Double-stranded RNA that inhibits the production of hepatitis B virus proteins, and pharmaceutical composition

Abstract

The present invention addresses the problem of providing a double-stranded RNA that is useful as a novel anti-hepatitis B virus agent and a pharmaceutical composition that includes the double-stranded RNA. According to the present invention, provided is a double-stranded RNA that inhibits production of a hepatitis B virus protein, wherein the double-stranded RNA inhibits the expression of YTHDC1.

Description

[Technical field]

[0001] The present invention relates to a double-stranded RNA having anti-hepatitis B virus activity. [Background technology]

[0002] Hepatitis B virus (HBV) is a spherical DNA virus that belongs to the Hepadnavirus genus and is approximately 42 nm in diameter. The virus itself is a DNA virus consisting of an envelope, an incomplete double-stranded DNA, a core of 27 nm in diameter that contains DNA polymerase and reverse transcriptase, and is called a Dane particle. It forms a double structure, with the outside consisting of HBs antigen (hepatitis B surface antigen, HBsAg) and the inside consisting of HBc antigen (hepatitis B core antigen, HBcAg) and genetic DNA (Non-Patent Document 1). In addition to Dane particles, HBs antigen exists as hollow particles, small spherical particles, or rod-shaped particles. The HBV genomic DNA has four types of open reading frames (ORFs): the pre-S / S gene region, the P gene region, the X gene region, and the pre-C / C gene region. The core promoter of the X gene region and the precore region of the C gene region are involved in the production of the HBe antigen (hepatitis B envelope antigen, HBeAg) component protein, and point mutations in this region prevent the production of the HBe antigen component protein, resulting in HBe antigen negativity. When HBV invades hepatocytes, the incomplete circular double-stranded DNA is incorporated into the nucleus of the hepatocyte and converted into covalently closed circular DNA (cccDNA). In a chronically infected state of HBV, 5-50 cccDNAs exist as minichromosomes per hepatocyte. Four types of mRNA are transcribed from the cccDNA in the hepatocyte nucleus, and structural proteins HBs antigen, HBc antigen, and HBe antigen, X protein, and polymerase including reverse transcriptase are translated from them. One of the mRNAs is incorporated into the core particle as pregenomic RNA, and a minus-strand DNA is synthesized by the action of reverse transcriptase, followed by a plus-strand DNA to become an incomplete circular double-stranded DNA. Furthermore, it is wrapped in an envelope formed by HBs antigen to become a virus particle (Dane particle), which is released into the blood. Hollow particles (particles without a DNA core) containing HBs antigen, HBc antigen, and p22cr antigen translated by mRNA, and HBe antigen that passes through the liver cell membrane are released and secreted in large quantities into the blood, separate from the route of blood release of Dane particles. This action is thought to be an escape from the attack of the host immune system in HBV infection. HBe antigen, HBc antigen, and p22cr antigen can be measured as HBcr antigen (hepatitis B core-related antigen, HBcrAg) and are used as markers of viral replication.

[0003] Hepatitis B is a type of viral hepatitis caused by infection with HBV, and the number of infected people worldwide is said to be about 240 million. As mentioned above, when infected with HBV, Dane particles are released into the blood, and a large amount of viral proteins are secreted into the blood separately from the route of Dane particle release. It has been pointed out that among viral proteins, HBs antigen in particular may interfere with the elimination of infected cells by the immune system of the host human (Non-Patent Documents 2 and 3). Therefore, it is considered important to treat hepatitis B by reducing Dane particles, that is, by suppressing the proliferation of the virus and reducing viral proteins including HBs antigen. Nucleic acid analogs such as entecavir and tenofovir are used as first-line drugs for hepatitis B. Nucleic acid analog drugs inhibit the activity of HBV polymerase protein and inhibit the production of incomplete circular double-stranded DNA, thereby reducing Dane particles in the blood, i.e., suppressing viral proliferation. However, since nucleic acid analog drugs cannot reduce HBs antigen, it is difficult to achieve elimination of infected cells by the host immune mechanism. Therefore, the development of drugs that can reduce HBs antigen is desired.

[0004] The YT-521B homology (YTH) domain-containing protein family is a group of proteins consisting of YTHDC1, YTHDC2, YTHDF1, YTHDF2, and YTHDF3. It has been revealed that all of these proteins bind to RNA via a recognition sequence containing methylated adenosine (m6A) on the RNA, and control vital phenomena such as RNA splicing and translation into protein (Non-Patent Document 4). It has been disclosed that compounds that focus on the function of YTH family molecules and inhibit them are useful for treating infectious diseases and cancer (Patent Documents 1 and 2). However, Patent Documents 1 and 2 do not contain any description of the relationship between inhibition of YTHDC1 and anti-HBV activity. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Special Publication No. 2018-503663 [Patent Document 2] International Publication No. 2010 / 134939 Brochure [Non-patent literature]

[0006] [Non-Patent Document 1] Tanaka et al., Modern Media, Vol. 54, No. 12, pp. 347-352. [Non-Patent Document 2] Brouw et al., Immunology, vol. 126, pp. 280-289, 2008 [Non-Patent Document 3] Vanlandschoot et al., Journal of General Virology, Vol. 83, pp. 1281-1289, 2002 [Non-Patent Document 4] Liao et al. (2018) Genomics Proteomics Bioinformatics, Volume 16, Pages 99-107 Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a double-stranded RNA having anti-hepatitis B virus (HBV) activity and a pharmaceutical composition comprising the double-stranded RNA. Another object of the present invention is to provide a double-stranded RNA that inhibits the production of hepatitis B virus proteins and a pharmaceutical composition comprising the double-stranded RNA. [Means for solving the problem]

[0008] As a result of extensive research into the above-mentioned problems, the inventors discovered that a specific double-stranded RNA that inhibits the expression of YTHDC1 has the ability to inhibit the production of HBV proteins, thereby completing the present invention.

