RNA inhibitor for inhibiting expression of insulin-like growth factor 1 receptor

Abstract

The present application relates to an RNA inhibitor that inhibits the expression of an insulin-like growth factor 1 receptor (IGF-1R) gene. The RNA inhibitor contains an antisense strand that forms a complementary region with at least 15 consecutive nucleotides in the mRNA encoding the IGF-1R (SEQ ID NO: 828), wherein the complementary region contains 0, 1, 2, 3, 4 or 5 mismatches.

Description

[Corrected 20.02.2025 in accordance with Rule 26] RNA inhibitors for inhibiting the expression of insulin-like growth factor 1 receptor Technical Field

[0001] The present application relates to the field of biomedicine, and specifically to an RNA inhibitor and a conjugate thereof for inhibiting the expression of the insulin-like growth factor 1 receptor gene. Background Art

[0002] RNA interference (RNAi) technology refers to a highly evolutionarily conserved phenomenon in which double-stranded RNA (dsRNA) induces the efficient and specific degradation of homologous mRNAs. RNA molecules inhibit the expression of certain genes by destroying specific mRNAs. Because RNAi can specifically knock out or shut down the expression of specific genes, it has rapidly become one of the most sought-after research tools in gene function research and gene therapy. It has been widely used to explore gene function and treat metabolic diseases, infectious diseases, and malignant tumors.

[0003] Thyroid eye disease (TED) is an autoimmune disease caused by hyperthyroidism. It is caused by autoantigens activating the IGF-1R-mediated signaling complex on cells within the orbit, leading to the body's own immune system attacking the tissues around and behind the eyes. The disease is characterized by enlargement of the extraocular muscles, adipose tissue, and connective tissue, and can produce symptoms such as dry eyes, bulging eyes, retraction of the eyelids, double vision, strabismus, stinging pain with a gritty sensation, easy tearing, redness and swelling, and difficulty with eye movement. Severe cases can lead to disfigurement and impaired vision.

[0004] IGF-1R (Insulin-like growth factor 1 receptor) is a tetrameric transmembrane receptor tyrosine kinase found on the surface of human cells. It serves as the cell surface receptor for IGF-1 (the hormone insulin-like growth factor 1). Studies have shown that IGF-1R is widely expressed in normal human tissues and is highly expressed in thyroid eye disease (TED). IGF-1R activation stimulates cell proliferation, survival, transformation, metastasis, and angiogenesis, while IGF-1R inhibition can alleviate and improve TED symptoms. Tepezza (Teprotumumab), the world's first drug for the treatment of chronic (inactive) thyroid eye disease (TED) developed by Horizon Therapeutics, has been approved for marketing in the United States by the US Food and Drug Administration. This drug is a fully human monoclonal antibody (mAb) and a targeted inhibitor of IGF-1R. Unlike other biologics for the treatment of TAO, such as anti-CD20 antibodies, which only reduce inflammation, Tepezza effectively reduces eye bulging in addition to eliminating inflammation. Patients treated with Tepezza experienced an unprecedented reduction in proptosis, which previously could only be treated surgically after active disease resolved. However, Tepezza also had significant safety concerns, with the most common adverse reactions including muscle spasms, hearing loss, and hyperglycemia. These adverse reactions may be due to the systemic administration of the drug.

[0005] The development of locally administered IGF-1R siRNA offers a solution to the current safety concerns of monoclonal antibodies. Due to their long-lasting efficacy, siRNA-based drugs can allow for longer dosing cycles, leading to improved compliance. Furthermore, IGF-1R is a key driver of tumor growth and progression, and the development of siRNA targeting IGF-1R has broad potential applications in cancer treatment. Summary of the Invention

[0006] The present application provides an RNA inhibitor that inhibits the expression of the IGF-1R gene, providing an effective new treatment for thyroid eye disease, osteoarthritis, and neuropathic pain. IGF-1R is a tetrameric transmembrane receptor tyrosine kinase found on the surface of human cells and is the cell surface receptor for IGF-1. The RNA inhibitor of IGF-1R gene expression provided by the present application can destroy the function of IGF-1R mRNA as a translation template, inhibit the expression of IGF-1R protein, and alleviate and improve the symptoms of thyroid eye disease, osteoarthritis, and neuropathic pain. The siRNA drug provided by the present application has one or more of the following therapeutic advantages: strong efficacy, small side effects, long duration of efficacy, low dosage, small injection reaction, and good expected patient compliance. On the other hand, siRNA drugs can improve drug accessibility and improve drug delivery efficiency through reasonable formulations.

[0007] On the one hand, the present application provides an RNA inhibitor for inhibiting the expression of the insulin-like growth factor 1 receptor (IGF-1R) gene, comprising an antisense strand, wherein the antisense strand forms a complementary region with at least 15 consecutive nucleotides in the mRNA encoding IGF-1R (SEQ ID NO: 828), and the complementary region has 0, 1, 2, 3, 4 or 5 mismatches. Preferably, the complementary region is 15-30 nucleotide pairs in length, and more preferably 17-23 nucleotide pairs in length.

[0008] In some embodiments, the antisense strand forms a complementary region with 15, 16, 17, 18, 19, 20, 21, 22 or 23 consecutive nucleotides starting from the 5' end of any one of the following positions in the mRNA encoding IGF-1R (SEQ ID NO: 828):

