may be on nucleotides at any position.

[0448] III. Modified nucleotides

[0449] The dsRNA (especially siRNA) of the present invention may be modified, for example, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, or at least 23 nucleotides of the sense strand and / or the antisense strand are modified nucleotides. Essentially all nucleotides of the dsRNA (especially siRNA) of the present invention are modified. For example, essentially all nucleotides of the sense strand are modified nucleotides, or essentially all nucleotides of the antisense strand are modified nucleotides, or essentially all nucleotides of both the sense and antisense strands are modified nucleotides. In some embodiments, at least 80%, at least 85%, at least 90%, at least 92%, or at least 95% of the nucleotides in the sense strand are modified nucleotides, and / or at least 80%, at least 85%, at least 90%, at least 92%, or at least 95% of the nucleotides in the antisense strand are modified nucleotides. In other embodiments, all nucleotides of the dsRNA (especially siRNA) of the present invention are modified nucleotides. For example, all nucleotides in the sense strand are modified nucleotides, or all nucleotides in the antisense strand are modified nucleotides, or all nucleotides in both the sense and antisense strands are modified nucleotides.

[0450] The nucleotide modification described in this invention can be any chemical modification known in the art. Chemical modifications can be classified into three different types based on their site: 1) phosphate modification; 2) ribose modification; and 3) base modification. Therefore, the nucleotide modification described in this invention can be a modification on the phosphate group, ribose group, and / or base group of the nucleotide.

[0451] Various methods for modifying nucleotides of siRNA have been developed in this field. For example, Alnylam, Arrowhead Pharmaceuticals, Silence, Dicerna, and others have developed chemical modification schemes for nucleotides to improve the drug-likeness of siRNA.

[0452] For example, the evolution of Alnylam modification schemes mainly went through the following processes. First was the STC modification scheme, which employed the following modifications: 1) ribose modification, including 2'-deoxy-2'-fluorine (2'-F) and 2'-OMe; and 2) phosphate modification, using PS (phosphothiophosphate) modification. STC modification is a universal modification scheme that can increase siRNA stability and affinity for mRNA. Subsequently, Alnylam's next-generation siRNA modification scheme, ESC, emerged. ESC modification has four more PS modifications than STC modification, and another significant change is the reduction in the number of 2'-F modified nucleotides. This reduction in 2'-F is mainly because excessive use of 2'-F modification may increase siRNA toxicity. Compared to the STC modification scheme, ESC significantly improves the efficacy and half-life of siRNA. The ESC modification scheme gives siRNA drugs better PK and PD, while reducing the dosage of siRNA drugs. Alnylam subsequently optimized the ESC modification scheme, designing the advanced ESC modification scheme. Advanced ESC further reduced 2'-F modification and optimized the ratio and modification sites of 2'-F and 2'-OMe. Advanced ESC modification further improved the stability of siRNA without affecting its effectiveness. Alnylam then further developed the ESC+ modification scheme, which adds GNA modification to the seed sequence portion of the siRNA antisense strand based on ESC modification. This modification primarily addresses the off-target effects and hepatotoxicity of miRNA-like siRNAs.

[0453] In some specific embodiments, the sense strand and the antisense strand each independently comprise one or more nucleotide modifications selected from the group consisting of: 2'-O-alkyl modified nucleotides (e.g., 2'-O-methyl modified nucleotides, 2'-methoxyethyl modified nucleotides), 2'-fluoro modified nucleotides, 2'-deoxy modified nucleotides (e.g., deoxyribonucleotides), inosine ribonucleotides, abase-free nucleotides, inverse abase-free deoxyribonucleotides (invAb), nucleotides containing a thiophosphate group, thiophosphate ester nucleotide bonds, vinylphosphonate modified nucleotides (e.g., 5'-VP modified nucleotides), locked nucleotides, unlocked nucleotides. Nucleotides, 2'-amino modified nucleotides, 2'-C-alkyl modified nucleotides, 2'-O-allyl modified nucleotides, morpholinonucleotides, aminophosphates, nucleotides containing non-natural bases, terminal nucleotides linked to cholesterol derivatives or dodecanoic acid di ...

[0454] In some embodiments, the sense strand and the antisense strand each independently comprise one or more nucleotide modifications selected from the group consisting of: 2'-O-methyl modification, 2'-O-methoxyethyl modification, 2'-fluoro modification, DNA modification, glycerol nucleotide (GNA) modification, unlocking nucleotide (UNA) modification, vinylphosphonate modification, reverse abase-free deoxyribonucleotide modification, and phosphate-thioester nucleotide-to-nucleotide bonding modification. In some embodiments, the sense strand and the antisense strand each independently comprise one or more modified nucleotides selected from the group consisting of: 2'-O-methyl modified nucleotides, 2'-O-methoxyethyl modified nucleotides, 2'-fluoro modified nucleotides, deoxyribonucleotides, glycerol nucleotides, unlocking nucleotides, vinylphosphonate modified nucleotides, reverse abase-free deoxyribonucleotides, and nucleotides containing a phosphate-thioester group. In a preferred embodiment, the sense strand and the antisense strand each independently comprise one or more nucleotide modifications selected from the group consisting of: 2'-O-methyl modification, 2'-fluoro modification, DNA modification, modification of phosphate thioester nucleotide inter-linking, 5'-VP modification, and invAb modification.

[0455] In some embodiments, the positive and / or negative strands comprise at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 2'-O-methyl modified nucleotides. In some preferred embodiments, the positive and / or negative strands comprise at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 2'-fluoro modified nucleotides. In some embodiments, the 3' and / or 5' ends of the positive and / or negative strands comprise 1-5 phosphate thioester nucleotide bonds, for example, 1-4 phosphate thioester nucleotide bonds, preferably 2 or 4 phosphate thioester nucleotide bonds. In some embodiments, the sense strand and / or the antisense strand have phosphate-thiophosphate nucleotide bonds located between nucleotide positions 20 and 21 or 22 and 23, counted from the 5' end. In some embodiments, the sense strand and / or the antisense strand have phosphate-thiophosphate nucleotide bonds located between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3, counted from the 5' end. In some embodiments, the sense strand and / or the antisense strand have phosphate-thiophosphate nucleotide bonds located between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end. In some embodiments, the sense strand and / or the antisense strand have phosphate-thiophosphate nucleotide bonds located between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23, counted from the 5' end. In some embodiments, the sense strand and / or the antisense strand has a deoxyribonucleotide located at nucleotide position 11, counted from the 5' end. In some implementations, the sense chain and / or the antisense chain has an invAb modifier located at the 5' or 3' end.

[0456] In some embodiments, the antisense strand has a length of 23 nucleotides, and the sense strand has a length of 21 nucleotides.

[0457] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 5, 11, 13, 15, 17, 19, 21, 22, and 23, counted from the 5' end. In a preferred embodiment, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 13, 15, 17, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 2, 14, and 16.

[0458] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 6, 8, 9, 12, 14, and 16, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0459] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 7, 8, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 9, 12, 14, and 16, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0460] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 5, 6, 7, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 4, 12, 14, and 16, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0461] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 6, 8, 9, 10, 14, and 16, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0462] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0463] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 6, 14, 16, and 18, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0464] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 8, 9, 14, and 16, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0465] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 7, 14, and 16, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0466] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 6, 8, 9, 14, and 16, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end, and optionally a VP modification at the 5' end.

[0467] In some embodiments, the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluorinated modified nucleotides at positions 2, 7, 12, 14, and 16; and thiophosphate nucleotide inter-bondings between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end; and optionally, a VP modification at the 5' end.

[0468] In some embodiments, the positive strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at position 9. In a preferred embodiment, the positive strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 9 and 11. In a more preferred embodiment, the positive strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 16, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 7, 9, and 11.

[0469] In some embodiments, the positive chain has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 7, 9, and 11, as well as phosphate thioester nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end, and optionally an invAb modification at the 3' end.

[0470] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 9, 11, and 13, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0471] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 15, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0472] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, 2'-fluoro modified nucleotides at positions 7, 9, and 10, and deoxyribonucleotides at position 11, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, starting from the 5' end, and optionally, invAb modification at the 3' end.

[0473] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 13, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0474] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 16, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0475] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 17, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0476] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 12, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0477] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 8, 9, 10, and 11, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0478] In some embodiments, starting from the 5' end, 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 9, 11, and 14, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, and optionally, invAb modification at the 3' end, are counted.