[0009] That is, the present invention provides the following. <1> A double-stranded RNA that inhibits the production of Hepatitis B virus proteins (HBV proteins), the double-stranded RNA being formed from a sense strand and an antisense strand, the sense strand being selected from the group consisting of SEQ ID NOs: 1, 7, 11, 13, 17, 19, 25, 27, 29, 31, 33, 35, 37, 76, 80, 84, 86, 88, 92, 96, 102, 104, 110, 112, 114, 118, 12 8, 132, 136, 138, 140, 142, 144, 150, 152, 158, 160, 164, 168, 174, 176, 178, 182, 184, 186, 188, 194, 196, 198, 200, 202, 204, 206, 210, 212, 214, 220, 222, 226, 234, 236, 238, 242, 244, 246, 248, 250, 252, 254 , 256, 258, 260, 262, 264, 266, 268, 270, 280, 282, 284, 286, 288, 290, 292, 294, 296, 302, 308, 312, 316, 322, 330, 332, 336, 344, 346, 348, 350, 352, 356, 358, 360, 364, 368, 372, 378, 380, 386, 390, 392, 394, A double-stranded RNA comprising a nucleotide sequence of any one of 396, 398, 400, 402, 404, 406, 410, 412, 414, 416, 418, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, and 446, or comprising a nucleotide sequence having 90% or more sequence identity with the above nucleotide sequence. <2> The antisense strand is selected from SEQ ID NOs: 2, 8, 12, 14, 18, 20, 26, 28, 30, 32, 34, 36, 38, 77, 81, 85, 87, 89, 93, 97, 103, 105, 111, 113, 115, 119, 129, 133, 137, 139, 141, 143, 145, 151, 153, 159, 161, 165, 169, 17 5, 177, 179, 183, 185, 187, 189, 195, 197, 199, 201, 203, 205, 207, 211, 213, 215, 221, 223, 227, 235, 237, 239, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271 , 281, 283, 285, 287, 289, 291, 293, 295, 297, 303, 309, 313, 317, 323, 331, 333, 337, 345, 347, 349, 351, 353, 357, 359, 361, 365, 369, 373, 379, 381, 387, 391, 393, 395, 397, 399, 401, Contains a nucleotide sequence of 403, 405, 407, 411, 413, 415, 417, 419, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, or 447, or contains a nucleotide sequence having 90% or more sequence identity with the above nucleotide sequence, <1> The double-stranded RNA according to claim 1. <3> The combination of the sense strand and the antisense strand is selected from the following: <1> or <2> any one of the double-stranded RNAs described above; (a) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:1 and an antisense strand containing the nucleotide sequence of SEQ ID NO:2; (b) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 7 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 8; (c) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 11 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 12. (d) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 13 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 14; (e) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 17 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 18; (f) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 19 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 20; (g) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 25 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 26. (h) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 27 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 28. (i) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 29 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 30; (j) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 31 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 32. (k) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 33 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 34. (l) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 35 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 36; (m) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:37 and an antisense strand containing the nucleotide sequence of SEQ ID NO:38. (n) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 76 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 77. (o) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 80 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 81. (p) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 84 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 85. (q) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:86 and an antisense strand containing the nucleotide sequence of SEQ ID NO:87. (r) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:88 and an antisense strand containing the nucleotide sequence of SEQ ID NO:89. (s) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:92 and an antisense strand containing the nucleotide sequence of SEQ ID NO:93. (t) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:96 and an antisense strand containing the nucleotide sequence of SEQ ID NO:97. (u) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 102 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 103. (v) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:104 and an antisense strand containing the nucleotide sequence of SEQ ID NO:105. (w) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:110 and an antisense strand containing the nucleotide sequence of SEQ ID NO:111. (x) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:112 and an antisense strand containing the nucleotide sequence of SEQ ID NO:113. (y) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:114 and an antisense strand containing the nucleotide sequence of SEQ ID NO:115. (z) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:118 and an antisense strand containing the nucleotide sequence of SEQ ID NO:119. (aa) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:128 and an antisense strand containing the nucleotide sequence of SEQ ID NO:129. (ab) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 132 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 133. (ac) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 136 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 137. (ad) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:138 and an antisense strand containing the nucleotide sequence of SEQ ID NO:139. (ae) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:140 and an antisense strand containing the nucleotide sequence of SEQ ID NO:141. (af) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 142 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 143. (ag) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:144 and an antisense strand containing the nucleotide sequence of SEQ ID NO:145. (ah) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:150 and an antisense strand containing the nucleotide sequence of SEQ ID NO:151. (ai) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 152 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 153. (aj) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:158 and an antisense strand containing the nucleotide sequence of SEQ ID NO:159. (ak) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:160 and an antisense strand containing the nucleotide sequence of SEQ ID NO:161. (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (am) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:168 and an antisense strand containing the nucleotide sequence of SEQ ID NO:169. (an) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 174 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 175. (ao) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:176 and an antisense strand containing the nucleotide sequence of SEQ ID NO:177. (ap) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:178 and an antisense strand containing the nucleotide sequence of SEQ ID NO:179. (aq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:182 and an antisense strand containing the nucleotide sequence of SEQ ID NO:183. (ar) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 184 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 185. (as) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 186 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 187. (at) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:188 and an antisense strand containing the nucleotide sequence of SEQ ID NO:189. (au) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:194 and an antisense strand containing the nucleotide sequence of SEQ ID NO:195. (av) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:196 and an antisense strand containing the nucleotide sequence of SEQ ID NO:197. (aw) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:198 and an antisense strand containing the nucleotide sequence of SEQ ID NO:199. (ax) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 200 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 201. (ay) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 202 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 203. (az) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 204 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 205. (ba) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:206 and an antisense strand containing the nucleotide sequence of SEQ ID NO:207. (bb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:210 and an antisense strand containing the nucleotide sequence of SEQ ID NO:211. (bc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 212 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 213. (bd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:214 and an antisense strand containing the nucleotide sequence of SEQ ID NO:215. (be) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 220 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 221. (bf) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:222 and an antisense strand containing the nucleotide sequence of SEQ ID NO:223. (bg) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:226 and an antisense strand containing the nucleotide sequence of SEQ ID NO:227. (bh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:234 ​​and an antisense strand containing the nucleotide sequence of SEQ ID NO:235. (bi) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 236 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 237. (bj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:238 and an antisense strand containing the nucleotide sequence of SEQ ID NO:239. (bk) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:242 and an antisense strand containing the nucleotide sequence of SEQ ID NO:243. (bl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:244 and an antisense strand containing the nucleotide sequence of SEQ ID NO:245. (bm) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:246 and an antisense strand containing the nucleotide sequence of SEQ ID NO:247. (bn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:248 and an antisense strand containing the nucleotide sequence of SEQ ID NO:249. (bo) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:250 and an antisense strand containing the nucleotide sequence of SEQ ID NO:251. (bp) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:252 and an antisense strand containing the nucleotide sequence of SEQ ID NO:253. (bq) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:254 and an antisense strand containing the nucleotide sequence of SEQ ID NO:255. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (bs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:258 and an antisense strand containing the nucleotide sequence of SEQ ID NO:259. (bt) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:260 and an antisense strand containing the nucleotide sequence of SEQ ID NO:261. (bu) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:262 and an antisense strand containing the nucleotide sequence of SEQ ID NO:263. (bv) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 264 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 265. (bw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:266 and an antisense strand containing the nucleotide sequence of SEQ ID NO:267. (bx) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:268 and an antisense strand containing the nucleotide sequence of SEQ ID NO:269. (by) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 270 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 271. (bz) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:280 and an antisense strand containing the nucleotide sequence of SEQ ID NO:281. (ca) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 282 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 283. (cb) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:284 and an antisense strand containing the nucleotide sequence of SEQ ID NO:285. (cc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:286 and an antisense strand containing the nucleotide sequence of SEQ ID NO:287. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (ce) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:290 and an antisense strand containing the nucleotide sequence of SEQ ID NO:291. (cf) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:292 and an antisense strand containing the nucleotide sequence of SEQ ID NO:293. (cg) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:294 and an antisense strand containing the nucleotide sequence of SEQ ID NO:295. (ch) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:296 and an antisense strand containing the nucleotide sequence of SEQ ID NO:297. (ci) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 302 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 303. (cj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:308 and an antisense strand containing the nucleotide sequence of SEQ ID NO:309. (ck) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:312 and an antisense strand containing the nucleotide sequence of SEQ ID NO:313. (cl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 316 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 317. (cm) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:322 and an antisense strand containing the nucleotide sequence of SEQ ID NO:323. (cn) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 330 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 331. (co) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 332 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 333. (cp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:336 and an antisense strand containing the nucleotide sequence of SEQ ID NO:337. (cq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 344 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 345. (cr) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:346 and an antisense strand containing the nucleotide sequence of SEQ ID NO:347. (cs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:348 and an antisense strand containing the nucleotide sequence of SEQ ID NO:349. (ct) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:350 and an antisense strand containing the nucleotide sequence of SEQ ID NO:351. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (cv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:356 and an antisense strand containing the nucleotide sequence of SEQ ID NO:357. (cw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:358 and an antisense strand containing the nucleotide sequence of SEQ ID NO:359. (cx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:360 and an antisense strand containing the nucleotide sequence of SEQ ID NO:361. (cy) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:364 and an antisense strand containing the nucleotide sequence of SEQ ID NO:365. (cz) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:368 and an antisense strand containing the nucleotide sequence of SEQ ID NO:369. (da) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 372 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 373. (db) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:378 and an antisense strand containing the nucleotide sequence of SEQ ID NO:379. (dc) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:380 and an antisense strand containing the nucleotide sequence of SEQ ID NO:381. (dd) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:386 and an antisense strand containing the nucleotide sequence of SEQ ID NO:387. (de) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:390 and an antisense strand containing the nucleotide sequence of SEQ ID NO:391. (df) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:392 and an antisense strand containing the nucleotide sequence of SEQ ID NO:393. (dg) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:394 and an antisense strand containing the nucleotide sequence of SEQ ID NO:395. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (di) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:398 and an antisense strand containing the nucleotide sequence of SEQ ID NO:399. (dj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:400 and an antisense strand containing the nucleotide sequence of SEQ ID NO:401. (dk) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:402 and an antisense strand containing the nucleotide sequence of SEQ ID NO:403. (dl) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 404 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 405. (dm) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:406 and an antisense strand containing the nucleotide sequence of SEQ ID NO:407. (dn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:410 and an antisense strand containing the nucleotide sequence of SEQ ID NO:411. (do) ​​a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:412 and an antisense strand containing the nucleotide sequence of SEQ ID NO:413. (dp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:414 and an antisense strand containing the nucleotide sequence of SEQ ID NO:415. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (dr) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:418 and an antisense strand containing the nucleotide sequence of SEQ ID NO:419. (ds) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:422 and an antisense strand containing the nucleotide sequence of SEQ ID NO:423. (dt) dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:424 and an antisense strand containing the nucleotide sequence of SEQ ID NO:425. (du) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 426 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 427. (dv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:428 and an antisense strand containing the nucleotide sequence of SEQ ID NO:429. (dw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:430 and an antisense strand containing the nucleotide sequence of SEQ ID NO:431. (dx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 432 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 433. (dy) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:434 and an antisense strand containing the nucleotide sequence of SEQ ID NO:435. (dz) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:436 and an antisense strand containing the nucleotide sequence of SEQ ID NO:437. (ea) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO:438 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:439. (eb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:440 and an antisense strand containing the nucleotide sequence of SEQ ID NO:441. (ec) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 442 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 443. (ed) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 444 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 445. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447. <4> containing modified nucleotides, <1> ~ <3> The double-stranded RNA according to any one of the preceding claims. <5> The HBV protein is the HBs antigen (HBsAg) protein; <1> ~ <4> The double-stranded RNA according to any one of the preceding claims. <6> In addition, it reduces the expression of HBV DNA. <1> ~ <5> The double-stranded RNA according to any one of the preceding claims. <7> In addition, it reduces the expression of completely closed double-stranded DNA (cccDNA), <1> ~ <6> The double-stranded RNA according to any one of the preceding claims. <8> The double-stranded RNA is an RNA interference agent. <1> ~ <7> The double-stranded RNA according to any one of the preceding claims. <9> <1> ~ <8> A pharmaceutical composition comprising the double-stranded RNA according to any one of the above items. <10> The double-stranded RNA is encoded by a recombinant expression vector. <9> The pharmaceutical composition described in <11> The recombinant expression vector is an adeno-associated virus vector, a retrovirus vector, an adenovirus vector, or an alphavirus vector; <10> The pharmaceutical composition described in <12> Lipid nanoparticles (LNPs) or N-acetylgalactosamine (GalNAc)-containing <9> The pharmaceutical composition described in <13> <1> ~ <8> A vector comprising the double-stranded RNA according to any one of the above items. <14> The vector is an expression vector, and the expression vector is an adeno-associated virus vector, a retrovirus vector, an adenovirus vector, or an alphavirus vector; <13> The vector described in

[0010] The present invention also provides the following. <1> ~ <8> A method for treating a subject infected with HBV or suffering from a disease associated with HBV infection, comprising administering the double-stranded RNA described in any one of the above to a subject infected with HBV or suffering from a disease associated with HBV infection. The disease associated with HBV infection is hepatitis B, liver cirrhosis, or liver cancer. The treatment method described above. <c> <1> ~ <8> any one of the double-stranded RNAs described in any one of the following: Instructions for treating illnesses related to HBV infection A kit for treating a disease associated with HBV infection, comprising: <d> The method for treating a disease associated with HBV infection comprises administering the double-stranded RNA to a subject infected with HBV or suffering from a disease associated with HBV infection. <c>A kit for treating a disease associated with HBV infection as described above. <e> A double-stranded RNA that inhibits the expression of YTHDC1 for use in a treatment to inhibit the production of Hepatitis B virus proteins (HBV proteins). <f> An inhibitor of YTHDC1 for use in the treatment of hepatitis B. <g> Use of a double-stranded RNA that inhibits the expression of YTHDC1 for the production of a double-stranded RNA that inhibits the production of a hepatitis B virus protein (HBV protein). <h> Use of an inhibitor of YTHDC1 for the production of double-stranded RNA for treating hepatitis B in a subject. Effect of the Invention

[0011] The double-stranded RNA of the present invention has excellent anti-HBV activity and is useful as an anti-HBV agent. [Brief description of the drawings]

[0012] < / h> < / g> < / f> < / e> < / c> < / d> < / c> [Figure 1] FIG. 1 shows the relative cell viability (%) of wells to which each concentration of dsRNA was added, relative to wells to which negative control dsRNA was added. [Diagram 2] FIG. 2 shows the relative values ​​(%) of HBs antigen (HBsAg) in wells to which each concentration of dsRNA was added, relative to wells to which negative control dsRNA was added at each concentration. [Diagram 3] FIG. 3 shows the relative values ​​(%) of HBV DNA in wells to which each concentration of dsRNA was added, relative to the values ​​in wells to which negative control dsRNA was added at each concentration. [Figure 4] FIG. 4 shows the relative values ​​(%) of the YTHDC1 expression level in wells to which each dsRNA was added, relative to the YTHDC1 expression level in wells to which negative control dsRNA was added. [Diagram 5] FIG. 5 shows the relative cell viability (%) of wells to which each concentration of dsRNA was added, relative to wells to which each concentration of negative control dsRNA was added. [Figure 6] FIG. 6 shows the relative values ​​(%) of HBs antigen in wells to which each concentration of dsRNA was added, compared with wells to which negative control dsRNA was added. [Figure 7] FIG. 7 shows the relative values ​​(%) of HBV DNA in wells to which each concentration of dsRNA was added, compared with wells to which each concentration of negative control dsRNA was added. [Figure 8] FIG. 8 shows the relative values ​​(%) of the YTHDC1 expression level in wells to which each dsRNA was added, relative to the YTHDC1 expression level in wells to which negative control dsRNA was added. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The present invention will be described in detail below. In this specification, unless otherwise specified, each term has the following meaning.

[0014] Prevention means inhibiting the onset, reducing the risk of onset, or delaying the onset. Treatment means, for example, amelioration or inhibition of progression of a disease or condition. Treatment means prevention or treatment for various diseases. Effective amount means a therapeutically effective amount or a prophylactically effective amount, etc. A therapeutically effective amount is an amount sufficient for the stabilization of HBV infection, the reduction of HBV infection, or the eradication of HBV infection in an infected subject. Also, a prophylactically effective amount is an amount sufficient for the prevention of HBV infection in a subject at risk of infection. Subject means a mammal including a human.

[0015] <Hepatitis B virus (HBV)> Hepatitis B virus (HBV) means a virus having the ability to cause hepatitis B. HBV is currently classified into eight genotypes (genotype) from type A to type H due to differences in nucleotide sequences derived from genetic mutations. The HBV targeted for prevention or treatment in the double-stranded RNA of the present invention includes all genotypes. In the present invention, diseases associated with HBV infection include hepatitis B, including acute hepatitis, chronic hepatitis, and fulminant hepatitis. Also, any disease caused by HBV infection of a living body including a human also includes liver cancers such as liver cirrhosis, liver fibrosis, and hepatocellular carcinoma.