[0009] Nucleotide position 1118, nucleotide position 1120, nucleotide position 1154, nucleotide position 1367, nucleotide position 1407, nucleotide position 1415, nucleotide position 1532, nucleotide position 1625, nucleotide position 1627, nucleotide position 1628, nucleotide position 1631, nucleotide position 3356, nucleotide position 3357, nucleotide position 3359, nucleotide position 3792, nucleotide position 4198, nucleotide position 4200, nucleotide position 4208, nucleotide position 4685, nucleotide position 5246, nucleotide position 5392, nucleotide position 5393, nucleotide position 6329, nucleotide position 6332, nucleotide position 6333 , nucleotide No. 6334, nucleotide No. 10215, nucleotide No. 10738, nucleotide No. 10740, nucleotide No. 10770, nucleotide No. 10772, nucleotide No. 10773, nucleotide No. 10946, nucleotide No. 10956, nucleotide No. 10957, nucleotide No. 1418, nucleotide No. 1541, nucleotide No. 2050, nucleotide No. 2055, nucleotide No. 2056, nucleotide No. 2233, nucleotide No. 2234, nucleotide No. 2456, nucleotide No. 2591, nucleotide No. 2603, nucleotide No. 2606, nucleotide No. 2609, nucleotide No. 2776, nucleotide No. 2890, Nucleotide No. 2906, Nucleotide No. 2909, Nucleotide No. 2957, Nucleotide No. 2960, Nucleotide No. 3008, Nucleotide No. 3195, Nucleotide No. 3398, Nucleotide No. 3608, Nucleotide No. 3609, Nucleotide No. 3650, Nucleotide No. 3660, Nucleotide No. 3815, Nucleotide No. 3882, Nucleotide No. 3996, Nucleotide No. 4067, Nucleotide No. 4121, Nucleotide No. 4122, Nucleotide No. 4292, Nucleotide No. 4293, Nucleotide No. 4295, Nucleotide No. 4516, Nucleotide No. 4519, Nucleotide No. 5163, Nucleotide No. 5309, Nucleotide No. 5310, Nucleotide No. 5377, Nucleotide No. 5380, Nucleotide No. 5381, Nucleotide No. 5663, Nucleotide No. 5700, Nucleotide No. 5711, Nucleotide No. 5712, Nucleotide No. 5721, Nucleotide No. 5896, Nucleotide No. 5898, Nucleotide No. 6203, Nucleotide No. 6236, Nucleotide No. 6237, Nucleotide No. 6240, Nucleotide No. 6245, Nucleotide No. 6288, Nucleotide No. 6299, Nucleotide No. 6305, Nucleotide No. 6309, Nucleotide No. 6310, Nucleotide No. 6343, Nucleotide No. 6363, Nucleotide No. 6365, Nucleotide No. 6366, Nucleotide No. 6373,Nucleotide No. 6379, Nucleotide No. 6381, Nucleotide No. 6431, Nucleotide No. 6434, Nucleotide No. 6496, Nucleotide No. 6518, Nucleotide No. 6755, Nucleotide No. 6760, Nucleotide No. 6883, Nucleotide No. 6885, Nucleotide No. 6947, Nucleotide No. 6948, Nucleotide No. 6949, Nucleotide No. 6952, Nucleotide No. 6953, Nucleotide No. 6954, Nucleotide No. 7574, Nucleotide No. 7621, Nucleotide No. 7764, Nucleotide No. 7830, Nucleotide No. 7906, Nucleotide No. 8540, Nucleotide No. 8836, Nucleotide No. 8892, Nucleoside No. 8975 Acid, nucleotide No. 8977, nucleotide No. 9163, nucleotide No. 9175, nucleotide No. 9186, nucleotide No. 9265, nucleotide No. 9267, nucleotide No. 9340, nucleotide No. 9347, nucleotide No. 9352, nucleotide No. 9356, nucleotide No. 9478, nucleotide No. 9480, nucleotide No. 9606, nucleotide No. 9611, nucleotide No. 9634, nucleotide No. 9680, nucleotide No. 9681, nucleotide No. 9724, nucleotide No. 9725, nucleotide No. 9814, nucleotide No. 10015, nucleotide No. 10016, nucleotide No. 10189, nucleotide No. 10244, nucleotide No. 10 nucleotide 10526, nucleotide 10528, nucleotide 10534, nucleotide 10535, nucleotide 10537, nucleotide 10540, nucleotide 10541, nucleotide 10679, nucleotide 10694, nucleotide 10717, nucleotide 10728, nucleotide 10729, nucleotide 10730, nucleotide 10754, nucleotide 10763, nucleotide 10766, nucleotide 10942, nucleotide 11034, nucleotide 11078, nucleotide 11126, nucleotide 11156 , nucleotide 11159, nucleotide 11160, nucleotide 11292, nucleotide 11340, nucleotide 11344, nucleotide 11383, nucleotide 11395, nucleotide 11789, nucleotide 11864, nucleotide 11865, nucleotide 11939, nucleotide 12092, nucleotide 12116, nucleotide 12130, nucleotide 12154, nucleotide 12156, nucleotide 12177, nucleotide 12195, nucleotide 1164, nucleotide 1163, nucleotide 1162, nucleotide 1161, nucleotide 1160,Nucleotide 1165, nucleotide 1166, nucleotide 1167, nucleotide 1168.

[0010] In some embodiments, the antisense strand and the mRNA encoding IGF-1R (SEQ ID NO: 828) form a complementary region between nucleotides 1160 and 1188, between nucleotides 6235 and 6465, or between nucleotides 6329 and 6455, counting from the 5' end.

[0011] In some embodiments, the RNA inhibitor is ribonucleic acid, and further, the RNA inhibitor is single-stranded ribonucleic acid or double-stranded ribonucleic acid.

[0012] In some embodiments, the RNA inhibitor is an antisense oligonucleotide (ASO), shRNA, miRNA, or siRNA.

[0013] In some embodiments, it comprises a sense chain capable of forming a complementary double strand with the antisense chain, wherein the antisense chain comprises a sequence that forms a duplex complementary region with at least 15 consecutive nucleotides in the sense chain sequence, and the duplex complementary region has 0, 1, 2, 3, 4 or 5 mismatches. Preferably, the duplex complementary region is 15-30 nucleotide pairs in length, more preferably 17-23 nucleotide pairs.

[0014] In some embodiments, wherein the sense strand and the antisense strand are present on two different nucleic acid strands, preferably the RNA inhibitor is siRNA or shRNA, more preferably the RNA inhibitor is siRNA.

[0015] In some embodiments, wherein the sense strand and the antisense strand are present on the same nucleic acid strand, preferably the RNA inhibitor is shRNA.

[0016] In some embodiments, the total length of the sense strand is 15-50 nucleotides, preferably the total length of the sense strand is 16-30 nucleotides, and more preferably the total length of the sense strand is 17, 18, 19, 20 or 21 nucleotides.

[0017] In some embodiments, the total length of the antisense strand is 19-50 nucleotides, preferably the total length of the antisense strand is 19-30 nucleotides, and more preferably the total length of the antisense strand is 21, 22, 23, 24, 25, 26 or 27 nucleotides.

[0018] In some embodiments, the sense strand and antisense strand each independently optionally comprise a 3' or 5' overhang of 1, 2, or 3 nucleotides.

[0019] In some embodiments, both the sense and antisense strands have 3' overhangs of 1-3 nucleotides in length, or the sense strand has a 3' or 5' overhang of 1-3 nucleotides in length, or the antisense strand has a 3' or 5' overhang of 1-3 nucleotides in length.

[0020] In some embodiments, the antisense strand of the RNA comprises at least 15 consecutive nucleotides of the sequence of any one of SEQ ID NOs: 199-400.