[0479] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 12, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, and 11, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0480] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 12, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 9, 11, and 13, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0481] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 7, 8, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 9, 12, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 9, 11, and 13, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0482] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 5, 6, 7, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 4, 12, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and 2'-fluoro modified nucleotides at positions 7, 9, and 11, and thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0483] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 10, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 15, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0484] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 10, 14, and 16; and phosphate thiophosphates located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Ester-nucleotide inter-bonding; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end; 2'-fluoro modified nucleotides at positions 7, 9, and 10; and a deoxyribonucleotide at position 11, as well as thiophosphate ester nucleotide inter-bondings between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0485] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides located at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20; and phosphate thiophosphates located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Ester-nucleotide inter-bonding; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end; 2'-fluoro modified nucleotides at positions 7, 9, and 10; and a deoxyribonucleotide at position 11, as well as thiophosphate ester nucleotide inter-bondings between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0486] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 6, 14, 16, and 18; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 13, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0487] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 8, 9, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 16, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0488] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluorinated modified nucleotides located at positions 2, 8, 9, 14, and 16; and nucleotides located between positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 17, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0489] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides located at positions 2, 7, 14, and 16; and phosphate thioesters located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Nucleotide-to-nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end; 2'-fluorinated modified nucleotides at positions 7, 9, 10, 11, and 12; and invAb modification at the 3' end; and phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end.

[0490] In some embodiments, the antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides located at positions 1, 3, 4, 5, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; and 2'-fluoro modified nucleotides located at positions 2, 6, 8, 9, 14, and 16; and nucleotides located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. The positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 8, 9, 10, and 11, as well as thiophosphate nucleotide bonds between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0491] In some embodiments, the antisense strand is 23 nucleotides long and has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23 (counting from the 5' end), 2'-fluoro modified nucleotides at positions 2, 7, 12, 14, and 16, and a phosphate thioester core at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23 (counting from the 5' end). The nucleotides are bonded together, and there is a VP modification at the 5' end; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 9, 11, and 14, and phosphate thioester nucleotides bonded together between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

[0492] In some embodiments, the antisense strand comprises any antisense strand sequence selected from those shown in Table 8. In some embodiments, the sense strand comprises any sense strand sequence selected from those shown in Table 8. In some embodiments, the dsRNA comprises any pair of paired sense and antisense strand sequences selected from those shown in Table 8.

[0493] In some embodiments, the dsRNA comprises any of the following pairs of paired sense and antisense sequences:

[0494] (1) Justice chain: SEQ ID NO:293, Antisense chain: SEQ ID NO:294;

[0495] (2) Justice chain: SEQ ID NO:399, Antisense chain: SEQ ID NO:400;

[0496] (3) Justice chain: SEQ ID NO:479, Antisense chain: SEQ ID NO:480;

[0497] (4) Justice chain: SEQ ID NO:527, Antisense chain: SEQ ID NO:528;

[0498] (5) Justice chain: SEQ ID NO:533, Antisense chain: SEQ ID NO:534;

[0499] (6) Justice chain: SEQ ID NO:595, Antisense chain: SEQ ID NO:596;

[0500] (7) Justice chain: SEQ ID NO:597, Antisense chain: SEQ ID NO:598;

[0501] (8) Justice chain: SEQ ID NO:607, Antisense chain: SEQ ID NO:608;

[0502] (9) Justice chain: SEQ ID NO:617, Antisense chain: SEQ ID NO:618;

[0503] (10) Justice chain: SEQ ID NO:627, Antisense chain: SEQ ID NO:628;

[0504] (11) Justice chain: SEQ ID NO:687, Antisense chain: SEQ ID NO:688;

[0505] (12) Justice chain: SEQ ID NO:693, Antisense chain: SEQ ID NO:694;

[0506] (13) Justice chain: SEQ ID NO:697, Antisense chain: SEQ ID NO:698;

[0507] (14) Justice chain: SEQ ID NO:739, Antisense chain: SEQ ID NO:740;

[0508] (15) Justice chain: SEQ ID NO:787, Antisense chain: SEQ ID NO:788;

[0509] (16) Justice chain: SEQ ID NO:797, Antisense chain: SEQ ID NO:798;

[0510] (17) Justice chain: SEQ ID NO:801, Antisense chain: SEQ ID NO:802.

[0511] IV. Targeted Ligands

[0512] The dsRNA (especially siRNA) of the present invention can be delivered to target cells or tissues using any oligonucleotide delivery technique known in the art. For example, the dsRNA of the present invention can be delivered to cells or tissues by conjugating a targeting ligand. Therefore, the dsRNA of the present invention can be conjugated to a targeting ligand.

[0513] In some embodiments, the targeting ligand has affinity for a cellular receptor expressed on epithelial cells. In some embodiments, the targeting ligand comprises an integrin targeting ligand. Integrins are a family of transmembrane receptors that promote cell-extracellular matrix (ECM) adhesion. For example, integrin α-v-β-6 (αvβ6) is an epithelial-specific integrin that is not expressed in healthy adult epithelial cells but is upregulated during embryogenesis, tissue repair, and tumorigenesis. αvβ6 is known to be significantly upregulated in various epithelial cancers, especially at the tumor invasive margin, and is therefore closely associated with tumor invasion. Integrin αvβ6 is also known to be highly upregulated in damaged lung epithelium. In some embodiments, the integrin targeting ligand is an αvβ6 integrin targeting ligand. The application of the αvβ6 integrin targeting ligand promotes specific targeting of cells with αvβ6 on their surface, and the binding of the integrin targeting ligand can facilitate the entry of its linked dsRNA into cells (e.g., epithelial cells, including lung and kidney epithelial cells). The siRNA-αvβ6 integrin targeting ligand conjugate obtained by conjugating the siRNA of the present invention with the αvβ6 integrin targeting ligand is selectively internalized by epithelial cells (e.g., lung epithelial cells) via receptor-mediated endocytosis or other mechanisms.

[0514] The targeting ligand can be monomeric or monovalent (e.g., having a single targeting moiety) or polymeric or multivalent (e.g., having multiple targeting moiety). Using methods known in the art, the targeting ligand can be attached to the 3' and / or 5' ends of the sense or antisense strand of the dsRNA of the present invention. In some embodiments, the sense strand of the dsRNA of the present invention is conjugated to the targeting ligand. In some embodiments, the 3' end of the sense strand is conjugated to the targeting ligand. In other embodiments, the 5' end of the sense strand is conjugated to the targeting ligand.

[0515] The targeting ligand can be any αvβ6 integrin targeting ligand known in the art, such as RGD peptides, RGD peptide mimics, lipids, steroids, vitamins, sugars, proteins, peptides, polyamines, etc. Examples of targeting ligands including αvβ6 integrin targeting ligands can be found, for example, in International Patent Application Publication Nos. WO 2018 / 085415 and WO 2019 / 089765, the disclosures of which are incorporated herein by reference in their entirety.

[0516] In some implementations, the targeting ligand has the following structure:

[0517] in This indicates the position where the dsRNA is attached to the positive strand via a phosphate group or a thiophosphate group.

[0518] In some embodiments, the dsRNA comprises any pair of paired sense and antisense strand sequences selected from the following, and the 5' end of the sense strand is conjugated to the ligand Tri-SM6.1 via a phosphate thioester or phosphodiester bond:

[0519] (1) Justice chain: SEQ ID NO:399, Antisense chain: SEQ ID NO:400;

[0520] (2) Justice chain: SEQ ID NO:479, Antisense chain: SEQ ID NO:480;

[0521] (3) Justice chain: SEQ ID NO:533, Antisense chain: SEQ ID NO:534;

[0522] (4) Justice chain: SEQ ID NO:597, Antisense chain: SEQ ID NO:598;

[0523] (5) Justice chain: SEQ ID NO:607, Antisense chain: SEQ ID NO:608;

[0524] (6) Justice chain: SEQ ID NO:617, Antisense chain: SEQ ID NO:618;

[0525] (7) Justice chain: SEQ ID NO:627, Antisense chain: SEQ ID NO:628;

[0526] The dsRNA (especially siRNA) of this invention can be conjugated to a target ligand via any linker known in the art. For example, the linker between siRNA and the target ligand can be a PEG linker, a saturated aliphatic alkyl chain, an unsaturated aliphatic alkyl chain, etc.

[0527] V.MUC5AC gene expression inhibition

[0528] The dsRNA of the present invention is capable of inhibiting MUC5AC gene expression by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%.

[0529] Inhibition of MUC5AC gene expression can be manifested by a reduction in the amount of mRNA expressed by a first cell or cell group (such cells may be present, for example, in a sample derived from a subject), wherein the MUC5AC gene is transcribed and the cell or these cells have been treated (e.g., by contacting the cell or these cells with the dsRNA of the present invention, or by administering the dsRNA of the present invention to a subject who currently or previously has these cells), such that MUC5AC gene expression is inhibited compared to a second cell or cell group (one or more control cells) that is substantially the same as the first cell or cell group but has not been so treated.

[0530] In some embodiments, the inhibition is assessed as a percentage of the mRNA level in the treated cells compared to the mRNA level in the control cells. Control cells or cell populations that can be used to assess inhibition of MUC5AC gene expression include cells or cell populations that have not been contacted with the dsRNA of the present invention. For example, the control cells or cell populations may be derived from individual subjects (e.g., human or animal subjects) prior to treatment with the dsRNA.

[0531] VI. Carriers and Cells

[0532] This disclosure provides vectors containing nucleotide sequences encoding the dsRNA described herein. The vectors of this invention are capable of amplifying and / or expressing the nucleotides encoding the dsRNA linked thereto. The vectors of this invention can be viruses, plasmids, or granules.