[0016] <Hepatitis B virus protein (HBV protein)> Hepatitis B virus protein (HBV protein) is a protein constituting HBV, and examples include HBs antigen, HBc antigen, HBe antigen, etc. In the present invention, the HBV protein is not particularly limited, but is preferably the HBs antigen.

[0017] "Double-stranded RNA" includes, but is not limited to, nucleic acids that interfere with or inhibit the expression of a target gene by short interfering RNA (siRNA), small hairpin RNA (shRNA), or RNA interference (RNAi). "RNA interfering agent" is a nucleic acid that interferes with or inhibits the expression of a target gene via RNA interference. RNA interference is a post-transcriptional targeted gene silencing technique that uses double-stranded RNA (dsRNA) to degrade mRNA that contains the same sequence as the dsRNA (Sharp et al., Science, vol. 287, pp. 2431-2432, 2000; Zamore et al., Cell, vol. 101, pp. 25-33, 2000; Tuschl et al., Genes & Development, vol. 13, pp. 3191-3197, 1999). It occurs when endogenous ribonucleases cleave long dsRNA to generate shorter 21 or 22 nucleotide long RNAs called short interfering RNAs (siRNAs). The siRNAs mediate the degradation of the target mRNA. Kits for the synthesis of RNAi are commercially available, for example from New England Biolabs and Ambion. In one embodiment, one or more of the above kits for use with antisense RNA can be used. The double-stranded RNA of the present invention is preferably an RNA interference agent.

[0018] The sense strand of the present invention is SEQ ID NO: 1, 7, 11, 13, 17, 19, 25, 27, 29, 31, 33, 35, 37, 76, 80, 84, 86, 88, 92, 96, 102, 104, 110, 112, 114, 118, 128, 132, 136, 138, 140, 142, 144, 150, 152, 158, 160, 164, 168, 1 74, 176, 178, 182, 184, 186, 188, 194, 196, 198, 200, 202, 204, 206, 210, 212, 214, 220, 222, 226, 234, 236, 238, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270 , 280, 282, 284, 286, 288, 290, 292, 294, 296, 302, 308, 312, 316, 322, 330, 332, 336, 344, 346, 348, 350, 352, 356, 358, 360, 364, 368, 372, 378, 380, 386, 390, 392, 394, 396, 398, 400, It contains a nucleotide sequence of nucleotides represented by any of 402, 404, 406, 410, 412, 414, 416, 418, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, and 446, or contains a nucleotide sequence having 90% or more sequence identity with the above nucleotide sequence. The sense strand of the present invention is SEQ ID NO: 1, 7, 11, 13, 17, 19, 25, 27, 29, 31, 33, 35, 37, 76, 80, 84, 86, 88, 92, 96, 102, 104, 110, 112, 114, 118, 128, 132, 136, 138, 140, 142, 144, 150, 152, 158, 160, 164, 168, 174 , 176, 178, 182, 184, 186, 188, 194, 196, 198, 200, 202, 204, 206, 210, 212, 214, 220, 222, 226, 234, 236, 238, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 28 0, 282, 284, 286, 288, 290, 292, 294, 296, 302, 308, 312, 316, 322, 330, 332, 336, 344, 346, 348, 350, 352, 356, 358, 360, 364, 368, 372, 378, 380, 386, 390, 392, 394, 396, 398, 400, 402, 4 It is preferable that the nucleic acid sequence contains a nucleotide sequence represented by any one of the following: 04, 406, 410, 412, 414, 416, 418, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, or contains a nucleotide sequence having 92% or more sequence identity with the above nucleotide sequence. The sense strand of the present invention is SEQ ID NO: 1, 7, 11, 13, 17, 19, 25, 27, 29, 31, 33, 35, 37, 76, 80, 84, 86, 88, 92, 96, 102, 104, 110, 112, 114, 118, 128, 132, 136, 138, 140, 142, 144, 150, 152, 158, 160, 164, 168, 174 , 176, 178, 182, 184, 186, 188, 194, 196, 198, 200, 202, 204, 206, 210, 212, 214, 220, 222, 226, 234, 236, 238, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 280 , 282, 284, 286, 288, 290, 292, 294, 296, 302, 308, 312, 316, 322, 330, 332, 336, 344, 346, 348, 350, 352, 356, 358, 360, 364, 368, 372, 378, 380, 386, 390, 392, 394, 396, 398, 400, 402, 404 , 406, 410, 412, 414, 416, 418, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, or more preferably, comprises a nucleotide sequence having 95% or more sequence identity with the above nucleotide sequence. The sense strand of the present invention is SEQ ID NO: 1, 7, 11, 13, 17, 19, 25, 27, 29, 31, 33, 35, 37, 76, 80, 84, 86, 88, 92, 96, 102, 104, 110, 112, 114, 118, 128, 132, 136, 138, 140, 142, 144, 150, 152, 158, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 198, 199, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 210, 210, 211, 4, 168, 174, 176, 178, 182, 184, 186, 188, 194, 196, 198, 200, 202, 204, 206, 210, 212, 214, 220, 222, 226, 234, 236, 238, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 280, 282, 284, 286, 288, 290, 292, 294, 296, 302, 308, 312, 316, 322, 330, 332, 336, 344, 346, 348, 350, 352, 356, 358, 360, 364, 368, 372, 378, 380, 386, 39 It is even more preferable that the nucleotide sequence comprises any of the nucleotides 0, 392, 394, 396, 398, 400, 402, 404, 406, 410, 412, 414, 416, 418, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, or 446.

[0019] The antisense strands of the present invention are SEQ ID NOs: 2, 8, 12, 14, 18, 20, 26, 28, 30, 32, 34, 36, 38, 77, 81, 85, 87, 89, 93, 97, 103, 105, 111, 113, 115, 119, 129, 133, 137, 139, 141, 143, 145, 151, 153, 159, 161, 165, 169 , 175, 177, 179, 183, 185, 187, 189, 195, 197, 199, 201, 203, 205, 207, 211, 213, 215, 221, 223, 227, 235, 237, 239, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 2 71, 281, 283, 285, 287, 289, 291, 293, 295, 297, 303, 309, 313, 317, 323, 331, 333, 337, 345, 347, 349, 351, 353, 357, 359, 361, 365, 369, 373, 379, 381, 387, 391, 393, 395, 397, 399, 401 , 403, 405, 407, 411, 413, 415, 417, 419, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, or a nucleotide sequence having 90% or more sequence identity with the above nucleotide sequence. The antisense strands of the present invention are SEQ ID NOs: 2, 8, 12, 14, 18, 20, 26, 28, 30, 32, 34, 36, 38, 77, 81, 85, 87, 89, 93, 97, 103, 105, 111, 113, 115, 119, 129, 133, 137, 139, 141, 143, 145, 151, 153, 159, 161, 165, 169, 170, 172, 174, 176, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 198, 199, 198, 199, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 109, 110, 111, 112, 113, 115, 119, 129, 133, 137, 139, 141, 143, 145, 151, 153, 159, 161, 165, 169, 170, 171, 172, 173, 1 75, 177, 179, 183, 185, 187, 189, 195, 197, 199, 201, 203, 205, 207, 211, 213, 215, 221, 223, 227, 235, 237, 239, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 2 81, 283, 285, 287, 289, 291, 293, 295, 297, 303, 309, 313, 317, 323, 331, 333, 337, 345, 347, 349, 351, 353, 357, 359, 361, 365, 369, 373, 379, 381, 387, 391, 393, 395, 397, 399, 401, 403, 4 It is preferable that the nucleic acid sequence contains a nucleotide sequence represented by any one of the following nucleotides: 05, 407, 411, 413, 415, 417, 419, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, or contains a nucleotide sequence having 92% or more sequence identity with the above nucleotide sequence. The antisense strands of the present invention are SEQ ID NOs: 2, 8, 12, 14, 18, 20, 26, 28, 30, 32, 34, 36, 38, 77, 81, 85, 87, 89, 93, 97, 103, 105, 111, 113, 115, 119, 129, 133, 137, 139, 141, 143, 145, 151, 153, 159, 161, 165, 169, 170, 172, 174, 176, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 198, 199, 198, 199, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 109, 110, 111, 112, 113, 115, 119, 129, 133, 137, 139, 141, 143, 145, 151, 153, 159, 161, 165, 169, 170, 171, 172, 173, 1 75, 177, 179, 183, 185, 187, 189, 195, 197, 199, 201, 203, 205, 207, 211, 213, 215, 221, 223, 227, 235, 237, 239, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 28 1, 283, 285, 287, 289, 291, 293, 295, 297, 303, 309, 313, 317, 323, 331, 333, 337, 345, 347, 349, 351, 353, 357, 359, 361, 365, 369, 373, 379, 381, 387, 391, 393, 395, 397, 399, 401, 403, 40 It is more preferable that the nucleic acid sequence contains a nucleotide sequence represented by any one of the following nucleotides: 5, 407, 411, 413, 415, 417, 419, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, or contains a nucleotide sequence having 95% or more sequence identity with the above nucleotide sequence. The antisense strand of the present invention is SEQ ID NO: 2, 8, 12, 14, 18, 20, 26, 28, 30, 32, 34, 36, 38, 77, 81, 85, 87, 89, 93, 97, 103, 105, 111, 113, 115, 119, 129, 133, 137, 139, 141, 143, 145, 151, 153, 159, 161, 165, 169, 175, 177, 179, 183, 185, 187, 189, 195, 197, 199, 201, 203, 205, 207, 211, 213, 215, 221, 223, 227, 235, 237, 239, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263 , 265, 267, 269, 271, 281, 283, 285, 287, 289, 291, 293, 295, 297, 303, 309, 313, 317, 323, 331, 333, 337, 345, 347, 349, 351, 353, 357, 359, 361, 365, 369, 373, 379, 381, 387, 39 It is even more preferable that the nucleotide sequence comprises any of the nucleotides 1, 393, 395, 397, 399, 401, 403, 405, 407, 411, 413, 415, 417, 419, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, or 447.

[0020] The sense strand and antisense strand of the present invention are 30 nucleotides or less in length, preferably 18 to 25 nucleotides in length, and more preferably 19 to 21 nucleotides in length.