[0021] In some embodiments, the RNA inhibitor, wherein the sense strand comprises at least 15 consecutive nucleotides in the sequence of any one of SEQ ID NOs: 1-198.

[0022] In some embodiments, the RNA inhibitor comprises a duplex selected from the group consisting of: ds-n1, ds-n2, ds-n3, ds-n4, ds-n5, ds-n6, ds-n7, ds-n8, ds-n9, ds-n10, ds-n11, ds-n12, ds-n13, ds-n14, ds-n15, ds-n16, ds-n17, ds-n18, ds-n19, ds-n20, ds-n21, ds-n22, ds-n23, ds-n24, ds-n25, ds-n26, ds-n27, ds-n28, ds-n29, ds-n30, ds-n31, ds-n32, ds-n33, ds-n34, s-n33, ds-n34, ds-n35, ds-n36, ds-n37, ds-n38, ds-n39, ds-n40, ds-n41, ds-n42, ds-n43, ds-n44, ds-n45, ds-n46, ds-n47, ds-n48, ds-n49, ds-n50 , ds-n51, ds-n52, ds-n53, ds-n54, ds-n55, ds-n56, ds-n57, ds-n58, ds-n5 9. ds-n60, ds-n61, ds-n62, ds-n63, ds-n64, ds-n65, ds-n66, ds-n67, ds-n6 8. ds-n69, ds-n70, ds-n71, ds-n72, ds-n73, ds-n74, ds-n75, ds-n76, ds-n 77. ds-n78, ds-n79, ds-n80, ds-n81, ds-n82, ds-n83, ds-n84, ds-n85, ds- n86, ds-n87, ds-n88, ds-n89, ds-n90, ds-n91, ds-n92, ds-n93, ds-n94, ds -n95, ds-n96, ds-n97, ds-n98, ds-n99, ds-n100, ds-n101, ds-n102, ds-n10 3. ds-n104, ds-n105, ds-n106, ds-n107, ds-n108, ds-n109, ds-n110, ds-n 111, ds-n112, ds-n113, ds-n114, ds-n115, ds-n116, ds-n117, ds-n118, ds- n119, ds-n120, ds-n121, ds-n122, ds-n123, ds-n124, ds-n125, ds-n126, d s-n127, ds-n128, ds-n129, ds-n130, ds-n131, ds-n132, ds-n133, ds-n134,ds-n135, ds-n136, ds-n137, ds-n138, ds-n139, ds-n140, ds-n141, ds-n142, ds-n 143, ds-n144, ds-n145, ds-n146, ds-n147, ds-n148, ds-n149, ds-n150, ds-n151, ds-n152, ds-n153, ds-n154, ds-n155, ds-n156, ds-n157, ds-n158, ds-n159, ds-n 160, ds-n161, ds-n162, ds-n163, ds-n164, ds-n165, ds-n166, ds-n167, ds-n168, ds-n169, ds-n170, ds-n171, ds-n172, ds-n173, ds-n174, ds-n175, ds-n176, ds-n 177, ds-n178, ds-n179, ds-n180, ds-n181, ds-n182, ds-n183, ds-n184, ds-n185, ds-n186, ds-n187, ds-n188, ds-n189, ds-n190, ds-n191, ds-n192, ds-n193, ds-n 194, ds-n195, ds-n196, ds-n197, ds-n198, ds-n201, ds-n202, ds-n203, ds-n204. ,

[0023] In some embodiments, an RNA inhibitor comprises a duplex selected from the group consisting of: ds-n17, ds-n23, ds-n31, ds-n39, ds-n45, ds-n58, ds-n60, ds-n64, ds-n67, ds-n68, ds-n73, ds-n74, ds-n76, ds-n77, ds-n80, ds-n81, ds-n83, ds-n84, ds-n86, ds-n88, ds-n89, ds-n98 , ds-n99, ds-n103, ds-n104, ds-n112, ds-n113, ds-n115, ds-n153, ds-n154, ds-n156, ds-n157, ds-n163, ds-n 164, ds-n165, ds-n188, ds-n190, ds-n191, ds-n192, ds-n193, ds-n194, ds-n195, ds-n196, ds-n197, ds-n198.

[0024] In some embodiments, the NA inhibitor comprises a duplex selected from the group consisting of ds-n17, ds-n23, ds-n31, ds-n39, ds-n45, ds-n58, ds-n60, ds-n64, ds-n67, ds-n68, ds-n73, ds-n74, ds-n76, ds-n77, ds-n80, ds-n81, ds-n83 , ds-n84, ds-n86, ds-n88, ds-n89, ds-n98, ds-n99, ds-n103, ds-n104, ds-n112, ds-n11 3. ds-n115, ds-n153, ds-n154, ds-n156, ds-n157, ds-n163, ds-n164, ds-n165, ds-n188.

[0025] In some embodiments, the RNA inhibitor comprises a duplex selected from the group consisting of: ds-n23, ds-n58, ds-n67, ds-n86, ds-n89, ds-n98, ds-n99, ds-n103, ds-n190, ds-n198, and ds-n113.

[0026] In some embodiments, the RNA inhibitor is characterized in that the antisense strand further includes region B1 at the 3' end, and the sense strand further includes region A1 at the 5' end, wherein region A1 is 0-6 nucleotides and region B1 is 0-6 nucleotides.

[0027] In some embodiments, the RNA inhibitor is characterized in that the region B1 is 0, 1, 2, 3, 4, 5 or 6 nucleotides; preferably, the region B1 is 0 or 2 nucleotides.

[0028] In some embodiments, the RNA inhibitor is characterized in that the region A1 is 0, 1, 2, 3, 4, 5 or 6 nucleotides; preferably, the region A1 is 0 or 2 nucleotides.

[0029] In some embodiments, the RNA inhibitor is characterized in that the antisense strand further includes region X2 at the 5' end, and the sense strand or sense nucleic acid further includes region X1 at the 3' end, and region X1 and region X2 are complementary.

[0030] In some embodiments, the RNA inhibitor, wherein X1 is A, U, modified A or modified U, and X2 is U, A, modified U or modified A.

[0031] In some embodiments, the RNA inhibitor is characterized in that the antisense strand comprises X2, Y, Z and N in sequence from 3' to 5' direction at the 5' end, wherein X2 and Y are independently A, U, modified A or modified U, Z is G or modified G, and N contains at least one nucleotide.

[0032] In some embodiments, in the RNA inhibitor, X2, Y, and Z are AAG, AUG, UUG, or UAG, or partially or completely modified sequences thereof, in sequence from 3' to 5'.