[0533] dsRNA targeting the MUC5AC gene can be expressed from transcriptional units inserted into DNA or RNA vectors. Expression can be transient (lasting hours to weeks) or persistent (lasting weeks to months or longer), depending on the specific construct used and the type of target tissue or cell. The coding nucleotides of the dsRNA targeting MUC5AC can be introduced into linear constructs, circular plasmids, or viral vectors. The nucleotides encoding the dsRNA targeting MUC5AC can be stably expressed by integration into the cellular genome or by stably inherited extrachromosomally. Generally, dsRNA expression vectors are typically DNA plasmids or viral vectors. Viral vector systems containing the coding sequence of the dsRNA targeting MUC5AC include, but are not limited to: (a) adenovirus vectors; (b) retrovirus vectors; (c) adeno-associated virus vectors; (d) herpes simplex virus vectors; (e) SV40 vectors; (f) polyomavirus vectors; (g) papillomavirus vectors; (h) microRNA virus vectors; or (i) poxvirus vectors.

[0534] This disclosure also provides cells containing the dsRNA or vector described herein. Any cell can be used as a host cell for the dsRNA or vector of this disclosure.

[0535] VII. Pharmaceutical Composition

[0536] This disclosure provides pharmaceutical compositions comprising the dsRNA, vector or cell described herein, and optionally a pharmaceutically acceptable carrier or excipient.

[0537] The dsRNA of the present invention can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically contain dsRNA and pharmaceutically acceptable carriers or excipients. Pharmaceutically acceptable carriers or excipients are those that facilitate the administration of dsRNA or its containing cells to the human body and / or promote its absorption or efficacy, such as: diluents, excipients like water, fillers like starch, sucrose, etc.; binders like cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone; humectants like glycerin; disintegrants like agar, calcium carbonate, and sodium bicarbonate; absorption enhancers like quaternary ammonium compounds; surfactants like hexadecyl alcohol; adsorbents like kaolin and soap clay; lubricants like talc, calcium / magnesium stearate, polyethylene glycol, etc. Other excipients may also be added to the composition.

[0538] The pharmaceutical compositions of the present invention may contain a pharmaceutically acceptable diluent or a sustained-release matrix in which the dsRNA of the present invention is embedded.

[0539] The pharmaceutical compositions of the present invention may comprise a drug delivery system for delivering dsRNA. The drug delivery systems of the present invention include, but are not limited to, nanoparticles (e.g., lipid nanoparticles, polymer-based nanoparticles), polymers, PEG, or cationic delivery systems, polylactic acid (PLA) microspheres, polylactic acid-glycolic acid copolymer (PLGA) microspheres, liposomes, micelles, inverse micelles, lipid cochleates, lipid microtubules, cholesterol, PEG lipids PEG-2000-C-DMG, PEG-2000-DMG (Moderna), ALC-0159, or DSPC.

[0540] In some embodiments, the dsRNA in the pharmaceutical compositions of the present invention may be contained in polymers and polymer-based nanoparticles.

[0541] In some specific embodiments, the polymer is based on poly(lactic-co-glycolic acid) copolymer (PLGA). In some specific embodiments, the PLGA-based polymer is modified to contain individual cationic groups.

[0542] In some specific embodiments, the polymer contains amine groups capable of becoming cationic, such as polyethyleneimine (PEI) and poly(L-lysine) (PLL), which can form a complex with dsRNA via electrostatic interactions and deliver dsRNA into cells. In some embodiments, PEG and PLL are chemically modified to improve in vivo potency and tolerability.

[0543] In some embodiments, the siRNA or cells in the pharmaceutical composition of the present invention can be delivered via the cationic polymer poly(β-amino ester) (PBAE).

[0544] In some embodiments, the pharmaceutical compositions of the present invention further comprise one or more additional (i.e., second, third, etc.) therapeutic agents. The additional therapeutic agent may be another MUC5AC RNAi agent (e.g., a MUC5AC RNAi agent targeting a different sequence within the MUC5AC gene). The additional therapeutic agent may also be a small molecule drug, an antibody, an antibody fragment, and / or an aptamer.

[0545] VIII. Reagent Kit

[0546] This disclosure provides kits containing the dsRNA, vector, or cells described herein.

[0547] The present invention also provides kits for using the dsRNA, vector, or cells described herein and / or performing the methods of the present invention. Such kits comprise one or more of the dsRNA or cells described herein, and may further include instructions for use. These instructions may describe methods for inhibiting MUC5AC gene expression in cells by contacting the cells with the dsRNA described herein in an amount that effectively inhibits MUC5AC gene expression.

[0548] In the case of in vitro contact between the dsRNA described in this invention and cells, the kit of this invention may optionally include a tool for contacting cells with the dsRNA described in this invention (e.g., an injection device) or a tool for measuring the inhibitory effect on the MUC5AC gene (e.g., a device for measuring the inhibition of MUC5AC mRNA or protein). Such a device for measuring the inhibition of the MUC5AC gene may include a device for obtaining a sample from a subject.

[0549] When administering the dsRNA of the present invention or cells that have already been introduced with dsRNA in vitro into the body, the kit of the present invention may optionally include means for administering the dsRNA or cells of the present invention to a subject or means for determining a therapeutically effective amount or a preventatively effective amount.

[0550] IX. Methods and Uses

[0551] This invention provides a method for inhibiting MUC5AC gene expression in cells, the method comprising contacting the cells with a dsRNA or pharmaceutical composition as described herein. In some embodiments, the method is performed in vitro. In some embodiments, the method is performed in vivo. This invention provides the dsRNA or pharmaceutical composition described herein for inhibiting MUC5AC gene expression in cells.

[0552] This invention provides a method for inhibiting MUC5AC gene expression in the cells of a subject, the method comprising administering to the subject the dsRNA, vector, cell, or pharmaceutical composition described herein. This invention provides the dsRNA, vector, cell, or pharmaceutical composition described herein for inhibiting MUC5AC gene expression in the cells of a subject. This invention provides the use of the dsRNA, vector, cell, or pharmaceutical composition described herein in the preparation of a medicament for inhibiting MUC5AC gene expression in the cells of a subject.

[0553] This invention also provides a method for treating a disease or condition in which a subject benefits from reduced MUC5AC gene expression, the method comprising administering to the subject an effective amount of the dsRNA, vector, cell, or pharmaceutical composition described herein. This invention also provides the dsRNA, vector, cell, or pharmaceutical composition described herein for treating a disease or condition in which a subject benefits from reduced MUC5AC gene expression. This invention further provides the use of the dsRNA, vector, cell, or pharmaceutical composition described herein in the preparation of a medicament for treating a disease or condition in which a subject benefits from reduced MUC5AC gene expression.

[0554] In some embodiments, the disease or condition that benefits from reduced MUC5AC gene expression is a disease mediated or associated with MUC5AC expression. In some embodiments, the disease mediated or associated with MUC5AC expression is a respiratory disease. In some embodiments, the disease mediated or associated with MUC5AC expression is a mucus obstructive airway disease, preferably selected from asthma, COPD, and bronchiectasis, more preferably selected from asthma and bronchiectasis.

[0555] In some embodiments, the dsRNA, vector, cell, or pharmaceutical composition described herein can be administered in combination with one or more additional (i.e., a second, third, etc.) therapeutic agents. The additional therapeutic agent may be another MUC5AC RNAi agent (e.g., a MUC5AC RNAi agent targeting a different sequence within the MUC5AC gene). The additional therapeutic agent may also be a small molecule drug, antibody, antibody fragment, and / or aptamer.

[0556] In some implementations, the dose administered to the subject may vary depending on the implementation method, the drug used, the method of administration, and the site of treatment and the subject. However, the dose should be sufficient to provide a therapeutic response. Clinicians can determine the effective amount to administer to humans or other subjects to treat a medical condition. The precise amount required for effective treatment may depend on many factors, such as the activity of the dsRNA and the route of administration.

[0557] The dosage of the dsRNA, vector, cell, or pharmaceutical composition described herein can be administered to mammals in a single dose or in a series of sub-dose over an appropriate period of time, such as daily, bi-weekly, weekly, bi-weekly, bi-weekly, bi-monthly, semi-annually, or annually as needed. Dosage units containing an effective amount of the dsRNA, vector, cell, or pharmaceutical composition can be administered as a single daily dose, or the total daily dose can be administered as needed in two, three, four, or more daily fractions.

[0558] The appropriate method of administration can be chosen by the physician. In some embodiments, the dsRNA, vector, cell, or pharmaceutical composition described herein is administered by inhalation, intranasal administration, intratracheal administration, or oropharyngeal aspiration. In some embodiments, the pharmaceutical composition can be administered by injection, such as intravenous, intramuscular, intradermal, subcutaneous, intra-articular, intraocular, intraperitoneal, or local administration. For example, in some embodiments, it is desirable to inhibit the expression of the MUC5AC gene in the lung epithelium using the dsRNA, vector, cell, or pharmaceutical composition described herein, and for this purpose, administration by inhalation (e.g., via an inhaler device, such as a metered-dose inhaler, or a nebulizer such as a jet or vibrating mesh nebulizer, or a soft mist inhaler) is particularly suitable and advantageous. In a preferred embodiment, the dsRNA described herein is administered via airway nebulization. The dosage and method of administration can vary depending on the subject's weight, age, condition, etc., and can be appropriately selected.