[0021] In the double-stranded RNA of the present invention, the combination of the sense strand and the antisense strand is preferably selected from the following: (a) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:1 and an antisense strand containing the nucleotide sequence of SEQ ID NO:2; (b) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 7 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 8; (c) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 11 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 12. (d) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 13 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 14; (e) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 17 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 18; (f) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 19 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 20; (g) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 25 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 26. (h) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 27 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 28. (i) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 29 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 30; (j) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 31 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 32. (k) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 33 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 34. (l) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 35 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 36; (m) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:37 and an antisense strand containing the nucleotide sequence of SEQ ID NO:38. (n) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 76 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 77. (o) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 80 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 81. (p) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 84 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 85. (q) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:86 and an antisense strand containing the nucleotide sequence of SEQ ID NO:87. (r) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:88 and an antisense strand containing the nucleotide sequence of SEQ ID NO:89. (s) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:92 and an antisense strand containing the nucleotide sequence of SEQ ID NO:93. (t) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:96 and an antisense strand containing the nucleotide sequence of SEQ ID NO:97. (u) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 102 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 103. (v) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:104 and an antisense strand containing the nucleotide sequence of SEQ ID NO:105. (w) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:110 and an antisense strand containing the nucleotide sequence of SEQ ID NO:111. (x) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:112 and an antisense strand containing the nucleotide sequence of SEQ ID NO:113. (y) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:114 and an antisense strand containing the nucleotide sequence of SEQ ID NO:115. (z) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:118 and an antisense strand containing the nucleotide sequence of SEQ ID NO:119. (aa) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:128 and an antisense strand containing the nucleotide sequence of SEQ ID NO:129. (ab) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 132 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 133. (ac) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 136 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 137. (ad) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:138 and an antisense strand containing the nucleotide sequence of SEQ ID NO:139. (ae) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:140 and an antisense strand containing the nucleotide sequence of SEQ ID NO:141. (af) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 142 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 143. (ag) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:144 and an antisense strand containing the nucleotide sequence of SEQ ID NO:145. (ah) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:150 and an antisense strand containing the nucleotide sequence of SEQ ID NO:151. (ai) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 152 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 153. (aj) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:158 and an antisense strand containing the nucleotide sequence of SEQ ID NO:159. (ak) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:160 and an antisense strand containing the nucleotide sequence of SEQ ID NO:161. (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (am) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:168 and an antisense strand containing the nucleotide sequence of SEQ ID NO:169. (an) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 174 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 175. (ao) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:176 and an antisense strand containing the nucleotide sequence of SEQ ID NO:177. (ap) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:178 and an antisense strand containing the nucleotide sequence of SEQ ID NO:179. (aq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:182 and an antisense strand containing the nucleotide sequence of SEQ ID NO:183. (ar) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 184 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 185. (as) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 186 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 187. (at) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:188 and an antisense strand containing the nucleotide sequence of SEQ ID NO:189. (au) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:194 and an antisense strand containing the nucleotide sequence of SEQ ID NO:195. (av) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:196 and an antisense strand containing the nucleotide sequence of SEQ ID NO:197. (aw) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:198 and an antisense strand containing the nucleotide sequence of SEQ ID NO:199. (ax) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 200 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 201. (ay) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 202 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 203. (az) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 204 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 205. (ba) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:206 and an antisense strand containing the nucleotide sequence of SEQ ID NO:207. (bb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:210 and an antisense strand containing the nucleotide sequence of SEQ ID NO:211. (bc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 212 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 213. (bd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:214 and an antisense strand containing the nucleotide sequence of SEQ ID NO:215. (be) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 220 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 221. (bf) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:222 and an antisense strand containing the nucleotide sequence of SEQ ID NO:223. (bg) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:226 and an antisense strand containing the nucleotide sequence of SEQ ID NO:227. (bh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:234 ​​and an antisense strand containing the nucleotide sequence of SEQ ID NO:235. (bi) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 236 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 237. (bj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:238 and an antisense strand containing the nucleotide sequence of SEQ ID NO:239. (bk) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:242 and an antisense strand containing the nucleotide sequence of SEQ ID NO:243. (bl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:244 and an antisense strand containing the nucleotide sequence of SEQ ID NO:245. (bm) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:246 and an antisense strand containing the nucleotide sequence of SEQ ID NO:247. (bn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:248 and an antisense strand containing the nucleotide sequence of SEQ ID NO:249. (bo) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:250 and an antisense strand containing the nucleotide sequence of SEQ ID NO:251. (bp) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:252 and an antisense strand containing the nucleotide sequence of SEQ ID NO:253. (bq) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:254 and an antisense strand containing the nucleotide sequence of SEQ ID NO:255. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (bs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:258 and an antisense strand containing the nucleotide sequence of SEQ ID NO:259. (bt) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:260 and an antisense strand containing the nucleotide sequence of SEQ ID NO:261. (bu) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:262 and an antisense strand containing the nucleotide sequence of SEQ ID NO:263. (bv) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 264 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 265. (bw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:266 and an antisense strand containing the nucleotide sequence of SEQ ID NO:267. (bx) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:268 and an antisense strand containing the nucleotide sequence of SEQ ID NO:269. (by) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 270 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 271. (bz) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:280 and an antisense strand containing the nucleotide sequence of SEQ ID NO:281. (ca) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 282 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 283. (cb) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:284 and an antisense strand containing the nucleotide sequence of SEQ ID NO:285. (cc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:286 and an antisense strand containing the nucleotide sequence of SEQ ID NO:287. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (ce) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:290 and an antisense strand containing the nucleotide sequence of SEQ ID NO:291. (cf) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:292 and an antisense strand containing the nucleotide sequence of SEQ ID NO:293. (cg) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:294 and an antisense strand containing the nucleotide sequence of SEQ ID NO:295. (ch) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:296 and an antisense strand containing the nucleotide sequence of SEQ ID NO:297. (ci) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 302 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 303. (cj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:308 and an antisense strand containing the nucleotide sequence of SEQ ID NO:309. (ck) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:312 and an antisense strand containing the nucleotide sequence of SEQ ID NO:313. (cl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 316 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 317. (cm) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:322 and an antisense strand containing the nucleotide sequence of SEQ ID NO:323. (cn) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 330 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 331. (co) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 332 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 333. (cp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:336 and an antisense strand containing the nucleotide sequence of SEQ ID NO:337. (cq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 344 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 345. (cr) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:346 and an antisense strand containing the nucleotide sequence of SEQ ID NO:347. (cs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:348 and an antisense strand containing the nucleotide sequence of SEQ ID NO:349. (ct) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:350 and an antisense strand containing the nucleotide sequence of SEQ ID NO:351. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (cv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:356 and an antisense strand containing the nucleotide sequence of SEQ ID NO:357. (cw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:358 and an antisense strand containing the nucleotide sequence of SEQ ID NO:359. (cx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:360 and an antisense strand containing the nucleotide sequence of SEQ ID NO:361. (cy) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:364 and an antisense strand containing the nucleotide sequence of SEQ ID NO:365. (cz) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:368 and an antisense strand containing the nucleotide sequence of SEQ ID NO:369. (da) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 372 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 373. (db) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:378 and an antisense strand containing the nucleotide sequence of SEQ ID NO:379. (dc) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:380 and an antisense strand containing the nucleotide sequence of SEQ ID NO:381. (dd) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:386 and an antisense strand containing the nucleotide sequence of SEQ ID NO:387. (de) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:390 and an antisense strand containing the nucleotide sequence of SEQ ID NO:391. (df) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:392 and an antisense strand containing the nucleotide sequence of SEQ ID NO:393. (dg) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:394 and an antisense strand containing the nucleotide sequence of SEQ ID NO:395. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (di) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:398 and an antisense strand containing the nucleotide sequence of SEQ ID NO:399. (dj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:400 and an antisense strand containing the nucleotide sequence of SEQ ID NO:401. (dk) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:402 and an antisense strand containing the nucleotide sequence of SEQ ID NO:403. (dl) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 404 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 405. (dm) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:406 and an antisense strand containing the nucleotide sequence of SEQ ID NO:407. (dn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:410 and an antisense strand containing the nucleotide sequence of SEQ ID NO:411. (do) ​​a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:412 and an antisense strand containing the nucleotide sequence of SEQ ID NO:413. (dp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:414 and an antisense strand containing the nucleotide sequence of SEQ ID NO:415. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (dr) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:418 and an antisense strand containing the nucleotide sequence of SEQ ID NO:419. (ds) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:422 and an antisense strand containing the nucleotide sequence of SEQ ID NO:423. (dt) dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:424 and an antisense strand containing the nucleotide sequence of SEQ ID NO:425. (du) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 426 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 427. (dv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:428 and an antisense strand containing the nucleotide sequence of SEQ ID NO:429. (dw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:430 and an antisense strand containing the nucleotide sequence of SEQ ID NO:431. (dx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 432 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 433. (dy) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:434 and an antisense strand containing the nucleotide sequence of SEQ ID NO:435. (dz) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:436 and an antisense strand containing the nucleotide sequence of SEQ ID NO:437. (ea) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO:438 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:439. (eb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:440 and an antisense strand containing the nucleotide sequence of SEQ ID NO:441. (ec) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 442 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 443. (ed) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 444 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 445. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447.

[0022] In the double-stranded RNA of the present invention, the combination of the sense strand and the antisense strand is more preferably selected from the following double-stranded RNAs: (a) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:1 and an antisense strand containing the nucleotide sequence of SEQ ID NO:2; (b) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 7 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 8; (e) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 17 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 18; (g) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 25 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 26. (h) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 27 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 28. (j) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 31 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 32. (k) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:33 and an antisense strand containing the nucleotide sequence of SEQ ID NO:34. (n) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 76 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 77. (p) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 84 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 85. (q) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:86 and an antisense strand containing the nucleotide sequence of SEQ ID NO:87. (r) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:88 and an antisense strand containing the nucleotide sequence of SEQ ID NO:89. (s) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:92 and an antisense strand containing the nucleotide sequence of SEQ ID NO:93. (y) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:114 and an antisense strand containing the nucleotide sequence of SEQ ID NO:115. (z) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:118 and an antisense strand containing the nucleotide sequence of SEQ ID NO:119. (ab) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 132 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 133. (ad) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:138 and an antisense strand containing the nucleotide sequence of SEQ ID NO:139. (ag) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:144 and an antisense strand containing the nucleotide sequence of SEQ ID NO:145. (aj) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:158 and an antisense strand containing the nucleotide sequence of SEQ ID NO:159. (ak) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:160 and an antisense strand containing the nucleotide sequence of SEQ ID NO:161. (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (am) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:168 and an antisense strand containing the nucleotide sequence of SEQ ID NO:169. (an) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 174 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 175. (ap) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:178 and an antisense strand containing the nucleotide sequence of SEQ ID NO:179. (aq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:182 and an antisense strand containing the nucleotide sequence of SEQ ID NO:183. (ar) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 184 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 185. (as) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 186 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 187. (au) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:194 and an antisense strand containing the nucleotide sequence of SEQ ID NO:195. (aw) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:198 and an antisense strand containing the nucleotide sequence of SEQ ID NO:199. (ax) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 200 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 201. (ay) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 202 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 203. (bb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:210 and an antisense strand containing the nucleotide sequence of SEQ ID NO:211. (bc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 212 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 213. (bd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:214 and an antisense strand containing the nucleotide sequence of SEQ ID NO:215. (be) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 220 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 221. (bf) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:222 and an antisense strand containing the nucleotide sequence of SEQ ID NO:223. (bh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:234 ​​and an antisense strand containing the nucleotide sequence of SEQ ID NO:235. (bk) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:242 and an antisense strand containing the nucleotide sequence of SEQ ID NO:243. (bl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:244 and an antisense strand containing the nucleotide sequence of SEQ ID NO:245. (bn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:248 and an antisense strand containing the nucleotide sequence of SEQ ID NO:249. (bo) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:250 and an antisense strand containing the nucleotide sequence of SEQ ID NO:251. (bp) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:252 and an antisense strand containing the nucleotide sequence of SEQ ID NO:253. (bq) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:254 and an antisense strand containing the nucleotide sequence of SEQ ID NO:255. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (bs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:258 and an antisense strand containing the nucleotide sequence of SEQ ID NO:259. (bt) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:260 and an antisense strand containing the nucleotide sequence of SEQ ID NO:261. (bu) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:262 and an antisense strand containing the nucleotide sequence of SEQ ID NO:263. (bv) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 264 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 265. (bz) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:280 and an antisense strand containing the nucleotide sequence of SEQ ID NO:281. (ca) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 282 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 283. (cb) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:284 and an antisense strand containing the nucleotide sequence of SEQ ID NO:285. (cc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:286 and an antisense strand containing the nucleotide sequence of SEQ ID NO:287. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (ce) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:290 and an antisense strand containing the nucleotide sequence of SEQ ID NO:291. (cg) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:294 and an antisense strand containing the nucleotide sequence of SEQ ID NO:295. (ch) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:296 and an antisense strand containing the nucleotide sequence of SEQ ID NO:297. (ci) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 302 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 303. (cj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:308 and an antisense strand containing the nucleotide sequence of SEQ ID NO:309. (cl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 316 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 317. (cq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 344 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 345. (cs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:348 and an antisense strand containing the nucleotide sequence of SEQ ID NO:349. (ct) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:350 and an antisense strand containing the nucleotide sequence of SEQ ID NO:351. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (cv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:356 and an antisense strand containing the nucleotide sequence of SEQ ID NO:357. (cw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:358 and an antisense strand containing the nucleotide sequence of SEQ ID NO:359. (cx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:360 and an antisense strand containing the nucleotide sequence of SEQ ID NO:361. (cy) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:364 and an antisense strand containing the nucleotide sequence of SEQ ID NO:365. (da) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 372 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 373. (db) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:378 and an antisense strand containing the nucleotide sequence of SEQ ID NO:379. (dc) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:380 and an antisense strand containing the nucleotide sequence of SEQ ID NO:381. (de) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:390 and an antisense strand containing the nucleotide sequence of SEQ ID NO:391. (dg) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:394 and an antisense strand containing the nucleotide sequence of SEQ ID NO:395. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (di) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:398 and an antisense strand containing the nucleotide sequence of SEQ ID NO:399. (dj) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:400 and an antisense strand containing the nucleotide sequence of SEQ ID NO:401. (dl) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 404 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 405. (dm) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:406 and an antisense strand containing the nucleotide sequence of SEQ ID NO:407. (dn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:410 and an antisense strand containing the nucleotide sequence of SEQ ID NO:411. (do) ​​a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:412 and an antisense strand containing the nucleotide sequence of SEQ ID NO:413. (dp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:414 and an antisense strand containing the nucleotide sequence of SEQ ID NO:415. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (dr) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:418 and an antisense strand containing the nucleotide sequence of SEQ ID NO:419. (dt) dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:424 and an antisense strand containing the nucleotide sequence of SEQ ID NO:425. (du) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 426 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 427. (dv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:428 and an antisense strand containing the nucleotide sequence of SEQ ID NO:429. (dw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:430 and an antisense strand containing the nucleotide sequence of SEQ ID NO:431. (dx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 432 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 433. (dy) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:434 and an antisense strand containing the nucleotide sequence of SEQ ID NO:435. (dz) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:436 and an antisense strand containing the nucleotide sequence of SEQ ID NO:437. (ea) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO:438 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:439. (eb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:440 and an antisense strand containing the nucleotide sequence of SEQ ID NO:441. (ec) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 442 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 443. (ed) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 444 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 445. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447.