[0033] In some embodiments, in the RNA inhibitor, X2, Y, and Z are AAG, AUG, UUG, or UAG, or partially or completely modified sequences thereof, in sequence from 3' to 5'.

[0034] In some embodiments, the RNA inhibitor, wherein N is C or modified C.

[0035] In some embodiments, in the RNA inhibitor, X2, Y, Z and N are AAGC and UAGC in sequence from 3' to 5', or the above sequences that have been partially or completely modified.

[0036] In some embodiments, the RNA inhibitor, wherein the antisense strand comprises at least 15 consecutive nucleotides in the sequence of any one of SEQ ID NOs: 401-413.

[0037] In some embodiments, the RNA inhibitor is selected from ds-n190-1, ds-n191-1, ds-n192-1, ds-n193-1, ds-n194-1, ds-n195-1, ds-n196-1, ds-n197-1, ds-n198-1, ds-n201-1, ds-n202-1, ds-n203-1, and ds-n204-1.

[0038] In some embodiments, the RNA inhibitor, wherein at least one nucleotide in the RNA inhibitor is a modified nucleotide.

[0039] In some embodiments, the RNA inhibitor, wherein at least 70%, 80%, 90%, 95% of the nucleotides in the RNA inhibitor are modified nucleotides; preferably all nucleotides are modified nucleotides.

[0040] In some embodiments, the RNA inhibitor, wherein the modification comprises a combination of one or more of the following: 2'-OMe (2'-O-methyl) modification, 2'-F (2'-deoxy-2'-fluoro) modification, 2'-O-MOE (2'-O-methoxyethyl) modification, 2'-deoxy (2'-d) modification, 5'-morpholine (5'-Mo) modification, unlocked nucleic acid (UNA) modification, glycol nucleic acid (GNA) modification, locked nucleic acid (LNA) modification, tricyclic DNA (tcDNA) modification, (S)-constrained ethyl bicyclic nucleic acid ((S)-cEt-BNA) modification, phosphorothioate (PS) modification, phosphorodithioate (PS2) modification, methylphosphonate (MP) modification, methoxypropylmethylphosphonate (MOP) modification, peptide nucleic acid (PNA) modification, 5'-(E)-vinyl phosphate (VP) modification (V P), N6-methyladenosine (m6A) modification, 5-methylcytidine (m5C) modification, 3-methyluridine (m3U) modification, 5-methyluridine (m5U) modification, pseudouridine modification, 2-thiouridine (s2U) modification, propyne uridine (5-pU) modification, inverted abasic nucleotide (invAB) modification by bonding the 5' or 3' end of the nucleotide, replacing the nucleotide with an inverted abasic nucleotide (invAb) modification, replacing the nucleotide with a 2,4-difluoromethylphenyl ribonucleotide (rF) modification or replacing the nucleotide with a (S)-glycerol nucleic acid modification, preferably a 2'-OMe modification, a 2'-F modification, a 2'-deoxy modification, a VP modification, a 5'-MP modification, a PS modification, a PS2 modification, an MP modification, a MOP modification, an M06 modification, an invAb modification or an invAB modification.

[0041] In some embodiments, the RNA inhibitor comprises a phosphate or a phosphate mimetic at the 3' or 5' end of the antisense strand, or a phosphate or a phosphate mimetic at the 3' or 5' end of the sense strand.

[0042] In some embodiments, the RNA inhibitor described above, wherein the phosphate mimetic comprises 5'-(E)-vinylphosphonate, 5'-methylphosphonate, (S)-5'-C-methyl analogs, and 5'-phosphorothioate (5'-PS).

[0043] In some embodiments, the RNA inhibitor, wherein the 5'-end of the antisense strand comprises (M06) modification:

[0044] In some embodiments, the RNA inhibitor has an invAB modification at the 3' or 5' end of the antisense strand, or an invAB modification at the 3' or 5' end of the sense strand.

[0045] In some embodiments, the RNA inhibitor, wherein the inverted abasic nucleotide or M06 is linked to the 3' or 5' end of the antisense strand or the 3' or 5' end of the sense strand via phosphorothioate.

[0046] In some embodiments, the RNA inhibitor has the following modification combination

[0047] a) the sense strand comprises a first sequence of 16-21 nt in length, wherein the first sequence comprises the following modifications: 2'-F modifications are present at nucleotides 9, 10, and 11, counting from the 5' end;

[0048] b) the antisense strand comprises a second sequence of 16-21 nt in length, wherein the second sequence includes the following modifications: counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, a phosphorothioate linkage between nucleotides 2 and 3, and a phosphorothioate linkage between nucleotides 3 and 4; and counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2; and counting from the 5' end, there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16.

[0049] In some embodiments, the RNA inhibitor has the following modification combination

[0050] a) the sense strand comprises a first sequence of 16-21 nt in length, wherein the first sequence comprises the following modifications: counting from the 5' end, nucleotides at positions 9, 10, and 11 have 2'-F modifications, and the remaining nucleotides have 2'-Ome modifications;

[0051] b) the antisense strand comprises a second sequence of 16-21 nt in length, wherein the second sequence includes the following modifications: counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, a phosphorothioate linkage between the 2nd and 3rd nucleotides, and a phosphorothioate linkage between the 3rd and 4th nucleotides; and counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides; and counting from the 5' end, there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16, and the rest are 2'-Ome modifications.

[0052] In some embodiments, the RNA inhibitor has the following modification combination

[0053] a) the sense strand comprises a first sequence of 16-21 nt in length, wherein the first sequence comprises the following modifications: 2'-F modifications are present at nucleotides 9, 10, and 11, counting from the 5' end;

[0054] b) the antisense strand comprises a second sequence of 16-21 nt in length, wherein the second sequence includes the following modifications: counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, a phosphorothioate linkage between nucleotides 2 and 3, and a phosphorothioate linkage between nucleotides 3 and 4; and counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2; and counting from the 5' end, there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16.

[0055] In some embodiments, the RNA inhibitor has a first sequence length of 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, or 21 nt, preferably 21 nt.

[0056] In some embodiments, the length of the second sequence of the RNA inhibitor is preferably 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, preferably 21 nt.

[0057] In some embodiments, the RNA inhibitor has the following modification combination

[0058] a) The sense strand is 21 nt long and includes the following modifications: 2'-F modifications are present at nucleotides 9, 10, and 11, counting from the 5' end;

[0059] b) The antisense strand is 21 nt in length and includes the following modifications: counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, a phosphorothioate linkage between nucleotides 2 and 3, and a phosphorothioate linkage between nucleotides 3 and 4; and, counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2; and, counting from the 5' end, there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16.