[0559] In some embodiments, the subject is a mammal. In some embodiments, the subject is a primate mammal. In some embodiments, the subject is a human.

[0560] X. Example

[0561] The following embodiments are provided for the purpose of illustrating various embodiments of the invention, but are not intended to limit the invention in any way. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Variations and other uses will be apparent to those skilled in the art within the spirit of the invention as defined by the claims. Experimental methods not specifically described in the following embodiments were performed under conventional conditions in the art or as recommended by the manufacturer. Unless otherwise stated, the experimental materials and reagents used in the following embodiments are commercially available.

[0562] Example 1: siRNA design and synthesis

[0563] Based on the human MUC5AC gene transcript (GenBank NM_001304359.2), siRNA sequences were designed at different sites. Specific sequence information is shown in Table 1. Among them, siPC-1, the dominant sequence targeting MUC5AC gene position 4993 disclosed in WO2022251394A1, was used as a positive control siRNA.

[0564] The siRNA duplexes of this application are prepared using techniques known in the art, such as conventional solid-phase nucleic acid synthesis. The polynucleotides of the siRNA can be assembled using standard nucleotides or nucleoside precursors (e.g., phosphoramide) on a suitable nucleic acid synthesizer via deprotection, coupling, oxidation, and capping processes. Custom synthesis of siRNA can also be obtained from commercial suppliers.

[0565] Table 1. siRNA sequences targeting MUC5AC

[0566] Example 2: Inhibition of human MUC5AC expression in A549 cells by siRNA sequence (screening for inhibition activity at two concentration points)

[0567] The effects of the candidate siRNA duplex sequences shown in Table 1 on MUC5AC mRNA expression levels were tested in vitro. A549 cells were cultured in F-12K medium containing 10% fetal bovine serum at 37°C with 5% CO2. Cells were seeded and transfected when they were in the logarithmic growth phase and in good condition. Cell density was adjusted to approximately 7000 cells per well in 96-well plates.

[0568] Following the product manual, siRNA was transfected using Lipofectamine RNAiMAX (ThermoFisher) at final concentrations of 10 nM and 1 nM. After incubation at 37°C with 5% CO2 for 48 h, cells were collected, and total RNA was extracted using a tissue / cell total RNA extraction kit (YEASEN). Reverse transcription was then performed using the HiScript III 1st Strand cDNA Synthesis Kit (YEASEN), or cDNA was prepared directly from cells using a Cells-to-cDNA kit (Vazyme). Finally, qPCR was performed using the QuantStudio Real-time PCR system (384 well). After PCR, relative quantification was performed using the 2-ΔΔCt method with an internal reference gene as the standard. The results were normalized to the Lipo-transfected A549 cells to obtain the residual mRNA expression level and calculated relative to the positive control siPC-1. The results are shown in Tables 2 and 3. Table 2 lists the detected siRNA duplex numbers and average residual mRNA expression levels. Table 3 lists the double-stranded siRNAs detected and their relative expression levels relative to siPC-1. The lower the relative ratio, the lower the residual expression level of MUC5AC mRNA, i.e., the higher the inhibitory activity of the siRNA.

[0569] As shown in Table 3, at the 10 nM concentration level, the residual mRNA expression level of 47 siRNA duplexes was lower than that of the positive control siPC-1 (i.e., the relative ratio was less than 1); at the 1 nM concentration level, the residual mRNA expression level of 29 siRNA duplexes was lower than that of the positive control siPC-1; and at both the 10 nM and 1 nM concentration levels, the residual mRNA expression level of 28 siRNA duplexes was lower than that of the positive control siPC-1.

[0570] Table 2. Inhibition of MUC5AC mRNA expression levels by siRNA sequences

[0571] Table 3. Residual expression levels of MUC5AC mRNA relative to the siPC-1 treatment group

[0572] Example 3: Inhibition of human MUC5AC expression in A549 cells by siRNA sequence (screening for inhibition activity at two concentration points)

[0573] Candidate sequences shown in Table 1 were screened in A549 cells using two concentrations (10 nM and 0.1 nM). A549 cells were cultured in F-12K medium containing 10% fetal bovine serum at 37°C with 5% CO2. Once the cells were in the logarithmic growth phase and in good condition, they were seeded and transfected. Cell density was adjusted to approximately 2000 cells per well in a 96-well plate.

[0574] Following the product instruction manual, siRNA was transfected using Lipofectamine RNAiMAX (ThermoFisher) at final siRNA concentrations of 10 nM and 0.1 nM. After 48 hours of treatment, cells were collected, and other procedures were performed as in Example 2.

[0575] After the PCR reaction, relative quantification was performed using the 2-ΔΔCt method with the internal reference gene as the standard. The results were normalized to the Lipo-transfected A549 cells to obtain the residual mRNA expression level, and the relative expression level relative to the positive control siPC-1 was calculated. The results are shown in Tables 4 and 5. Table 4 lists the detected siRNA duplex numbers and average residual mRNA expression levels. Table 5 lists the detected siRNA duplex numbers and their relative expression levels relative to siPC-1. A lower relative ratio indicates a lower residual MUC5AC mRNA expression level, meaning higher siRNA inhibitory activity.

[0576] As shown in Table 5, at a concentration of 10 nM, the residual expression level of MUC5AC mRNA in 49 siRNA duplexes in A549 cells was lower than that in the positive control siPC-1 (i.e., the relative ratio was less than 1); at a concentration of 0.1 nM, the residual expression level of MUC5AC mRNA in A549 cells was lower than that in the positive control siPC-1; and the residual expression level of MUC5AC mRNA in 46 siRNA duplexes was lower than that in the positive control siPC-1 at both concentration levels.

[0577] Table 4. Inhibition of MUC5AC mRNA expression levels by siRNA sequences

[0578] Table 5. Residual expression levels of MUC5AC mRNA relative to the siPC-1 treatment group

[0579] Example 4: IC50 value of A549 transfected cells

[0580] The preferred sequences from Examples 2 and 3, where the residual expression level of MUC5AC mRNA was lower than that of the positive control siPC-1 and showed high homology with human and monkey cells, were serially diluted with Nuclease-Free Water to different concentrations and transfected into A549 cells. The transfection and quantitative PCR detection and analysis procedures were the same as in Example 2. The IC50 value of siRNA inhibiting MUC5AC mRNA expression was calculated using Graphad Prism software, and the results are shown in Table 6.

[0581] As shown in Table 6, the IC50 values ​​of the tested preferred sequences were all lower than those of the positive control siPC-1 treatment group. For example, the IC50 value of TD-U-98 in A549 cells was only 0.045 times that of the positive control siPC-1.

[0582] Table 6. IC50 values ​​of siRNA inhibiting MUC5AC mRNA expression in A549 cells.

[0583] Example 5: Sequence Modification

[0584] TD-U-86, TD-U-87, TD-U-88, TD-U-91, TD-U-92, TD-U-93, TD-U-95, TD-U-96, TD-U-98, TD-U-100, TD-U-102, TD-U-106, TD-U-107, TD-U-109, TD-U-117, TD-U-119, TD-U-123, TD-U-133, TD-U-147 and TD-U-162 were chemically modified using the modification methods M01 to M10 shown in Table 7. The modified sequences are shown in Table 8. TD-U-174, TD-U-187, TD-U-235, TD-U-245, and TD-U-265 were chemically modified using the modification methods M01 to M13 shown in Table 7. The modified sequences are shown in Table 8.

[0585] The control siRNA in Example 1 The modified sequence obtained by modifying siPC-1 serves as a positive control (referred to as siPC-1M). The specific sequence information for siPC-1M is as follows:

[0586] Chain of Justice (5'-3'): (invAb)sGmCmUmGmAmUmUfUmGfCmCfUmGmAmAmCmAmAmGmAmAms(invAb)(SEQ IN NO:809)

[0587] Antisense chain (5'-3'): cPrpuUfCmUmUmGmUmUmCmAmGmGfCmAfAmAfUmCmAmGmsCm (SEQ INNO:810)

[0588] Where Nm represents 2'-O-methyl modified adenine (A), cytosine (C), uracil (U), and guanine (G), Nf represents 2'-fluoro modified adenine (A), cytosine (C), uracil (U), and guanine (G), VP represents 5'-(E)-vinylphosphonate, s represents thiophosphate linkage, dN represents deoxyribonucleotide modification (U is modified to dT; A is modified to dA; C is modified to dC; G is modified to dG); invAb represents reverse abase-free deoxyribose residues; and cPrpu represents 5'-cyclopropylphosphonate-2'-O-methyluridine.