[0023] In the double-stranded RNA of the present invention, the combination of the sense strand and the antisense strand is more preferably selected from the following double-stranded RNAs: (y) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:114 and an antisense strand containing the nucleotide sequence of SEQ ID NO:115. (z) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:118 and an antisense strand containing the nucleotide sequence of SEQ ID NO:119. (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (ap) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:178 and an antisense strand containing the nucleotide sequence of SEQ ID NO:179. (ar) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 184 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 185. (au) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:194 and an antisense strand containing the nucleotide sequence of SEQ ID NO:195. (bb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:210 and an antisense strand containing the nucleotide sequence of SEQ ID NO:211. (bc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 212 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 213. (bf) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:222 and an antisense strand containing the nucleotide sequence of SEQ ID NO:223. (bl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:244 and an antisense strand containing the nucleotide sequence of SEQ ID NO:245. (bn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:248 and an antisense strand containing the nucleotide sequence of SEQ ID NO:249. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (bs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:258 and an antisense strand containing the nucleotide sequence of SEQ ID NO:259. (bv) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 264 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 265. (ca) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 282 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 283. (cc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:286 and an antisense strand containing the nucleotide sequence of SEQ ID NO:287. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (ce) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:290 and an antisense strand containing the nucleotide sequence of SEQ ID NO:291. (cg) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:294 and an antisense strand containing the nucleotide sequence of SEQ ID NO:295. (ci) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 302 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 303. (cl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 316 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 317. (cq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 344 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 345. (cs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:348 and an antisense strand containing the nucleotide sequence of SEQ ID NO:349. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (cv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:356 and an antisense strand containing the nucleotide sequence of SEQ ID NO:357. (cw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:358 and an antisense strand containing the nucleotide sequence of SEQ ID NO:359. (cx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:360 and an antisense strand containing the nucleotide sequence of SEQ ID NO:361. (dg) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:394 and an antisense strand containing the nucleotide sequence of SEQ ID NO:395. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (dn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:410 and an antisense strand containing the nucleotide sequence of SEQ ID NO:411. (dp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:414 and an antisense strand containing the nucleotide sequence of SEQ ID NO:415. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (du) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 426 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 427. (dv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:428 and an antisense strand containing the nucleotide sequence of SEQ ID NO:429. (ed) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 444 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 445. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447.

[0024] In the double-stranded RNA of the present invention, the combination of the sense strand and the antisense strand is more preferably selected from the following: (y) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:114 and an antisense strand containing the nucleotide sequence of SEQ ID NO:115. (z) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:118 and an antisense strand containing the nucleotide sequence of SEQ ID NO:119. (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (ap) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:178 and an antisense strand containing the nucleotide sequence of SEQ ID NO:179. (ar) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 184 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 185. (au) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:194 and an antisense strand containing the nucleotide sequence of SEQ ID NO:195. (bb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:210 and an antisense strand containing the nucleotide sequence of SEQ ID NO:211. (bc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 212 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 213. (bl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:244 and an antisense strand containing the nucleotide sequence of SEQ ID NO:245. (bn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:248 and an antisense strand containing the nucleotide sequence of SEQ ID NO:249. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (bs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:258 and an antisense strand containing the nucleotide sequence of SEQ ID NO:259. (bv) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 264 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 265. (ca) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 282 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 283. (cc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:286 and an antisense strand containing the nucleotide sequence of SEQ ID NO:287. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (cg) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:294 and an antisense strand containing the nucleotide sequence of SEQ ID NO:295. (ci) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 302 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 303. (cl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 316 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 317. (cs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:348 and an antisense strand containing the nucleotide sequence of SEQ ID NO:349. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (cv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:356 and an antisense strand containing the nucleotide sequence of SEQ ID NO:357. (cw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:358 and an antisense strand containing the nucleotide sequence of SEQ ID NO:359. (cx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:360 and an antisense strand containing the nucleotide sequence of SEQ ID NO:361. (dg) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:394 and an antisense strand containing the nucleotide sequence of SEQ ID NO:395. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (dn) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:410 and an antisense strand containing the nucleotide sequence of SEQ ID NO:411. (dp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:414 and an antisense strand containing the nucleotide sequence of SEQ ID NO:415. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (du) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 426 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 427. (dv) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:428 and an antisense strand containing the nucleotide sequence of SEQ ID NO:429. (ed) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 444 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 445. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447.

[0025] In the double-stranded RNA of the present invention, a double-stranded RNA in which the combination of the sense strand and the antisense strand is selected from the following is particularly preferred. (y) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:114 and an antisense strand containing the nucleotide sequence of SEQ ID NO:115. (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (ar) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 184 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 185. (bb) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:210 and an antisense strand containing the nucleotide sequence of SEQ ID NO:211. (bc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 212 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 213. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (cc) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:286 and an antisense strand containing the nucleotide sequence of SEQ ID NO:287. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (cl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 316 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 317. (cs) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:348 and an antisense strand containing the nucleotide sequence of SEQ ID NO:349. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (cw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:358 and an antisense strand containing the nucleotide sequence of SEQ ID NO:359. (cx) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:360 and an antisense strand containing the nucleotide sequence of SEQ ID NO:361. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (dp) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:414 and an antisense strand containing the nucleotide sequence of SEQ ID NO:415. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447.

[0026] In another embodiment, in the double-stranded RNA of the present invention, a combination of a sense strand and an antisense strand selected from the following double-stranded RNAs is more preferable. (a) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:1 and an antisense strand containing the nucleotide sequence of SEQ ID NO:2; (b) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 7 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 8; (c) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 11 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 12. (e) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 17 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 18; (f) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 19 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 20; (g) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 25 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 26. (h) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 27 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 28. (k) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:33 and an antisense strand containing the nucleotide sequence of SEQ ID NO:34. In the double-stranded RNA of the present invention, the combination of the sense strand and the antisense strand is more preferably selected from the following double-stranded RNAs: (b) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 7 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 8; (e) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 17 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 18; (f) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 19 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 20; (g) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 25 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 26. (h) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 27 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 28. (k) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:33 and an antisense strand containing the nucleotide sequence of SEQ ID NO:34.

[0027] In yet another embodiment, in the double-stranded RNA of the present invention, a combination of a sense strand and an antisense strand is more preferably selected from the following double-stranded RNAs: (y) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:114 and an antisense strand containing the nucleotide sequence of SEQ ID NO:115. (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (cl) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 316 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 317. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (cw) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:358 and an antisense strand containing the nucleotide sequence of SEQ ID NO:359. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447. In the double-stranded RNA of the present invention, the combination of the sense strand and the antisense strand is more preferably selected from the following double-stranded RNAs: (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (br) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:256 and an antisense strand containing the nucleotide sequence of SEQ ID NO:257. (cd) A dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:288 and an antisense strand containing the nucleotide sequence of SEQ ID NO:289. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353. (dh) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:396 and an antisense strand containing the nucleotide sequence of SEQ ID NO:397. (dq) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:416 and an antisense strand containing the nucleotide sequence of SEQ ID NO:417. (ee) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO:446 and an antisense strand containing the nucleotide sequence of SEQ ID NO:447. In the double-stranded RNA of the present invention, the combination of the sense strand and the antisense strand is more preferably selected from the following: (a1) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 164 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 165. (cu) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 352 and an antisense strand comprising the nucleotide sequence of SEQ ID NO: 353.

[0028] A nucleobase (or nucleobase) of a nucleotide is a heterocyclic pyrimidine or purine compound that is a component of all nucleic acids and includes adenine (A), guanine (G), cytosine (C), thymine (T), and uracil (U). As used herein, "G," "g," "C," "c," "A," "a," "U," "u," and "T" generally refer to nucleobases, nucleosides, nucleotides, and nucleotide mimetics that contain guanine, cytosine, adenine, uracil, and thymine, respectively, as bases.

[0029] As used herein, the term "sequence" or "nucleobase sequence" refers to a string or order of nucleobases and / or nucleotides and / or nucleosides and is described in terms of the standard nomenclature for nucleotides and the notations for modified nucleotides described herein. As used herein, the term "nucleotide" can include modified nucleotides, modified nucleotide mimetics, abasic sites, and surrogate replacement moieties.