[0060] In some embodiments, the RNA inhibitor has the following modification combination

[0061] a) The positive strand is 21 nt long and includes the following modifications: Counting from the 5' end, nucleotides 9, 10, and 11 have 2'-F modifications, and the rest have 2'-Ome modifications;

[0062] b) The antisense strand is 21 nt in length and includes the following modifications: counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, a phosphorothioate linkage between nucleotides 2 and 3, and a phosphorothioate linkage between nucleotides 3 and 4; and, counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2; and, counting from the 5' end, there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16, and the remaining nucleotides have a 2'-Ome modification.

[0063] In some embodiments, the RNA inhibitor, wherein the antisense strand comprises at least 15 consecutive nucleotides in the sequence of any one of SEQ ID NOs: 624-812.

[0064] In some embodiments, the RNA inhibitor, wherein the sense strand comprises at least 15 consecutive nucleotides of the sequence of any one of SEQ ID NOs: 414-602.

[0065] In some embodiments, the RNA inhibitor is selected from:

[0066] ds-m1、ds-m2、ds-m3、ds-m4、ds-m5、ds-m6、ds-m7、ds-m8、ds-m9、ds-m10、ds-m11、ds-m12、ds-m13、ds-m14、ds-m15、ds-m16、ds-m17、ds-m18、ds-m19、ds-m20、ds-m21、ds-m22、ds-m23、ds-m24、ds-m25、ds-m26、ds-m27、ds-m28、ds-m29、ds-m30、ds-m31、ds-m32、ds-m33、ds-m34、ds-m35、ds-m36、ds-m37、ds-m38、ds-m39、ds-m40、ds-m41、ds-m42、ds-m43、ds-m44、ds-m45、ds-m46、ds-m47、ds-m48、ds-m49、ds-m50、ds-m51、ds-m52、ds-m53、ds-m54、ds-m55、ds-m56、ds-m57、ds-m58、ds-m59、ds-m60、ds-m61、ds-m62、ds-m63、ds-m64、ds-m65、ds-m66、ds-m67、ds-m68、ds-m69、ds-m70、ds-m71、ds-m72、ds-m73、ds-m74、ds-m75、ds-m76、ds-m77、ds-m78、ds-m79、ds-m80、ds-m81、ds-m82、ds-m83、ds-m84、ds-m85、ds-m86、ds-m87、ds-m88、ds-m89、ds-m90、ds-m91、ds-m92、ds-m93、ds-m94、ds-m95、ds-m96、ds-m97、ds-m98、ds-m99、ds-m100、ds-m101、ds-m102、ds-m103、ds-m104、ds-m105、ds-m106、ds-m107、ds-m108、ds-m109、ds-m110、ds-m111、ds-m112、ds-m113、ds-m114、ds-m115、ds-m116、ds-m117、ds-m118、ds-m119、ds-m120、ds-m121、ds-m122、ds-m123、ds-m124、ds-m125、ds-m126、ds-m127、ds-m128、ds-m129、ds-m130、ds-m131、ds-m132、ds-m133、ds-m134、ds-m135、ds-m136、ds-m137、ds-m138、ds-m139, ds-m140, ds-m141, ds-m142, ds-m143, ds-m144, ds-m145, ds-m146, ds-m147, ds-m148, ds-m149, ds-m150, ds-m15 1. ds-m152, ds-m153, ds-m154, ds-m155, ds-m156, ds-m157, ds-m158, ds-m159, ds-m160, ds-m161, ds-m162, ds-m163, ds-m 164, ds-m165, ds-m166, ds-m167, ds-m168, ds-m169, ds-m170, ds-m171, ds-m172, ds-m173, ds-m174, ds-m175, ds-m176, ds -m177, ds-m178, ds-m179, ds-m180, ds-m181, ds-m182, ds-m183, ds-m184, ds-m185, ds-m186, ds-m187, ds-m188, ds-m189. ,

[0067] In some embodiments, the RNA inhibitor, wherein

[0068] a) the sense strand comprises a third sequence of 18-21 nt in length, the third sequence comprising the following modifications: counting from the 5' end, a phosphorothioate linkage exists between nucleotides 1 and 2 of the sense strand, a phosphorothioate linkage exists between nucleotides 2 and 3, nucleotides 9, 10, 11, and 18 of the sense strand have 2'-F modifications, and the remaining nucleotides have 2'-OMe modifications; counting from the 3' end, a phosphorothioate linkage exists between nucleotides 1 and 2, and a phosphorothioate linkage exists between nucleotides 2 and 3;

[0069] b) the antisense strand comprises a fourth sequence of 19-26 nt in length, wherein the fourth sequence includes the following modifications: counting from the 5' end, nucleotides 1, 2, 5, 9, 17, and 19 of the antisense strand are 2'-F modified, the rest are 2'-OMe modified, and there is a phosphorothioate linkage between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides.

[0070] In some embodiments, the RNA inhibitor, wherein the length of the third sequence is preferably 18nt, 19nt, 20nt, 21nt, 22nt, 21nt, preferably 21nt.

[0071] In some embodiments, the length of the fourth sequence of the RNA inhibitor is preferably 19 nt, 20 nt, 21 nt, 22 nt, 23 nt, 24 nt, 25 nt, 26 nt, preferably 26 nt.

[0072] In some embodiments, any one of the RNA inhibitors, wherein

[0073] a) The sense strand is 21 nt long. Counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, and between nucleotides 2 and 3. Nucleotides 9, 10, 11, and 18 of the sense strand are 2'-F modified, and the rest are 2'-OMe modified. Counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2, and between nucleotides 2 and 3.

[0074] b) The antisense strand is 26 nt in length. Counting from the 5' end, nucleotides 1, 2, 5, 9, 17, and 19 of the antisense strand have 2'-F modifications, and the rest have 2'-OMe modifications. There is a phosphorothioate linkage between the 4th and 5th nucleotides. Counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2, and between nucleotides 2 and 3.

[0075] In some embodiments, the RNA inhibitor, wherein the antisense strand comprises at least 15 consecutive nucleotides in the sequence of any one of SEQ ID NOs: 813-827.

[0076] In some embodiments, the RNA inhibitor, wherein the sense strand comprises at least 15 consecutive nucleotides in the sequence of any one of SEQ ID NOs: 603-617.

[0077] In some embodiments, the RNA inhibitor comprises a duplex selected from the group consisting of:

[0078] ds-m190, ds-m191, ds-m192, ds-m193, ds-m194, ds-m195, ds-m196, ds-m 197, ds-m198, ds-m199, ds-m200, ds-m201, ds-m202, ds-m203 and ds-m204.