[0589] The chemically modified siRNA of this application is prepared using techniques known in the art, such as conventional nucleic acid solid-phase synthesis techniques. The polynucleotides of the siRNA can be assembled on a suitable nucleic acid synthesizer using standard nucleotides or nucleoside precursors (e.g., phosphoramide). Custom synthesis of siRNA can also be obtained from commercial suppliers.

[0590] Table 7. Modification Methods

[0591] Table 8. Modified Sequence Information Table

[0592] Example 6: Inhibition of human MUC5AC expression in A549 cells by modified siRNA sequences (screening at 10 nM and 1 nM concentrations)

[0593] The effects of the partially modified siRNA double-stranded sequences shown in Table 8 on MUC5AC mRNA expression levels at concentrations of 10 nM and 1 nM were tested in vitro. The specific experimental procedures were the same as in Example 2. Modified siPC-1M was used as a positive control.

[0594] Table 9 lists the double-stranded siRNAs detected and their average residual mRNA expression levels. Lower residual MUC5AC mRNA expression levels indicate higher siRNA inhibitory activity.

[0595] As shown in Table 9, all tested sequences inhibited the expression of MUC5AC mRNA at both 10 nM and 1 nM concentrations. Specifically, at 10 nM, 186 sequences were superior to siPC-1M; and at 1 nM, 120 sequences were superior to siPC-1M.

[0596] Table 9. Inhibition of MUC5AC mRNA expression levels by modified siRNA sequences

[0597] Example 7: Inhibition of human MUC5AC expression in A549 cells by modified siRNA sequences (screening at 10 nM and 0.1 nM concentrations)

[0598] The effects of the partially modified siRNA double-stranded sequences shown in Table 8 on the expression levels of MUC5AC mRNA at concentrations of 10 nM and 0.1 nM were tested in vitro. The specific experimental procedures were the same as in Example 3. Modified siPC-1M was used as a positive control.

[0599] Table 10 lists the duplex numbers of the detected siRNAs and their average residual mRNA expression levels. The lower the residual MUC5AC mRNA expression level, the higher the inhibitory activity of the siRNA.

[0600] As shown in Table 10, all tested sequences inhibited the expression of MUC5AC mRNA at both 10 nM and 0.1 nM concentrations. Among them, 40 sequences were superior to siPC-1M at 10 nM concentration, and 10 sequences were superior to siPC-1M at 0.1 nM concentration.

[0601] Table 10. Inhibition of MUC5AC mRNA expression levels by modified siRNA sequences

[0602] Example 8: Determination of IC50 value of modified siRNA sequence

[0603] The IC50 values ​​of some of the dominant sequences from Examples 6 and 7 were determined in A549 cells. The lyophilized sequence powders were serially diluted with Nuclease-Free Water to different concentrations and transfected into A549 cells. The transfection and quantitative PCR detection analysis procedures were the same as in Example 2. The IC50 values ​​of siRNA inhibiting MUC5AC mRNA expression were calculated using Graphad Prism software, and the results are shown in Table 11.

[0604] Table 11 shows that 17 sequences exhibited superior activity compared to the positive control siPC-1M. In particular, the IC50 values ​​of sequences TD-95-M01, TD-102-M01, TD-107-M08, TD-119-M10, TD-123-M05, and TD-123-M10 were 90.65%, 91.96%, 82.95%, 81.80%, 96.06%, and 96.88% higher than the positive control, respectively.

[0605] Table 11. IC50 values ​​of modified sequences

[0606] Example 9: Preparation of siRNA with targeted ligand conjugation

[0607] TD-95-M01, TD-102-M01, TD-107-M08, TD-119-M10, TD-123-M05, TD-123-M10, and TD-133-M05 were respectively ligated to the αvβ6 epithelial cell targeting ligand “Tri-SM6.1” via a tridentate linker at the 5' end of the positive-sense strand. After ligation, they were named Tri-SM6.1-95M01, Tri-SM6.1-102M01, Tri-SM6.1-107M08, Tri-SM6.1-119M10, Tri-SM6.1-123M05, Tri-SM6.1-123M10, and Tri-SM6.1-133M05, respectively. As a control, the ligand “Tri-SM6.1” was liganded to the 5' end of the positive chain of siPC-M1, and the resulting compound was named Tri-SM6.1-siPC-1M. The structure of the ligand molecule “Tri-SM6.1” is shown below:

[0608] The ligand molecule “Tri-SM6.1” can be prepared according to the method described in WO2022251394A1.

[0609] Example 10: Activity of ligand-targeted siRNA in cynomolgus monkeys

[0610] In the in vivo experiment, three male cynomolgus monkeys were used in each group. On day 1, 2 mg / kg of candidate siRNA molecules Tri-SM6.1-95M01, Tri-SM6.1-102M01, Tri-SM6.1-107M08, Tri-SM6.1-119M10, Tri-SM6.1-123M05, Tri-SM6.1-123M10, and Tri-SM6.1-133M05 were delivered to the lungs via nebulized injection. The nebulization volume was 0.5 mL / kg. Twenty-one days after the single administration, the monkeys were anesthetized and sacrificed. Tracheal tubes at all levels were collected, homogenized, and the expression level of MUC5AC mRNA was determined by RT-PCR. Tri-SM6.1-siPC-1M from Example 9 was used as a positive control, and the saline group was used as a negative control. Relative quantitative analysis was performed using the internal reference gene as a standard, and the saline blank control group was normalized. The results are shown in Table 12 below. Table 12 below shows the residual expression levels of MUC5AC mRNA in the trachea, main bronchus, lobar bronchus, and segmental bronchus.

[0611] Table 12. Residual expression level of MUC5AC mRNA in cynomolgus monkeys 21 days after a single nebulized drug administration

[0612] mRNA expression results showed that all drug-treated groups effectively reduced the expression level of MUC5AC mRNA in cynomolgus monkeys. Furthermore, the Tri-SM6.1-95M01, Tri-SM6.1-102M01, Tri-SM6.1-107M08, Tri-SM6.1-119M10, Tri-SM6.1-123M05, Tri-SM6.1-123M10, and Tri-SM6.1-133M05 groups showed a greater reduction in mRNA levels than the positive control Tri-SM6.1-siPC-1M group.

Claims

1. A double-stranded RNA (dsRNA) for inhibiting MUC5AC gene expression, said dsRNA comprising a sense strand and an antisense strand forming a double-stranded region, wherein the length of the sense strand and the antisense strand are each independently 15-30 nucleotides, and said antisense strand comprises SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 9 2, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 16 0, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.

2. The dsRNA of claim 1, wherein the sense strand comprises SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 9 1, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

3. The dsRNA of claim 1, wherein the antisense strand comprises SEQ ID NO: 4, 6, 8, 10, 12, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 56, 58, 60, 62, 64, 66, 68, 70, 74, 76, 78, 80, 82, 86, 92, 96, 98, 100, 102, 106, 110, 112, 116, 128, 130, 134, 152, 156, 158, 160, 162, 164, 166, 168, 170, 174, 176, 178, 180, 182, 184, 186 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.

4. The dsRNA of claim 3, wherein the sense strand comprises SEQ ID NO: 3, 5, 7, 9, 11, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 55, 57, 59, 61, 63, 65, 67, 69, 73, 75, 77, 79, 81, 85, 91, 95, 97, 99, 101, 105, 109, 111, 115, 127, 129, 133, 151, 155, 157, 159, 161, 163, 165, 167, 169, 173, 175, 177, 179, 181, 183, 185, A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

5. The dsRNA of claim 1, wherein the antisense strand comprises SEQ ID NO: 4, 6, 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 62, 64, 66, 68, 74, 80, 82, 86, 102, 128, 156, 158, 162, 166, 168, 170, 176, 178, 182, 184, 186, 188, 190, 192, 194 A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 196, 198, 204, 206, 208, 212, 214, 218, 220, 228, 230, 234, 236, 238, 240, 242, 244, 250, 252, 254, 256, 260, 264, 266, 268, 270, 272, 274, 278.

6. The dsRNA of claim 5, wherein the sense strand comprises SEQ ID NO: 3, 5, 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 61, 63, 65, 67, 73, 79, 81, 85, 101, 127, 155, 157, 161, 165, 167, 169, 175, 177, 181, 183, 185, 187, 189, 191, 193, A nucleotide sequence of at least 15 consecutive nucleotides of any one of the nucleotide sequences shown in 195, 197, 203, 205, 207, 211, 213, 217, 219, 227, 229, 233, 235, 237, 239, 241, 243, 249, 251, 253, 255, 259, 263, 265, 267, 269, 271, 273, 277.

7. The dsRNA of claim 1, wherein the antisense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 8, 10, 12, 16, 18, 20, 24, 26, 30, 32, 34, 38, 40, 44, 58, 60, 66, 86, 102, 128, 144, 156, 184, 192, 278.

8. The dsRNA of claim 7, wherein the sense strand comprises a nucleotide sequence of at least 15 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 7, 9, 11, 15, 17, 19, 23, 25, 29, 31, 33, 37, 39, 43, 57, 59, 65, 85, 101, 127, 143, 155, 183, 191, 277.