[0030] The sense strand and / or the antisense strand may optionally independently comprise an additional 1, 2, 3, 4, 5, or 6 nucleotides (extensions) at the 3' end, or at the 5' end, or at both the 3' and 5' ends of the core sequence. The additional nucleotides of the antisense strand, if present, may or may not be complementary to the corresponding sequence of YTHDC1 mRNA. The additional nucleotides of the sense strand, if present, may or may not be complementary to the corresponding sequence of YTHDC1 mRNA. The additional nucleotides of the antisense strand, if present, may or may not be complementary to the additional nucleotides present in the corresponding sense strand.

[0031] As used herein, an extension comprises 1, 2, 3, 4, 5, or 6 nucleotides located at the 5'-end and / or 3'-end of the core sequence of the sense strand and / or the core sequence of the antisense strand. The extension nucleotides on the sense strand may or may not be complementary to the corresponding nucleotides (core sequence nucleotides or extension nucleotides) of the antisense strand. Conversely, the extension nucleotides on the antisense strand may or may not be complementary to the corresponding nucleotides (core sequence nucleotides or extension nucleotides) of the sense strand. In some embodiments, both the sense and antisense strands of the double-stranded RNA comprise a 3' extension and a 5' extension. In some embodiments, one or more 3' extension nucleotides of one strand are base-paired with one or more 5' extension nucleotides of the other strand. In other embodiments, one or more 3' extension nucleotides of one strand are not base-paired with one or more 5' extension nucleotides of the other strand. In some embodiments, the double stranded RNA has an antisense strand with a 3' extension and a sense strand with a 5' extension.

[0032] In some embodiments, the sense and antisense strands of the double-stranded RNA described herein comprise the same number of nucleotides. In some embodiments, the sense and antisense strands of the double-stranded RNA described herein comprise a different number of nucleotides. In some embodiments, the 5' end of the sense strand and the 3' end of the antisense strand of the double-stranded RNA form a blunt end. The 3' end of the sense strand and the 5' end of the antisense strand of the double-stranded RNA form a blunt end. In some embodiments, both ends of the double-stranded RNA form a blunt end. In some embodiments, neither end of the double-stranded RNA is blunt. In some embodiments, the 5' end of the sense strand and the 3' end of the antisense strand of the double-stranded RNA form an unwound end. In some embodiments, the 3' end of the sense strand and the 5' end of the antisense strand of the double-stranded RNA form an unwound end. In some embodiments, both ends of the double-stranded RNA form an unwound end. In some embodiments, neither end of the double-stranded RNA is an unwound end. As used herein, an uncoiled end refers to an end of a double-stranded RNA molecule where the terminal nucleotides of the two annealed strands are paired (i.e., do not form an overhang) but are not complementary (i.e., form a non-complementary pair). As used herein, an overhang is an extension of one or more unpaired nucleotides located at the end of one strand of a double-stranded RNA molecule. The unpaired nucleotides may be on the sense or antisense strand, resulting in a 3' overhang or a 5' overhang. In some embodiments, the double-stranded RNA molecule comprises a blunt end and an uncoiled end, a blunt end and a 5' overhang end, a blunt end and a 3' overhang end, an uncoiled end and a 5' overhang end, an uncoiled end and a 3' overhang end, two 5' overhang ends, two 3' overhang ends, a 5' overhang end and a 3' overhang end, two uncoiled ends, or two blunt ends.

[0033] As used herein, a "modified nucleotide" is a nucleotide other than a ribonucleotide (2'-hydroxyl nucleotide). Non-limiting examples of modified nucleotides include deoxynucleotides, nucleotide mimics, abasic nucleotides, 2'-modified nucleotides, 3'→3'-linked (inverted) nucleotides, nucleotides containing non-natural bases, bridged nucleotides, nucleotide mimics, and locked nucleotides. Non-limiting examples of 2'-modified nucleotides (i.e., nucleotides having a group other than a hydroxyl group at the 2' position of the five-membered sugar ring) include 2'-O-methyl nucleotides (represented herein by a lowercase "n" in a nucleotide sequence), 2'-deoxy-2'-fluoro nucleotides (represented herein by Nf, but also referred to herein as 2'-fluoro nucleotides), 2'-deoxy nucleotides, and 2'-methoxyethyl (2'-O-2-methoxyethyl) nucleotides. It is not necessary that every position of a given compound be uniformly modified. More than one modification can be incorporated into a single double-stranded RNA, or more than one modification can be incorporated into a single nucleotide of the double-stranded RNA. The sense and antisense strands of the double-stranded RNA can be synthesized and / or modified by methods known in the art. The modification of one nucleotide is independent of the modification of another nucleotide.

[0034] In some embodiments, one or more nucleotides of the double-stranded RNA are linked by a non-standard linkage or backbone (i.e., a modified internucleoside linkage or a modified internucleoside backbone). In some embodiments, the modified internucleoside linkage is a non-phosphate-containing covalent internucleoside linkage. Non-limiting examples of modified internucleoside linkages or modified internucleoside backbones include phosphorothioates, 5'-phosphorothioate groups (represented herein by the lowercase "s" preceding the nucleotide, sN, sn, sNf), and chiral phosphorothioates.

[0035] In some embodiments, the double-stranded RNA comprises one or more modified nucleotides and one or more modified internucleoside linkages. In some embodiments, 2' modified nucleotides are combined with modified internucleoside linkages. For example, in some embodiments, the sense strand of the double-stranded RNA can comprise one, or two, or three, or four phosphorothioate linkages; or the antisense strand of the double-stranded RNA can comprise one, or two, or three, or four phosphorothioate linkages; or both the sense strand and the antisense strand can independently comprise one, or two, or three, or four phosphorothioate linkages.

[0036] The pharmaceutical composition comprising the double-stranded RNA of the present invention may be designed using drug delivery system (DDS) technology for delivery to a target tissue or cell of interest. Methods for designing and producing double-stranded RNA (e.g., siRNA) for delivery to a target tissue or cell of interest are known in the art, including, for example, methods for producing double-stranded RNA with modifications (e.g., chemical modifications) that enable the properties of the double-stranded RNA to be improved in terms of stability, bioavailability, and therapeutic utility (Yuko Ito, STI Horizon, Vol. 5, No. 4, 2019, https: / / doi.org / 10.15108 / stih.00196). In addition, methods for formulating and delivering pharmaceutical compositions comprising double-stranded RNA to living organisms are also well known in the art. For example, but not limited to, the methods described in Signal Transduction and Targeted Therapy, volume 5, Article number: 101, 2020. The pharmaceutical composition comprising the double-stranded RNA of the present invention may or may not comprise, for example, lipid nanoparticles (LNPs) or N-acetylgalactosamine (GalNAc). In certain embodiments, it is preferred to comprise lipid nanoparticles (LNPs) or N-acetylgalactosamine (GalNAc).

[0037] In some embodiments, double-stranded RNA can be expressed from a transcription unit inserted into a vector. The recombinant vector can be a DNA plasmid, a non-viral vector, or a viral vector. The viral vector for expressing double-stranded RNA can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vector capable of expressing double-stranded RNA can be delivered as described above, allowing for transient or stable expression. For example, such a vector can include 1) a transcription initiation region, 2) optionally a transcription termination region, and 3) a nucleic acid sequence encoding at least one strand of double-stranded RNA, which sequence may be linked to the initiation region and the termination region to allow expression and / or delivery of double-stranded RNA. The double-stranded RNA of the present invention may be encoded by the above-mentioned recombinant expression vector, and if encoded by a recombinant expression vector, it is preferably encoded by an adeno-associated virus, retrovirus, adenovirus, or alphavirus vector, and more preferably encoded by an adeno-associated virus vector.

[0038] Adeno-associated viruses are single-stranded DNA viruses consisting of approximately 4,700 bases and belonging to the Parvoviridae family. They are non-enveloped and have a regular icosahedral capsid with a diameter of 20 to 30 nm. They infect cells by recognizing heparan sulfate proteoglycan, a universal component of the cell membrane, and therefore have a wide host range. Any gene or nucleic acid can be inserted into the genome of the adeno-associated virus. Therefore, the gene or nucleic acid of interest can be introduced by infecting a target cell with a genetically modified adeno-associated virus, and the adeno-associated virus can be used as a vector for gene introduction. Furthermore, since the "adeno-associated virus vector" is non-pathogenic and can introduce genes into terminally differentiated, non-dividing cells, it is expected to be applied to gene therapy. It is known that adeno-associated viruses have different infection tropisms for tissues and cells depending on the serotype (Ozawa et al., Drug Delivery System, Vol. 22, No. 6, pp. 643-650, 2007). The substance contained in the pharmaceutical composition containing the double-stranded RNA of the present invention is preferably adeno-associated virus type 5, adeno-associated virus type 7, adeno-associated virus type 8 or adeno-associated virus type 9, and more preferably adeno-associated virus type 8.

[0039] The double-stranded RNA of the present invention can be combined with one or more additional (i.e., second, third, etc.) therapeutic agents. Second therapeutic agents include nucleic acid analog compounds or double-stranded RNA against HBV RNA. Additional therapeutic agents include small molecule compounds, antibodies, vaccines, etc. The double-stranded RNA, with or without one or more additional drugs, can be combined with one or more pharma- ceutically acceptable carriers to form a pharmaceutical composition.

[0040] Pharmaceutically acceptable carriers include excipients, diluents, bulking agents, disintegrants, stabilizers, preservatives, buffers, emulsifiers, fragrances, colorants, sweeteners, thickeners, flavorings, dissolution aids, and other additives. By using one or more of such carriers, the double-stranded RNA can be prepared in the form of tablets, capsules, powders, syrups, granules, pills, suspensions, emulsions, powder preparations, suppositories, eye drops, nasal drops, ear drops, patches, ointments, injections, liquids, lozenges, or elixirs. It is preferable to prepare the double-stranded RNA in the form of a suspension or injection. These double-stranded RNAs can be administered in a number of ways depending on whether local or systemic treatment is desired, for example, intravenously, intra-arterially, subcutaneously, intraperitoneally, subcutaneously (e.g., via an implanted device), or intrasoft tissue. Furthermore, the dosage and frequency of administration of the double-stranded RNA of the present invention can be appropriately selected depending on the patient's sex, body weight, age, severity, symptoms, etc. Usually, for adults, 0.01 to 1000 mg / kg per day may be administered once or in several divided doses by drip infusion or subcutaneous administration.

[0041] The double-stranded RNA of the present invention is useful as a double-stranded RNA for treating hepatitis B in a subject, the double-stranded RNA comprising an inhibitor against the expression of YTHDC1.

[0042] A pharmaceutical composition comprising the double-stranded RNA of the present invention can be used in a method for treating an HBV infection or a disease or disorder caused by HBV infection, comprising administering to a subject (e.g., a human or animal subject) the double-stranded RNA described herein.

[0043] <Uses of double-stranded RNA> The double-stranded RNA of the present invention is useful in the treatment or prevention of HBV infection.

[0044] The double-stranded RNA of the present invention can inhibit the production of HBV proteins, particularly the production of HBs antigen. It can also inhibit HBV gene expression (HBV DNA production). Therefore, the double-stranded RNA of the present invention is useful for treating or preventing HBV infection.

[0045] The present invention relates to the use of the double-stranded RNA of the present invention for inhibiting the production of HBs antigen. The present invention relates to the use of the double-stranded RNA of the present invention for inhibiting HBV DNA production. The present invention relates to the use of the double-stranded RNA of the present invention for inhibiting gene expression of HBV.