[0079] In some embodiments, the RNA inhibitor further comprises a delivery system, wherein the delivery system is conjugated to the sense chain and / or antisense chain, and the delivery system enables the RNA inhibitor to reach the target RNA in the target tissue to produce a gene silencing effect.

[0080] In some embodiments, the target tissue is ocular tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

[0081] In some embodiments, the eye tissue is the optic nerve, trabecular meshwork, proximal canal tissue, ganglion, episcleral vein, Schlemm's canal or peripheral eye tissue. Preferably, the joint tissue comprises cartilage tissue, joint connective tissue, and bone tissue; preferably, the central nervous tissue comprises spinal cord tissue and brain tissue; preferably, the peripheral nervous tissue comprises intra-articular nerve tissue and muscle nerve tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

[0082] In some embodiments, the ocular tissue is retinal ganglion, endothelial cells, periocular muscle, or periocular fat.

[0083] In some embodiments, the delivery systems are each independently conjugated to one or more internal locations of the double-stranded RNA.

[0084] In some embodiments, the internal position is on a nucleobase, a sugar ring, a methylphosphonate bond, a phosphorothioate diester bond, or a phosphodiester bond.

[0085] In some embodiments, the delivery system is a lipophilic structure, which includes a lipophilic group and a linker, and the lipophilic group is connected to the double-stranded ribonucleic acid through the linker.

[0086] In some embodiments, the lipophilic structure is selected from aliphatic, alicyclic and polyalicyclic compounds.

[0087] In some embodiments, the lipophilic structure contains saturated or unsaturated C16 or C22.

[0088] In some embodiments, the linker is selected from a single bond, an ether, a thioether, a urea, a carbonate, an amine, an amide, a maleimide-thioether, a disulfide, a phosphodiester, a sulfonamide bond, a product of a click reaction, and a carbamate.

[0089] In some embodiments, the delivery system is

[0090] In some embodiments, the delivery system is conjugated to the 5' end and / or 3' end of the antisense strand or antisense nucleic acid fragment.

[0091] In some embodiments, the delivery system is conjugated to the 5' end and / or 3' end of the sense strand or sense nucleic acid fragment.

[0092] In some embodiments, wherein the delivery system is conjugated to the 5' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 3' end of the sense strand or sense nucleic acid fragment, the two ligands are the same or different.

[0093] In some embodiments, wherein the delivery system is conjugated to the 3' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 5' end of the sense strand or sense nucleic acid fragment, the two ligands are the same or different.

[0094] In some embodiments, wherein the delivery system is conjugated to the 5' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 5' end of the sense strand or sense nucleic acid fragment, the two ligands are the same or different.

[0095] In some embodiments, wherein the delivery system is conjugated to the 3' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 3' end of the sense strand or sense nucleic acid fragment, the two ligands are the same or different.

[0096] In some embodiments, the number of delivery systems is 1, 2, 3, 4, 5, or 6.

[0097] In some embodiments, the antisense strand comprises at least 15 consecutive nucleotides of a sequence as set forth in any one of SEQ ID NOs: 822-827.

[0098] In some embodiments, the sense strand comprises at least 15 consecutive nucleotides of the sequence of any one of SEQ ID NOs: 618-623.

[0099] In some embodiments, the RNA inhibitor is selected from: Z1, Z2, Z3, Z4, Z5, and Z6.

[0100] On the other hand, the present application provides a pharmaceutical composition comprising the RNA inhibitor, and / or a physiologically acceptable excipient and / or carrier and / or diluent.

[0101] In some embodiments, it is characterized in that the pharmaceutically acceptable carrier includes or is selected from aqueous carriers, liposomes, high molecular polymers or polypeptides.

[0102] On the other hand, the present application provides an RNA inhibitor targeting IGF-1R and its pharmaceutical composition for use in the preparation of a drug for treating IGF-1R-related diseases or pathologies; preferably, the RNA inhibitor is siRNA; more preferably, the siRNA is administered to a local or lesion area tissue of a subject in need.

[0103] In some embodiments, the IGF-1R-related disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels.

[0104] In some embodiments, the IGF-1R-related disease or pathology comprises thyroid eye disease, osteoarthritis, and neuropathic pain.

[0105] In some embodiments, the RNA inhibitor and pharmaceutical composition thereof are the RNA inhibitor described in any one of embodiments 1-73 or the pharmaceutical composition described in any one of embodiments 74-75.

[0106] On the other hand, the present application provides a use of an RNA inhibitor or the pharmaceutical composition in the preparation of a medicament for preventing or treating a disease or pathology or reducing the risk of a disease or symptom.

[0107] In another aspect, the present application provides a method for preventing or treating IGF-1R-related diseases or symptoms, comprising administering an effective amount of an RNA inhibitor and a pharmaceutical composition thereof to a subject in need thereof.

[0108] In some embodiments, the RNA inhibitor and the pharmaceutical composition thereof are the RNA inhibitor or the pharmaceutical composition.

[0109] In some embodiments, the RNA inhibitor, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is administered to the subject by intraorbital injection, intraarticular injection, intrathecal injection, subcutaneous administration, intravenous administration, oral administration, rectal administration, or intraperitoneal administration.

[0110] In some embodiments, the method comprises administering to a local or focal area of ​​tissue in a subject in need thereof.

[0111] In some embodiments, the local or lesion area tissue of the subject in need thereof includes eye tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

[0112] In some embodiments, the eye tissue is the optic nerve, trabecular meshwork, proximal canal tissue, ganglion, episcleral vein, Schlemm's canal or peripheral eye tissue. Preferably, the joint tissue comprises cartilage tissue, joint connective tissue, and bone tissue; preferably, the central nervous tissue comprises spinal cord tissue and brain tissue; preferably, the peripheral nervous tissue comprises intra-articular nerve tissue and muscle nerve tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

[0113] In some embodiments, the eye tissue is retinal ganglion, endothelial cells, peripheral ocular muscle or peripheral ocular fat.

[0114] In some embodiments, the IGF-1R-related disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels.

[0115] In some embodiments, the IGF-1R-related disease or pathology comprises thyroid eye disease, osteoarthritis, and neuropathic pain.