9. The dsRNA of any one of claims 1-8, wherein the dsRNA is siRNA.

10. The dsRNA of any one of claims 1-9, wherein the length of the sense strand and the antisense strand is each independently 15-27 nucleotides, preferably 18-25 nucleotides, more preferably 19-23 nucleotides.

11. The dsRNA according to any one of claims 1-10, wherein the antisense strand comprises SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 9 6, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 16 6, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 2 A nucleotide sequence of at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleotides of any one of the nucleotide sequences shown in 36, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.Preferably, the antisense chain comprises SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88. 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 15 6, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220 The nucleotide sequence shown in any one of the following: 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278.

12. The dsRNA according to any one of claims 1-11, wherein the sense strand comprises SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 9 3, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 1 63, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 2 A nucleotide sequence of at least 16, at least 17, or at least 18 consecutive nucleotides of any one of the nucleotide sequences shown in 31, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, and 277.Preferably, the justice chain comprises SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87. 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219 The nucleotide sequence shown is any one of the following: 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277.

13. The dsRNA of any one of claims 1-12, wherein the dsRNA comprises any one of the following pairs of paired sense and antisense sequences: (1) Justice chain: SEQ ID NO:3, Antisense chain: SEQ ID NO:4; (2) Justice chain: SEQ ID NO:5, Antisense chain: SEQ ID NO:6; (3) Justice chain: SEQ ID NO:7, Antisense chain: SEQ ID NO:8; (4) Justice chain: SEQ ID NO:9, Antisense chain: SEQ ID NO:10; (5) Justice chain: SEQ ID NO:11, Antisense chain: SEQ ID NO:12; (6) Justice chain: SEQ ID NO:13, Antisense chain: SEQ ID NO:14; (7) Justice chain: SEQ ID NO:15, Antisense chain: SEQ ID NO:16; (8) Justice Chain: SEQ ID NO:17, Antisense Chain: SEQ ID NO:18; (9) Justice Chain: SEQ ID NO:19, Antisense Chain: SEQ ID NO:20; (10) Justice chain: SEQ ID NO:21, Antisense chain: SEQ ID NO:22; (11) Justice chain: SEQ ID NO:23, Antisense chain: SEQ ID NO:24; (12) Justice chain: SEQ ID NO:25, Antisense chain: SEQ ID NO:26; (13) Justice chain: SEQ ID NO:27, Antisense chain: SEQ ID NO:28; (14) Justice chain: SEQ ID NO:29, Antisense chain: SEQ ID NO:30; (15) Justice chain: SEQ ID NO:31, Antisense chain: SEQ ID NO:32; (16) Justice chain: SEQ ID NO:33, Antisense chain: SEQ ID NO:34; (17) Justice chain: SEQ ID NO:35, Antisense chain: SEQ ID NO:36; (18) Justice chain: SEQ ID NO:37, Antisense chain: SEQ ID NO:38; (19) Justice Chain: SEQ ID NO:39, Antisense Chain: SEQ ID NO:40; (20) Justice chain: SEQ ID NO:41, Antisense chain: SEQ ID NO:42; (21) Justice chain: SEQ ID NO:43, Antisense chain: SEQ ID NO:44; (22) Justice chain: SEQ ID NO:45, Antisense chain: SEQ ID NO:46; (23) Justice chain: SEQ ID NO:47, Antisense chain: SEQ ID NO:48; (24) Justice chain: SEQ ID NO:49, Antisense chain: SEQ ID NO:50; (25) Justice chain: SEQ ID NO:51, Antisense chain: SEQ ID NO:52; (26) Justice chain: SEQ ID NO:53, Antisense chain: SEQ ID NO:54; (27) Justice chain: SEQ ID NO:55, Antisense chain: SEQ ID NO:56; (28) Justice chain: SEQ ID NO:57, Antisense chain: SEQ ID NO:58; (29) Justice chain: SEQ ID NO:59, Antisense chain: SEQ ID NO:60; (30) Justice chain: SEQ ID NO:61, Antisense chain: SEQ ID NO:62; (31) Justice chain: SEQ ID NO:63, Antisense chain: SEQ ID NO:64; (32) Justice chain: SEQ ID NO:65, Antisense chain: SEQ ID NO:66; (33) Justice chain: SEQ ID NO:67, Antisense chain: SEQ ID NO:68; (34) Justice chain: SEQ ID NO:69, Antisense chain: SEQ ID NO:70; (35) Justice chain: SEQ ID NO:71, Antisense chain: SEQ ID NO:72; (36) Justice chain: SEQ ID NO:73, Antisense chain: SEQ ID NO:74; (37) Justice chain: SEQ ID NO:75, Antisense chain: SEQ ID NO:76; (38) Justice chain: SEQ ID NO:77, Antisense chain: SEQ ID NO:78; (39) Justice chain: SEQ ID NO:79, Antisense chain: SEQ ID NO:80; (40) Justice chain: SEQ ID NO:81, Antisense chain: SEQ ID NO:82; (41) Justice chain: SEQ ID NO:83, Antisense chain: SEQ ID NO:84; (42) Justice chain: SEQ ID NO:85, Antisense chain: SEQ ID NO:86; (43) Justice chain: SEQ ID NO:87, Antisense chain: SEQ ID NO:88; (44) Justice chain: SEQ ID NO:89, Antisense chain: SEQ ID NO:90; (45) Justice chain: SEQ ID NO:91, Antisense chain: SEQ ID NO:92; (46) Justice chain: SEQ ID NO:93, Antisense chain: SEQ ID NO:94; (47) Justice chain: SEQ ID NO:95, Antisense chain: SEQ ID NO:96; (48) Justice chain: SEQ ID NO:97, Antisense chain: SEQ ID NO:98; (49) Justice Chain: SEQ ID NO:99, Antisense Chain: SEQ ID NO:100; (50) Justice chain: SEQ ID NO:101, Antisense chain: SEQ ID NO:102; (51) Justice chain: SEQ ID NO:103, Antisense chain: SEQ ID NO:104; (52) Justice chain: SEQ ID NO:105, Antisense chain: SEQ ID NO:106; (53) Justice chain: SEQ ID NO:107, Antisense chain: SEQ ID NO:108; (54) Justice chain: SEQ ID NO:109, Antisense chain: SEQ ID NO:110; (55) Justice chain: SEQ ID NO:111, Antisense chain: SEQ ID NO:112; (56) Justice chain: SEQ ID NO:113, Antisense chain: SEQ ID NO:114; (57) Justice chain: SEQ ID NO:115, Antisense chain: SEQ ID NO:116; (58) Justice chain: SEQ ID NO:117, Antisense chain: SEQ ID NO:118; (59) Justice chain: SEQ ID NO:119, Antisense chain: SEQ ID NO:120; (60) Justice chain: SEQ ID NO:121, Antisense chain: SEQ ID NO:122; (61) Justice chain: SEQ ID NO:123, Antisense chain: SEQ ID NO:124; (62) Justice chain: SEQ ID NO:125, Antisense chain: SEQ ID NO:126; (63) Justice chain: SEQ ID NO:127, Antisense chain: SEQ ID NO:128; (64) Justice chain: SEQ ID NO:129, Antisense chain: SEQ ID NO:130; (65) Justice chain: SEQ ID NO:131, Antisense chain: SEQ ID NO:132; (66) Justice chain: SEQ ID NO:133, Antisense chain: SEQ ID NO:134; (67) Justice chain: SEQ ID NO:135, Antisense chain: SEQ ID NO:136; (68) Justice chain: SEQ ID NO:137, Antisense chain: SEQ ID NO:138; (69) Justice chain: SEQ ID NO:139, Antisense chain: SEQ ID NO:140; (70) Justice chain: SEQ ID NO:141, Antisense chain: SEQ ID NO:142; (71) Justice chain: SEQ ID NO:143, Antisense chain: SEQ ID NO:144; (72) Justice chain: SEQ ID NO:145, Antisense chain: SEQ ID NO:146; (73) Justice chain: SEQ ID NO:147, Antisense chain: SEQ ID NO:148; (74) Justice chain: SEQ ID NO:149, Antisense chain: SEQ ID NO:150; (75) Justice chain: SEQ ID NO:151, Antisense chain: SEQ ID NO:152; (76) Justice chain: SEQ ID NO:153, Antisense chain: SEQ ID NO:154; (77) Justice chain: SEQ ID NO:155, Antisense chain: SEQ ID NO:156; (78) Justice chain: SEQ ID NO:157, Antisense chain: SEQ ID NO:158; (79) Justice chain: SEQ ID NO:159, Antisense chain: SEQ ID NO:160; (80) Justice chain: SEQ ID NO:161, Antisense chain: SEQ ID NO:162; (81) Justice chain: SEQ ID NO:163, Antisense chain: SEQ ID NO:164; (82) Justice chain: SEQ ID NO:165, Antisense chain: SEQ ID NO:166; (83) Justice chain: SEQ ID NO:167, Antisense chain: SEQ ID NO:168; (84) Justice chain: SEQ ID NO:169, Antisense chain: SEQ ID NO:170; (85) Justice chain: SEQ ID NO:171, Antisense chain: SEQ ID NO:172; (86) Justice chain: SEQ ID NO:173, Antisense chain: SEQ ID NO:174; (87) Justice chain: SEQ ID NO:175, Antisense chain: SEQ ID NO:176; (88) Justice Chain: SEQ ID NO:177, Antisense Chain: SEQ ID NO:178; (89) Justice chain: SEQ ID NO:179, Antisense chain: SEQ ID NO:180; (90) Justice chain: SEQ ID NO:181, Antisense chain: SEQ ID NO:182; (91) Justice chain: SEQ ID NO:183, Antisense chain: SEQ ID NO:184; (92) Justice chain: SEQ ID NO:185, Antisense chain: SEQ ID NO:186; (93) Justice chain: SEQ ID NO:187, Antisense chain: SEQ ID NO:188; (94) Justice Chain: SEQ ID NO:189, Antisense Chain: SEQ ID NO:190; (95) Justice chain: SEQ ID NO:191, Antisense chain: SEQ ID NO:192; (96) Justice Chain: SEQ ID NO:193, Antisense Chain: SEQ ID NO:194; (97) Justice chain: SEQ ID NO:195, Antisense chain: SEQ ID NO:196; (98) Justice Chain: SEQ ID NO:197, Antisense Chain: SEQ ID NO:198; (99) Justice Chain: SEQ ID NO:199, Antisense Chain: SEQ ID NO:200; (100) Justice chain: SEQ ID NO:201, Antisense chain: SEQ ID NO:202; (101) Justice chain: SEQ ID NO:203, Antisense chain: SEQ ID NO:204; (102) Justice chain: SEQ ID NO:205, Antisense chain: SEQ ID NO:206; (103) Justice chain: SEQ ID NO:207, Antisense chain: SEQ ID NO:208; (104) Justice Chain: SEQ ID NO:209, Antisense Chain: SEQ ID NO:210; (105) Justice chain: SEQ ID NO:211, Antisense chain: SEQ ID NO:212; (106) Justice chain: SEQ ID NO:213, Antisense chain: SEQ ID NO:214; (107) Justice chain: SEQ ID NO:215, Antisense chain: SEQ ID NO:216; (108) Justice Chain: SEQ ID NO:217, Antisense Chain: SEQ ID NO:218; (109) Justice Chain: SEQ ID NO:219, Antisense Chain: SEQ ID NO:220; (110) Justice chain: SEQ ID NO:221, Antisense chain: SEQ ID NO:222; (111) Justice chain: SEQ ID NO:223, Antisense chain: SEQ ID NO:224; (112) Justice chain: SEQ ID NO:225, Antisense chain: SEQ ID NO:226; (113) Justice chain: SEQ ID NO:227, Antisense chain: SEQ ID NO:228; (114) Justice chain: SEQ ID NO:229, Antisense chain: SEQ ID NO:230; (115) Justice chain: SEQ ID NO:231, Antisense chain: SEQ ID NO:232; (116) Justice chain: SEQ ID NO:233, Antisense chain: SEQ ID NO:234; (117) Justice chain: SEQ ID NO:235, Antisense chain: SEQ ID NO:236; (118) Justice chain: SEQ ID NO:237, Antisense chain: SEQ ID NO:238; (119) Justice chain: SEQ ID NO:239, Antisense chain: SEQ ID NO:240; (120) Justice chain: SEQ ID NO:241, Antisense chain: SEQ ID NO:242; (121) Justice chain: SEQ ID NO:243, Antisense chain: SEQ ID NO:244; (122) Justice chain: SEQ ID NO:245, Antisense chain: SEQ ID NO:246; (123) Justice chain: SEQ ID NO:247, Antisense chain: SEQ ID NO:248; (124) Justice chain: SEQ ID NO:249, Antisense chain: SEQ ID NO:250; (125) Justice chain: SEQ ID NO:251, Antisense chain: SEQ ID NO:252; (126) Justice chain: SEQ ID NO:253, Antisense chain: SEQ ID NO:254; (127) Justice chain: SEQ ID NO:255, Antisense chain: SEQ ID NO:256; (128) Justice chain: SEQ ID NO:257, Antisense chain: SEQ ID NO:258; (129) Justice chain: SEQ ID NO:259, Antisense chain: SEQ ID NO:260; (130) Justice chain: SEQ ID NO:261, Antisense chain: SEQ ID NO:262; (131) Justice chain: SEQ ID NO:263, Antisense chain: SEQ ID NO:264; (132) Justice chain: SEQ ID NO:265, Antisense chain: SEQ ID NO:266; (133) Justice chain: SEQ ID NO:267, Antisense chain: SEQ ID NO:268; (134) Justice chain: SEQ ID NO:269, Antisense chain: SEQ ID NO:270; (135) Justice chain: SEQ ID NO:271, Antisense chain: SEQ ID NO:272; (136) Justice chain: SEQ ID NO:273, Antisense chain: SEQ ID NO:274; (137) Justice chain: SEQ ID NO:275, Antisense chain: SEQ ID NO:276; (138) Justice chain: SEQ ID NO:277, Antisense chain: SEQ ID NO:

278.

14. The dsRNA of any one of claims 1-13, wherein at least one nucleotide of the sense strand and / or the antisense strand is a modified nucleotide.

15. The dsRNA of any one of claims 1-14, wherein substantially all nucleotides of the sense strand and substantially all nucleotides of the antisense strand are modified nucleotides, or wherein all nucleotides of the sense strand and all nucleotides of the antisense strand are modified nucleotides.

16. The dsRNA of any one of claims 1-15, wherein the sense strand and the antisense strand each independently comprise one or more nucleotide modifications selected from the group consisting of: 2'-O-methyl modification, 2'-O-methoxyethyl modification, 2'-fluoro modification, DNA modification, glycerol nucleic acid (GNA) modification, unlocking nucleic acid (UNA) modification, vinylphosphonate modification (e.g., 5'-VP modification), inverse non-base deoxyribonucleotide (invAb) modification, and phosphate thioester nucleotide interbonding modification; preferably, wherein the sense strand and the antisense strand each independently comprise one or more nucleotide modifications selected from the group consisting of: 2'-O-methyl modification, 2'-fluoro modification, DNA modification, phosphate thioester nucleotide interbonding modification, 5'-VP modification, and invAb modification.

17. The dsRNA of any one of claims 1-16, wherein the antisense strand has a length of 23 nucleotides and the sense strand has a length of 21 nucleotides.

18. The dsRNA of claim 17, wherein the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 5, 11, 13, 15, 17, 19, 21, 22, 23, counted from the 5' end; preferably, wherein the antisense strand has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 13, 15, 17, 19, 20, 21, 22, 23, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 2, 14, and 16.

19. The dsRNA of claim 17 or 18, wherein the antisense strand has: (i) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 12, 14, and 16; or (ii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 7, 8, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 9, 12, 14, and 16; or (iii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 5, 6, 7, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 4, 12, 14, and 16; or (iv) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 10, 14, and 16; or (v) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20; or (vi) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 19, 20, 21, 22, and 23, and the nucleotides with 2'-fluoro modification at positions 2, 6, 14, 16, and 18; or (vii) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the nucleotides with 2'-fluoro modification at positions 2, 8, 9, 14, and 16; or (viii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 7, 14, and 16; or (ix) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 14, and 16; or (x) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, and the nucleotides with 2'-fluorine modification at positions 2, 7, 12, 14, and 16; and / or (xi) Starting from the 5' end, count the phosphate ester nucleotide bonds located between nucleotide positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23; and / or (xii) VP modification at the 5' end.

20. The dsRNA of claim 17, wherein the sense strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at position 9; preferably, wherein the sense strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 9 and 11; more preferably, wherein the sense strand has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 16, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 7, 9, and 11.

21. The dsRNA of claim 17 or 20, wherein the sense strand comprises: (i) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, and 11; or (ii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 9, 11, and 13; or (iii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 15; or (iv) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21; the 2'-fluoro modified nucleotides at positions 7, 9, and 10; and the deoxyribonucleotides at position 11; or (v) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 13; or (vi) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 16; or (vii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 17; or (viii) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 13, 14, 15, 16, 17, 18, 19, 20, and 21, and the 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 12; or (ix) Starting from the 5' end, count the 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and the 2'-fluoro modified nucleotides at positions 8, 9, 10, 11; or (x) Starting from the 5' end, count the nucleotides with 2'-O-methyl modification at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 15, 16, 17, 18, 19, 20, 21, and the nucleotides with 2'-fluorine modification at positions 9, 11, 14; and / or (xi) Starting from the 5' end, count the internucleotide bonds of phosphate thioesters located between nucleotide positions 1 and 2 and between 20 and 21; and / or (xii) is modified by invAb at the 3' end.