[0046] The present invention relates to the use of the double-stranded RNA of the present invention for the treatment or prevention of HBV infection. Disease associated with HBV infection includes progressive liver fibrosis, inflammation and necrosis that progresses to cirrhosis, end-stage liver disease and hepatocellular carcinoma.

[0047] Next, the present invention will be described with reference to test examples, but the present invention is not limited to these. EXAMPLES

[0048] [Test Example 1] (Screening of unmodified dsRNA against YTHDC1) The knockdown effect of the designed unmodified dsRNA on YTHDC1 mRNA and anti-HBV activity were evaluated using HepG2.2.15 cells (Proc Natl Acad Sci USA vol. 84, pp. 1005-1009, 1987), which are cells in which the HBV genome was introduced into the hepatoblastoma cell line HepG2 and which continuously produce the virus, as follows. The following cell culture media were used: DMEM / F-12, GlutaMAX (Invitrogen) + 5 μg / mL insulin (Fujifilm Wako Pure Chemical Industries, Ltd.) + 1% penicillin / streptomycin (Fujifilm Wako Pure Chemical Industries, Ltd.) + 10 mmol / L HEPES (Fujifilm Wako Pure Chemical Industries, Ltd.) + 50 μmol / L hydrocortisone (Sigma) + 10% FBS (Equitech-Bio)

[0049] The dsRNA used for transfection had the sequences shown in Table 1 (designated as duplex numbers #1 to #31). Negative Control No. 1 siRNA (ThermoFisher Scientific) was used as a negative control dsRNA. HepG2.2.15 cells were suspended in the above medium and 2 × 10 5 A HepG2.2.15 cell suspension was prepared at 100 nmol / mL. dsRNA was diluted with Nuclease-Free Water (not DEPC-Treated) (ThermoFisher Scientific) to prepare a 500 nmol / L dsRNA solution. This was further diluted with serum-free medium (ThermoFisher Scientific) to prepare a 100 nmol / L dsRNA solution. 0.3 μL of Lipofectamine RNAiMAX Transfection Reagent (ThermoFisher Scientific) and 4.7 μL of serum-free medium were added to 5 μL of this dsRNA solution, and incubated at room temperature for 5 minutes. 10 μL of this dsRNA-Lipofectamine RNAiMAX Transfection Reagent mixture was diluted with 40 μL of cell culture medium to make 50 μL. This was mixed with 50 μL of the above-mentioned HepG2.2.15 cell suspension, seeded in a 96-well microtiter plate, and incubated at 37°C in a 5% CO2 incubator for 3 days. Next, the medium was replaced with 100 μL / well of the above-mentioned cell culture medium, and incubated at 37°C in a 5% CO2 incubator for another 3 days. The culture supernatant was then collected and cell viability was measured using CellTiter96 AQueous One Solution Cell Proliferation Assay (Promega) (Table 2). After measuring the cell viability, the cells were washed with phosphate-buffered saline (100 μL / well, Fujifilm Wako Pure Chemical), and total RNA was extracted using NucleoSpin 96 RNA (Machrei-Nagel). 7 μL of the extracted total RNA was reverse transcribed using PrimeScript RT Reagent Kit with gDNA Eraser (Takara) to prepare cDNA.YTHDC1 and the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the cDNA were quantified by real-time PCR, and the expression level of YTHDC1 corrected by GAPDH was calculated. The relative value (%) of the expression level of YTHDC1 in each well to which unmodified dsRNA was added to the well to which negative control dsRNA was added was calculated (Table 2). In addition, the amount of HBs antigen in the collected culture supernatant was measured using AlphaLISA Hepatitis B Virus Surface Antigen (HBsAg) Kit (Perkin Elmer). The relative value (%) of each well to which unmodified dsRNA was added to the well to which negative control dsRNA was added was calculated (Table 2).

[0050] [Table 1] JPEG0007644136000002.jpg102170

[0051] [Table 2]

[0052] Among the unmodified dsRNAs against YTHDC1, duplex numbers #1, #4, #6, #7, #9, #10, #13, #14, #15, #16, #17, #18, and #19 reduced YTHDC1 mRNA expression by 30% or more. Among them, duplex numbers #1, #4, #9, #13, #14, #16, and #17 reduced YTHDC1 mRNA expression by 50% or more. Furthermore, while no significant decrease in cell viability (cell viability less than 50%) was observed with these seven types of dsRNA, HBs antigen in the culture supernatant was reduced by about 90% in each case.

[0053] [Test Example 2] (Evaluation of activity of unmodified dsRNA against YTHDC1) Using HepG2.2.15 cells, the activities of dsRNAs with duplex numbers #1, #4, #6, #9, #10, #13, #14, and #17 found in Test Example 1 were evaluated by the following procedure. The following cell culture media were used: DMEM / F-12, GlutaMAX + 5μg / mL insulin + 1% penicillin / streptomycin + 10mmol / L HEPES + 50μmol / L hydrocortisone + 10% FBS The dsRNAs used for transfection had the sequences shown in Table 1 (designated as duplex numbers #1, #4, #6, #9, #10, #13, #14, and #17). In addition, Negative Control No. 1 siRNA was used as a negative control dsRNA. (1) HepG2.2.15 cells were suspended in the above medium and diluted to 2 × 10 5 A HepG2.2.15 cell suspension was prepared at 10 ... Next, the medium was replaced with 100 μL / well of the cell culture medium described above, and the cells were incubated at 37°C in a 5% CO2 incubator for another 3 days. The culture supernatant was then collected, and the cell viability was measured using the CellTiter96 AQueous One Solution Cell Proliferation Assay (Figure 1). The amount of HBs antigen in the collected culture supernatant was measured using the AlphaLISA Hepatitis B Virus Surface Antigen (HBsAg) Kit. The collected culture supernatant was treated with a mixture of Buffer AL (Qiagen) and Proteinase K (ThermoFisher Scientific), and the HBV DNA (HBV DNA) in the resulting solution was quantified by real-time PCR. For each marker, the relative value (%) of each dsRNA-added well to the negative control dsRNA-added well at each concentration was calculated (Figures 2 and 3). Furthermore, the 50% inhibitory concentration (IC50) was calculated for each marker (Tables 3 and 4). (2) HepG2.2.15 cells were suspended in the above medium and diluted to 2 × 10 5 A HepG2.2.15 cell suspension of 100 nmol / mL was prepared. dsRNA was diluted with Nuclease-Free Water (not DEPC-Treated) to prepare a 500 nmol / L dsRNA solution. This was further diluted with serum-free medium to prepare a 100 nmol / L dsRNA solution. 3 μL of Lipofectamine RNAiMAX Transfection Reagent and 47 μL of serum-free medium were added to 50 μL of this dsRNA solution, and incubated at room temperature for 5 minutes. 100 μL of this dsRNA-Lipofectamine RNAiMAX Transfection Reagent mixture was diluted with 400 μL of cell culture medium to make 500 μL. This was mixed with 500 μL of the above-mentioned HepG2.2.15 cell suspension, seeded on a 12-well microtiter plate, and incubated at 37 °C in a 5% CO2 incubator for 3 days. Next, the medium was replaced with 1000 μL / well of the cell culture medium described above, and the cells were incubated at 37 °C in a 5% CO2 incubator for another 3 days. After removing the culture supernatant, the cells were washed with 1000 μL / well of phosphate-buffered saline, and total RNA was extracted using RNeasy Mini Kit (Qiagen). 800 ng of the extracted total RNA was reverse transcribed using PrimeScript RT Reagent Kit with gDNA Eraser to prepare cDNA. YTHDC1 and GAPDH, a housekeeping gene, in the prepared cDNA were quantified by real-time PCR, the YTHDC1 expression level corrected by GAPDH was calculated, and the relative value (%) of the YTHDC1 expression level in each unmodified dsRNA-added well to the YTHDC1 expression level in the negative control dsRNA-added well was calculated (Figure 4).

[0054] A slight decrease in cell viability was observed in duplex number #1 at a concentration of 1.67 nmol / L or more, but no effect on cell viability was observed in #4, #6, #9, #10, #13, #14, or #17 at each concentration (Figure 1). HBs antigen was reduced dose-dependently by each dsRNA of #1, #4, #6, #9, #10, #13, #14, or #17, and the maximum efficacy of each was a reduction in HBs antigen of 90% or more (Figure 2). The IC50 value of the HBs antigen reduction effect of each dsRNA was less than 0.1 nmol / L (Table 3). In addition, HBV DNA was reduced dose-dependently by each dsRNA, and the maximum efficacy of each was a reduction in HBV DNA of 90% or more (Figure 3). The IC50 value of the HBV DNA reduction effect of each dsRNA was less than 0.2 nmol / L (Table 4). Furthermore, duplex numbers #1, #4, #9, #10, #13, #14, and #17 reduced the YTHDC1 expression level to less than 50%, and in particular #4, #9, #10, #13, #14, and #17 reduced the YTHDC1 expression level to less than 70% (Figure 4).

[0055] [Table 3]

[0056] [Table 4]

[0057] [Test Example 3] (Screening of modified dsRNA against YTHDC1) Using HepG2.2.15 cells, the knockdown effect of the designed modified dsRNA on YTHDC1 mRNA and anti-HBV activity were evaluated by the following procedures. The following cell culture media were used: DMEM / F-12, GlutaMAX + 5μg / mL insulin + 1% penicillin / streptomycin + 10mmol / L HEPES + 50μmol / L hydrocortisone + 10% FBS

[0058] The modified dsRNA used for transfection had the sequence shown in Table 5 (designated duplex numbers #32 to #224). In addition, a modified dsRNA against firefly luciferase (sequence shown in Table 5, designated duplex number #225) was used as a negative control dsRNA. HepG2.2.15 cells were suspended in the above medium and 2 × 10 5 A HepG2.2.15 cell suspension of 100 nmol / mL was prepared. dsRNA was diluted with Nuclease-Free Water (not DEPC-Treated) to prepare a 500 nmol / L dsRNA solution. This was further diluted with serum-free medium to prepare a 100 nmol / L dsRNA solution. 0.3 μL of Lipofectamine RNAiMAX Transfection Reagent and 4.7 μL of serum-free medium were added to 5 μL of this dsRNA solution, and incubated at room temperature for 5 minutes. 10 μL of this dsRNA-Lipofectamine RNAiMAX Transfection Reagent mixture was diluted with 40 μL of cell culture medium to make 50 μL. This was mixed with 50 μL of the above-mentioned HepG2.2.15 cell suspension, seeded on a 96-well microtiter plate, and incubated at 37 °C in a 5% CO2 incubator for 3 days. Next, the medium was replaced with 100 μL / well of the above-mentioned cell culture medium, and the cells were incubated at 37°C in a 5% CO2 incubator for another 3 days. The culture supernatant was then collected, and the cell viability was measured using CellTiter96 AQueous One Solution Cell Proliferation Assay (Table 6). After measuring the cell viability, the cells were lysed with ISOGENII (manufactured by Nippon Gene). Total RNA was extracted from the cell lysate using DIRECT-zol-96 MagBead RNA Kit (manufactured by ZYMORESEARCH). 7 μL of the extracted total RNA was reverse transcribed using PrimeScript RT Reagent Kit with gDNA Eraser to prepare cDNA. YTHDC1 and GAPDH in the prepared cDNA were quantified by real-time PCR, the YTHDC1 expression level corrected by GAPDH was calculated, and the relative value (%) of the YTHDC1 expression level in each dsRNA-added well to the YTHDC1 expression level in the negative control dsRNA-added well was calculated (Table 6). In addition, the amount of HBs antigen in the collected culture supernatant was measured using the AlphaLISA Hepatitis B Virus Surface Antigen (HBsAg) Kit.The relative value (%) of each well to which dsRNA was added was calculated relative to the well to which negative control dsRNA was added (Table 6).