[0116] Those skilled in the art will readily appreciate other aspects and advantages of the present application from the detailed description below. The detailed description below merely illustrates and describes exemplary embodiments of the present application. As will be appreciated by those skilled in the art, the content of this application enables those skilled in the art to modify the disclosed specific embodiments without departing from the spirit and scope of the invention to which this application relates. BRIEF DESCRIPTION OF THE DRAWINGS

[0117] The specific features of the invention involved in this application are shown in the appended claims. The features and advantages of the invention involved in this application can be better understood by referring to the exemplary embodiments described in detail below and the accompanying drawings. A brief description of the drawings is as follows:

[0118] FIG1 shows the PD results of intraorbital administration of the RNA inhibitor described in the present application to cynomolgus monkeys.

[0119] FIG2 shows the structure of the 3' end or 5' end of the sense strand or antisense strand of the RNA inhibitor described in the present application. DETAILED DESCRIPTION

[0120] The following describes the implementation of the present invention through specific embodiments. People familiar with this technology can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0121] Definition of terms

[0122] In this application, "optional", "optional" or "optionally" are equivalent in meaning and mean that the event or situation described thereafter may or may not occur, and the description includes both cases where the event or situation occurs and cases where it does not occur. For example, "optionally substituted alkyl" or "alkyl is optionally substituted" includes "alkyl" (H on the alkyl is not replaced / replaced by a non-H substituent) and "substituted alkyl" (H on the alkyl is replaced / replaced by a non-H substituent). As used herein, it will be understood by those skilled in the art that for any group comprising one or more substituents, these groups are not intended to introduce any substitution or substitution pattern that is sterically impractical, synthetically unfeasible and / or inherently unstable. For example, "optionally modified" includes both unmodified and modified, and further, "nucleotides are optionally modified" includes both unmodified nucleotides and modified nucleotides.

[0123] In this application, when any variable (e.g., a substituent R, such as a nucleic acid that is modified) occurs more than once in the composition or structure of a compound, its definition in each case is independent. For example, if a group is substituted with 0-2 Rs, the group can optionally be substituted with up to two Rs, and each R has independent options. For another example, when multiple nucleotides are modified, each nucleotide is independently optionally modified, and the type and amount of modification of each nucleotide can be the same or different.

[0124] In this application, the terms "RNA inhibitor," "iRNA," "siRNA," "RNAi agent," "iRNA agent," and "RNA interference agent" are used interchangeably and generally refer to an agent comprising RNA, as that term is defined herein, which can mediate targeted cleavage of RNA transcripts by forming an RNA-induced silencing complex (RISC) within a cell. The RNA inhibitor directs sequence-specific degradation of mRNA via a process known as RNA interference (RNAi). The RNA inhibitors described herein can regulate or inhibit gene expression of the IGF-1R gene in a cell, and in some embodiments, the cell can be a cell of a mammalian subject. In some embodiments, the RNA inhibitor can regulate or inhibit the mRNA sequence of the IGF-1R gene (NM_000875.5 Homo sapiens insulin-like growth factor 1 receptor), comprising the sequence set forth in SEQ ID NO: 828.

[0125] In certain embodiments, the "RNA inhibitor" used in this application is a single-stranded siRNA (ssRNAi), which can be introduced into a cell or organism to inhibit the target mRNA. The single-stranded RNA inhibitor can bind to the RISC endonuclease Argonaute 2, which then cuts the target mRNA. Single-stranded RNA inhibitors are generally 15 to 30 nucleotides and are chemically modified. The design and testing of single-stranded siRNAs are described in U.S. Patent No. 8,101,348 and Lima et al. (2012) Cell 150: 883-894, the entire contents of each of which are incorporated herein by reference. Any antisense nucleotide sequence described in this application can be used as a single-stranded siRNA described in this application or chemically modified by the methods described in Lima et al. (2012) Cell 150: 883-894.

[0126] In certain embodiments, the "RNA inhibitor" used in this application is shRNA. The term "shRNA" (i.e., short hairpin RNA) used herein refers to an artificial single-stranded interfering RNA molecule that contains the sense and / or antisense strands of an "siRNA duplex" in a stem-loop or hairpin structure. The stem of this hairpin structure is typically in the range of 19 to 29 nucleotides, and the loop is typically in the range of 4 to 15 nucleotides (see, e.g., Siolas, D. et al. (2004) Nat. Biotechnol. 23, 227-231). Typically, shRNA molecules are encoded in a DNA gene expression vector under the control of an RNA polymerase III promoter (e.g., U6 promoter).

[0127] In certain embodiments, the "RNA inhibitor" used in this application is miRNA. The term "miRNA" or "microRNA" is used herein according to its common sense in the art and refers to a small, non-protein coding RNA molecule that is expressed in a variety of eukaryotic organisms, including mammals, and participates in RNA-based gene regulation. A mature, fully processed miRNA is about 15 to about 30 nucleotides in length. A representative group of known endogenous miRNA species is described in the publicly available miRBase sequence database, described in Griffith-Jones et al., Nucleic Acids Research, 2004, 32: D109-D111 and Griffith-Jones et al., Nucleic Acids Research, 2006, 34: D 140-D144, and accessible on the World Wide Web of the Wellcome Trust Sanger Institute website. The fully processed mature miRNA publicly available on the miRBase sequence database is incorporated herein by reference. A representative group of miRNA is also included in Table 1 below. Each mature miRNA is partially complementary to one or more messenger RNA (mRNA) molecules that are targets of the miRNA, thereby regulating the expression of target-associated genes.

[0128] In certain embodiments, the " RNA inhibitor " used in the present application is an ASO. The term "ASO" (i.e., antisense nucleic acid) used herein refers to a nucleic acid molecule having a sense strand and / or antisense strand of an " siRNA duplex " that interacts with a target RNA through RNA-RNA or RNA-DNA or RNA-PNA (protein nucleic acid; Egholm et al., 1993Nature 365,566), and changes the activity of the target RNA through steric interaction or through target recognition mediated by RNase H (for review, see Stein and Cheng, 1993Science 261,1004 and Woolf et al., U.S. Patent number 5,849,902). Generally, antisense molecules are complementary to the target sequence along a single adjacent sequence of the antisense molecule. However, in certain embodiments, the antisense molecule can be bound to a substrate so that the substrate molecule forms a ring, and / or the antisense molecule can be bound so that the antisense molecule forms a ring. Thus, an antisense molecule can be complementary to two (or even more) non-contiguous substrate sequences, or two (or even more) non-contiguous sequence portions of an antisense molecule can be complementary to a target sequence or both. For a review of current antisense strategies, see Schmajuk et al., 1999, J. Biol. Chem., 274, 21783-21789, Delihas et al., 1997, Nature, 15, 751-753, Stein et al., 1997, Antisense NADrug Dev., 7, 151, Crooke, 2000, Methods Enzymol., 313, 3-45; Crooke, 1998, Biotech. Genet. Eng. Rev., 15, 121-157, Crooke, 1997, Ad. Pharmacol., 40, 1-49. In addition, antisense DNA or antisense modified by 2'-MOE and other modifications as known in the art can be used for targeting RNA by DNA-RNA interaction, thereby activating RNase H, which digests the target RNA in the duplex. Antisense oligonucleotides can include one or more RNase H activation regions that can activate the RNase H cleavage of the target RNA. Antisense DNA can be chemically synthesized or genetically expressed using single-stranded DNA gene expression vectors or their equivalents. Antisense molecules of the present invention can be chemically modified as generally known in the art or as described herein.