22. The dsRNA of claim 17, wherein: (i) The antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 12, 14, and 16; and thiolated nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 (counted from the 5' end), and 2'-fluoro modified nucleotides at positions 7, 9, and 11, and thiophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21 (counted from the 5' end); or (ii) The antisense strand has a length of 23 nucleotides and contains 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 6, 8, 9, 12, 14, and 16, and sulfur-containing nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 9, 11, and 13, and phosphophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end; or (iii) The antisense strand has a length of 23 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 7, 8, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 9, 12, 14, and 16, and sulfur-containing nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 9, 11, and 13, and phosphophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end; or (iv) The antisense strand has a length of 23 nucleotides and contains 2'-O-methyl modified nucleotides at positions 1, 3, 5, 6, 7, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 4, 12, 14, and 16, and sulfur-containing nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 (counted from the 5' end), and 2'-fluoro modified nucleotides at positions 7, 9, and 11, and phosphophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21 (counted from the 5' end); or (v) The antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 10, 14, and 16; and thiolated nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 15, and thiophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end; or (vi) The antisense strand has a length of 23 nucleotides and contains 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 6, 8, 9, 10, 14, and 16; and phosphate thioester nucleosides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Acid-to-nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 (counted from the 5' end), 2'-fluoro modified nucleotides at positions 7, 9, and 10, and a deoxyribonucleotide at position 11, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21 (counted from the 5' end); or (vii) The antisense strand has a length of 23 nucleotides and contains 2'-O-methyl modified nucleotides at positions 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20; and phosphate thioester nucleosides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Acid-to-nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 (counted from the 5' end), 2'-fluoro modified nucleotides at positions 7, 9, and 10, and a deoxyribonucleotide at position 11, as well as phosphate thioester nucleotide bonds between positions 1 and 2 and between positions 20 and 21 (counted from the 5' end); or (viii) The antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 6, 14, 16, and 18; and nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 14, 15, 16, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 13, and phosphophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end; or (ix) The antisense strand has a length of 23 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 8, 9, 14, and 16, and sulfur-containing nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 17, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 16, and phosphophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end; or (x) The antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 8, 9, 14, and 16; and thiolated nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 18, 19, 20, and 21, counted from the 5' end, and 2'-fluoro modified nucleotides at positions 7, 9, 10, 11, and 17, and thiophosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21, counted from the 5' end; or (xi) The antisense strand has a length of 23 nucleotides and includes 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 7, 14, and 16; and phosphate thioester nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Inter-nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 13, 14, 15, 16, 17, 18, 19, 20, and 21 (counted from the 5' end), 2'-fluorinated modified nucleotides at positions 7, 9, 10, 11, and 12, and an invAb modification at the 3' end, and inter-nucleotide bonds of thiophosphate nucleotides between positions 1 and 2 and between positions 20 and 21 (counted from the 5' end); or (xii) The antisense strand has a length of 23 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 7, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 2, 6, 8, 9, 14, and 16, and sulfur-containing nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Phosphophosphate nucleotide bonds; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 (counted from the 5' end), and 2'-fluoro modified nucleotides at positions 8, 9, 10, 11, and phosphate nucleotide bonds between positions 1 and 2 and between positions 20 and 21 (counted from the 5' end); or (xiii) The antisense strand has a length of 23 nucleotides and contains 2'-O-methyl modified nucleotides at positions 1, 3, 4, 5, 6, 8, 9, 10, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 23, counted from the 5' end; 2'-fluoro modified nucleotides at positions 2, 7, 12, 14, and 16; and phosphate thioester nucleotides at positions 1 and 2, 2 and 3, 21 and 22, and 22 and 23, counted from the 5' end. Inter-nucleotide bonds, and VP modification at the 5' end; and the positive strand has a length of 21 nucleotides and has 2'-O-methyl modified nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 15, 16, 17, 18, 19, 20, 21, counted from the 5' end, and 2'-fluorinated modified nucleotides at positions 9, 11, 14, and inter-nucleotide bonds of thiophosphate nucleotides between nucleotide positions 1 and 2 and between nucleotide positions 20 and 21, counted from the 5' end.

23. The dsRNA of any one of claims 14-22, wherein the antisense strand comprises any one of the antisense strand sequences shown in Table 8.

24. The dsRNA of any one of claims 14-23, wherein the sense strand comprises any one of the sense strand sequences shown in Table 8.

25. The dsRNA of claim 23 or 24, wherein the dsRNA comprises any pair of paired sense and antisense sequences selected from those shown in Table 8.

26. The dsRNA of claim 25, wherein the dsRNA comprises any pair of paired sense and antisense sequences selected from: (1) Justice chain: SEQ ID NO:293, Antisense chain: SEQ ID NO:294; (2) Justice chain: SEQ ID NO:399, Antisense chain: SEQ ID NO:400; (3) Justice chain: SEQ ID NO:479, Antisense chain: SEQ ID NO:480; (4) Justice chain: SEQ ID NO:527, Antisense chain: SEQ ID NO:528; (5) Justice chain: SEQ ID NO:533, Antisense chain: SEQ ID NO:534; (6) Justice chain: SEQ ID NO:595, Antisense chain: SEQ ID NO:596; (7) Justice chain: SEQ ID NO:597, Antisense chain: SEQ ID NO:598; (8) Justice chain: SEQ ID NO:607, Antisense chain: SEQ ID NO:608; (9) Justice chain: SEQ ID NO:617, Antisense chain: SEQ ID NO:618; (10) Justice chain: SEQ ID NO:627, Antisense chain: SEQ ID NO:628; (11) Justice chain: SEQ ID NO:687, Antisense chain: SEQ ID NO:688; (12) Justice chain: SEQ ID NO:693, Antisense chain: SEQ ID NO:694; (13) Justice chain: SEQ ID NO:697, Antisense chain: SEQ ID NO:698; (14) Justice chain: SEQ ID NO:739, Antisense chain: SEQ ID NO:740; (15) Justice chain: SEQ ID NO:787, Antisense chain: SEQ ID NO:788; (16) Justice chain: SEQ ID NO:797, Antisense chain: SEQ ID NO:798; (17) Justice chain: SEQ ID NO:801, Antisense chain: SEQ ID NO:

802.

27. The dsRNA of any one of claims 1-26, wherein the dsRNA is further conjugated to a target ligand, preferably the positive strand is conjugated to the target ligand, more preferably the 5' end of the positive strand is conjugated to the target ligand.

28. The dsRNA of claim 27, wherein the targeting ligand is an integrin targeting ligand, preferably an αvβ6 integrin targeting ligand.

29. The dsRNA of claim 28, wherein the targeting ligand has the following structure: in This indicates the position where the dsRNA is attached to the positive strand via a phosphate group or a thiophosphate group.

30. The dsRNA of claim 29, wherein the dsRNA comprises any pair of paired sense and antisense strand sequences selected from the following, and the 5' end of the sense strand is conjugated to the ligand Tri-SM6.1 via a phosphate thioester or phosphodiester bond: (8) Justice chain: SEQ ID NO:399, Antisense chain: SEQ ID NO:400; (9) Justice chain: SEQ ID NO:479, Antisense chain: SEQ ID NO:480; (10) Justice chain: SEQ ID NO:533, Antisense chain: SEQ ID NO:534; (11) Justice chain: SEQ ID NO:597, Antisense chain: SEQ ID NO:598; (12) Justice chain: SEQ ID NO:607, Antisense chain: SEQ ID NO:608; (13) Justice chain: SEQ ID NO:617, Antisense chain: SEQ ID NO:618; (14) Justice chain: SEQ ID NO:627, Antisense chain: SEQ ID NO:

628.

31. A vector comprising a nucleotide sequence encoding the dsRNA of any one of claims 1-30.

32. A cell comprising the dsRNA of any one of claims 1-30 or the vector of claim 31.

33. A pharmaceutical composition comprising any one of claims 1-30, the vector of claim 31, or the cell of claim 32, and optionally a pharmaceutically acceptable carrier or excipient.

34. A kit comprising dsRNA of any one of claims 1-30, the vector of claim 31, or the cell of claim 32.

35. A method for inhibiting the expression of the MUC5AC gene in cells, the method comprising contacting the cells with dsRNA according to any one of claims 1-30 or the pharmaceutical composition according to claim 33.

36. A method for inhibiting the expression of the MUC5AC gene in the cells of a subject, the method comprising administering to the subject any of the dsRNA of any one of claims 1-30, the vector of claim 31, the cells of claim 32, or the pharmaceutical composition of claim 33.

37. A method for treating a subject who benefits from reduced MUC5AC gene expression, the method comprising administering to the subject an effective amount of any one of claims 1-30, the vector of claim 31, the cell of claim 32, or the pharmaceutical composition of claim 33.

38. The method of claim 37, wherein the disease or condition in which the patient benefits from reduced MUC5AC gene expression is a disease mediated or associated with MUC5AC expression.

39. The method of claim 38, wherein the disease mediated or associated with MUC5AC expression is a respiratory disease.

40. The method of claim 38, wherein the disease mediated or associated with MUC5AC expression is a mucus obstructive airway disease, preferably selected from asthma, COPD, and bronchiectasis.

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