[0059] Among the modified dsRNAs against YTHDC1, the duplex numbers are #38, #40, #42, #43, #44, #46, #48, #51, #52, #55, #56, #57, #59, #64, #66, #68, #69, #70, #71, #72, #75, #76, #79, #80, #82, #84, #87, #88, #89, #91, #92, and # 93, #94, #97, #98, #99, #100, #101, #102, #103, #105, #106, #107, #110, #111, #113, #117, #118, #119, #121, #122, #123, #124, #125, #126, #127, #128, #129, #130, #131, #132, #133, #134, #1 35, #140, #141, #142, #143, #144, #145, #146, #147, #148, #151, #154, #156, #158, #161, #165, #166, #168, #172, #173, #174, #175, #176, #178, #179, #180, #182, #184, #186, #189, #190, #191 The expression level of YTHDC1 mRNA was reduced by 30% or more by 123 types of dsRNA: #195, #196, #197, #198, #199, #200, #201, #202, #203, #205, #206, #207, #208, #209, #211, #212, #213, #214, #215, #216, #217, #218, #219, #220, #221, #222, and #223. In addition, the reduction in cell viability (cell viability less than 70%) was not observed by these 123 types of dsRNA. Of these 123 types of dsRNA, the duplex numbers are #38, #42, #43, #44, #46, #57, #59, #66, #69, #72, #79, #82, #84, #87, #89, #91, #92, #93, #97, #99, #100, #101, #105, #106, #107, #110, #111, #117, #121, #122, #124, #125, #126, #127, #128, #129, #130, #131, #132, #140, #141, #142, #143, and #144. The expression level of YTHDC1 mRNA was reduced by more than 50% by 85 types of dsRNA: #145, #147, #148, #151, #154, #158, #172, #174, #175, #176, #178, #179, #180, #182, #186, #189, #190, #195, #197, #198, #199, #200, #202, #203, #205, #206, #207, #208, #209, #212, #213, #214, #215, #216, #217, #218, #219, #220, #221, #222, and #223. Of these 85 types of dsRNA, 36 types of dsRNA, namely, duplex numbers #57, #59, #82, #89, #92, #97, #105, #106, #111, #122, #124, #128, #129, #132, #141, #143, #144, #145, #147, #151, #158, #172, #174, #176, #178, #179, #180, #197, #198, #205, #207, #208, #213, #214, #222, and #223, reduced YTHDC1 mRNA expression by 70% or more. Furthermore, of the 36 types of dsRNA mentioned above, 33 types of dsRNA, namely, duplex numbers #57, #59, #82, #89, #92, #97, #105, #106, #122, #124, #128, #129, #132, #141, #143, #144, #147, #151, #158, #174, #176, #178, #179, #180, #197, #198, #205, #207, #208, #213, #214, #222, and #223, all reduced HBs antigen in the culture supernatant by more than 50%. Among them, 17 types of dsRNAs (duplex numbers: #57, #82, #92, #105, #106, #128, #143, #144, #158, #174, #176, #179, #180, #198, #207, #208, and #223) reduced HBs antigen in the culture supernatant by 70% or more, and 15 types of dsRNAs (duplex numbers: #57, #82, #92, #105, #106, #143, #144, #158, #174, #179, #180, #198, #207, #208, and #223) reduced HBV DNA in the culture supernatant by 50% or more.

[0060] [Table 5] JPEG0007644136000007.jpg252170JPEG0007644136000008.jpg252170JPEG0007644136000009.jpg252170JPEG000 7644136000010.jpg252170JPEG0007644136000011.jpg252170JPEG0007644136000012.jpg252170JPEG00076441360 00013.jpg252170JPEG0007644136000014.jpg252170JPEG0007644136000015.jpg252170JPEG0007644136000016.j pg252170JPEG0007644136000017.jpg252170JPEG0007644136000018.jpg252170JPEG0007644136000019.jpg252170

[0061] [Table 6] JPEG0007644136000021.jpg243170JPEG0007644136000022.jpg243170JPEG0007644136000023.jpg243170 JPEG0007644136000024.jpg243170JPEG0007644136000025.jpg243170JPEG0007644136000026.jpg243170

[0062] [Test Example 4] (Evaluation of activity of modified dsRNA against YTHDC1) Using HepG2.2.15 cells, the activity of 10 types of modified dsRNAs found in Test Example 3, namely, duplex numbers #57, #82, #128, #144, #158, #176, #179, #198, #208, and #223, was evaluated by the following procedure. The following cell culture media were used: DMEM / F-12, GlutaMAX + 5μg / mL insulin + 1% penicillin / streptomycin + 10mmol / L HEPES + 50μmol / L hydrocortisone + 10% FBS The dsRNA used for transfection had the sequence shown in Table 5 (designated duplex numbers #57, #82, #128, #144, #158, #176, #179, #198, #208, and #223). In addition, a modified dsRNA against firefly luciferase (sequence shown in Table 5, designated duplex number #225) was used as a negative control dsRNA. HepG2.2.15 cells were suspended in the above medium and 2 × 10 5 A HepG2.2.15 cell suspension was prepared at 1000 cells / mL. dsRNA was diluted with Nuclease-Free Water (not DEPC-Treated) to prepare dsRNA solutions at 0.686 to 1500 nmol / L, 3-fold dilution series. These were further diluted with serum-free medium to prepare dsRNA solutions at 0.137 to 300 nmol / L, 3-fold dilution series. 0.3 μL of Lipofectamine RNAiMAX Transfection Reagent and 4.7 μL of serum-free medium were added to 5 μL of dsRNA solution at each concentration, and incubated at room temperature for 5 minutes. 10 μL of this dsRNA-Lipofectamine RNAiMAX Transfection Reagent mixture was diluted with 40 μL of cell culture medium to make 50 μL. This was mixed with 50 μL of the above-mentioned HepG2.2.15 cell suspension, seeded on a 96-well microtiter plate, and incubated at 37 °C in a 5% CO2 incubator for 3 days. Next, the medium was replaced with 100 μL / well of the above-mentioned cell culture medium, and the cells were further incubated at 37°C in a 5% CO2 incubator for 3 days. The culture supernatant was then collected, and the cell viability was measured using the CellTiter96 AQueous One Solution Cell Proliferation Assay (Figure 5). The amount of HBs antigen in the collected culture supernatant was measured using the AlphaLISA Hepatitis B Virus Surface Antigen (HBsAg) Kit. The relative value (%) of each dsRNA-added well to the negative control dsRNA-added well was calculated (Figure 6). Furthermore, the collected culture supernatant was treated with a mixture of Buffer AL and Proteinase K, and the HBV DNA (HBV DNA) in the obtained solution was quantified by real-time PCR. The relative value (%) of each dsRNA-added well to the negative control dsRNA-added well was calculated (Figure 7). After measuring the cell viability, the cells were lysed with ISOGENII. Total RNA was extracted from cell lysates using the DIRECT-zol-96 MagBead RNA Kit.7 μL of the extracted total RNA was reverse transcribed using PrimeScript RT Reagent Kit with gDNA Eraser to prepare cDNA. YTHDC1 and GAPDH in the prepared cDNA were quantified by real-time PCR, and the YTHDC1 expression level corrected by GAPDH was calculated. The relative value (%) of the YTHDC1 expression level in each dsRNA-added well to the YTHDC1 expression level in the negative control dsRNA-added well was calculated (Figure 8).

[0063] The ten dsRNAs, duplex numbers #57, #82, #128, #144, #158, #176, #179, #198, #208, and #223, did not affect cell viability at any concentration (Figure 5). Each dsRNA reduced HBs antigen in a roughly dose-dependent manner, with the maximum efficacy of all reducing HBs antigen by 60% or more (Figure 6). Among them, duplex numbers #82, #128, #144, #176, #198, #208, and #223 all showed a maximum efficacy of reducing HBs antigen by 70% or more, and duplex numbers #82 and #176 in particular showed a maximum efficacy of reducing HBs antigen by 80% or more. The IC50 value of the HBs antigen reduction effect of each dsRNA was less than 5 nmol / L (Table 7). Furthermore, each dsRNA reduced HBV DNA in a roughly dose-dependent manner, with each showing a maximum efficacy of reducing HBV DNA by 50% or more (Figure 7). The IC50 values ​​for the HBV DNA reduction effect of each dsRNA were all less than 5 nmol / L (Table 8). Furthermore, each dsRNA reduced YTHDC1 expression levels in a roughly dose-dependent manner, with each showing a maximum efficacy of reducing YTHDC1 expression levels by 70% or less (Figure 8). The IC50 values ​​for the YTHDC1 expression reduction effect of each dsRNA were all less than 5 nmol / L (Table 9).

[0064] [Table 7]

[0065] [Table 8]

[0066] [Table 9] [Industrial Applicability]

[0067] The double-stranded RNA of the present invention exhibits excellent anti-HBV activity and is useful as an anti-hepatitis B virus agent.

Claims

1. An inhibitor of hepatitis B virus protein production, comprising a double-stranded RNA which inhibits the production of a hepatitis B virus protein, the double-stranded RNA being formed from a sense strand and an antisense strand, the combination of the sense strand and the antisense strand forming the double-stranded RNA being selected from the following: (a) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 1 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 2; (b) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 7 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 8; (c) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 11 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 12; (d) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 13 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 14; (e) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 17 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 18; (f) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 19 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 20; (g) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 25 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 26; (h) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 27 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 28; (j) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 31 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 32; (k) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 33 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 34; (l) a dsRNA formed from a sense strand containing the nucleotide sequence of SEQ ID NO: 35 and an antisense strand containing the nucleotide sequence of SEQ ID NO: 36; (m) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO:37 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

38. (Y) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 114 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

115. (ah) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 150 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

151. (al) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 164 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

165. (cr) A dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO:346 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

347. (cw) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 358 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

359. (cy) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 364 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

365. (dh) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO:396 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

397. (dw) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 430 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

431. (dy) a dsRNA formed from a sense strand comprising the nucleotide sequence of SEQ ID NO: 434 and an antisense strand comprising the nucleotide sequence of SEQ ID NO:

435.

2. The inhibitor of the production of a Hepatitis B virus protein according to claim 1, which comprises a modified nucleotide.

3. The inhibitor of production of a Hepatitis B virus protein according to any one of claims 1 to 2, wherein the HBV protein is HBs antigen (HBsAg) protein.

4. The inhibitor of hepatitis B virus protein production according to any one of claims 1 to 3, which further reduces the expression level of HBV DNA.

5. The inhibitor of hepatitis B virus protein production according to any one of claims 1 to 4, which further reduces the expression amount of completely closed double-stranded DNA (cccDNA).

6. The inhibitor of hepatitis B virus protein production according to any one of claims 1 to 5, wherein the double-stranded RNA is an RNA interference agent.

7. A pharmaceutical composition for inhibiting the production of Hepatitis B virus proteins, comprising an inhibitor of the production of Hepatitis B virus proteins according to any one of claims 1 to 6.

8. The pharmaceutical composition of claim 7 , wherein the double-stranded RNA is encoded by a recombinant expression vector, and the recombinant expression vector is an adeno-associated virus vector.

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