[0129] In certain embodiments, the "RNA inhibitor" used herein is double-stranded RNA and is referred to herein as a "siRNA duplex," "double-stranded RNA inhibitor," "double-stranded siRNA molecule," "double-stranded siRNA," or "dsRNA." The term "siRNA duplex" refers to a complex of ribonucleic acid molecules having a duplex structure comprising two antiparallel and partially complementary nucleic acid strands, each strand oriented in a "sense" and "antisense" orientation relative to the target RNA (i.e., the IGF-1R gene). In some embodiments, the siRNA duplex directs sequence-specific degradation of an mRNA (e.g., the mRNA sequence of the IGF-1R gene) via a process known as RNA interference (RNAi).

[0130] In general, most of the nucleotides in each chain of an RNA inhibitor are ribonucleotides. In the absence of an explanation, they are ribonucleotides. However, as described in detail herein, each or both of the two chains may also include one or more non-ribonucleotides, such as a deoxyribonucleotide and / or a modified nucleotide. In addition, "RNA inhibitors" may include chemically modified ribonucleotides. These modifications may include all types of modifications disclosed herein or known in the art. Any such modification as used in an RNA inhibitor molecule is encompassed by "RNA inhibitors" for the purposes of this specification and the claims.

[0131] The duplex structure can be any length that allows for specific degradation of the desired target RNA by the RISC pathway, and can be in the range of about 19 to 36 base pairs in length, for example, about 19-30 base pairs in length, for example, about 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 base pairs in length. Ranges and lengths intermediate to the above ranges and lengths are also included as part of this application. In certain embodiments, the RNA inhibitor of the present application is a dsRNA having 15-23 nucleotides in each strand that interacts with a target RNA sequence (e.g., an IGF-1R gene) to guide the cleavage of the target RNA. In certain embodiments, the RNA inhibitor of the present application is a dsRNA of 24-30 nucleotides that interacts with a target RNA sequence (e.g., an mRNA sequence of an IGF-1R gene) to guide the cleavage of the target RNA.

[0132] In this application, the term "antisense strand" generally refers to the strand of an RNA inhibitor (e.g., a duplex siRNA) that is substantially complementary to a target nucleic acid (e.g., an mRNA sequence of a target genomic sequence, including pre-mRNA and mRNA molecules, for example, the mRNA sequence of the IGF-1R gene). The term "region of complementarity" as used in this application generally refers to the region of the antisense strand that is substantially complementary to the mRNA sequence of the IGF-1R gene used in this application. When the region of complementarity is not completely complementary, mismatches can occur within the interior or terminal regions of the molecule. Generally, the most tolerated mismatches are in the terminal regions, for example, within 5, 4, 3, or 2 nucleotides of the 5' end and / or 3' end.

[0133] In the present application, the term "mismatch" refers to situations such as pairing of non-adenine (A) with thymine (T), pairing of non-adenine (A) with uracil (U), pairing of non-guanine (G) with cytosine (C), no hydrogen bond formation between bases of relative nucleotides, and lack of bases between relative nucleotides.

[0134] In this application, the term "antisense nucleic acid fragment" refers to a continuous fragment on the antisense strand, and the fragment can be 15-35 nucleotides in length. For example, the antisense nucleic acid fragment can be a fragment of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, or 34 nucleotides in length on the antisense strand. The antisense nucleic acid fragment is complementary to a continuous nucleotide fragment of any length in the mRNA encoding IGF-1R. In some embodiments, at least three consecutive nucleotide fragments in the antisense nucleic acid fragment are complementary to a continuous nucleotide fragment of a corresponding length in the mRNA encoding IGF-1R.

[0135] In this application, the term "sense strand" generally refers to a strand of an RNA inhibitor that includes a region that is substantially complementary to the region of the antisense strand as defined herein. The "sense" strand is sometimes referred to as a "sense" strand, a "passenger" strand, or an "anti-guide" strand. By means of their sequence, the antisense strand targets the desired mRNA, while the sense strand targets different targets. Therefore, if the antisense strand is incorporated into RISC, the correct target is targeted. The incorporation of the sense strand can result in off-target effects. These off-target effects can be limited by using modifications or using 5' end caps on the sense strand.

[0136] In this application, the term "sense nucleic acid fragment" refers to a continuous fragment on the sense strand, and the fragment length can include 15-35 nucleotides. For example, the sense nucleic acid fragment can be a fragment of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 or 34 nucleotides in length on the sense strand.

[0137] In this application, the term "complementary" refers to the ability of a polynucleotide comprising an RNA inhibitor antisense strand or antisense nucleic acid fragment to hybridize (form base pair hydrogen bonds) and form a duplex or double helix structure with a polynucleotide comprising an RNA inhibitor sense strand or sense nucleic acid fragment or IGF-1R mRNA under certain conditions. Complementary sequences include Watson-Crick base pairs or non-Watson-Crick base pairs, and include natural or modified nucleotides or nucleotide mimetics, as long as the above requirements regarding their hybridization ability are met. "Complementary" does not necessarily require that the bases on each nucleoside have complementarity, and some mismatches may occur in some cases.

[0138] As used herein, the term "fully complementary" generally means that all (100%) of the bases in the contiguous sequence of the antisense strand or antisense nucleic acid fragment of an RNA inhibitor will hybridize to the same number of bases in the contiguous sequence of the sense strand or sense nucleic acid fragment of an RNA inhibitor or IGF-1R mRNA. The contiguous sequence may comprise all or a portion of the above sequence. As used herein, "partially complementary" generally means that in a hybridizing nucleobase sequence pair, at least about 70% of the bases in the contiguous sequence of the antisense strand or sense nucleic acid fragment of an RNA inhibitor will hybridize to the same number of bases in the contiguous sequence of the sense strand or sense nucleic acid fragment of an RNA inhibitor or IGF-1R mRNA. As used herein, the terms "complementary," "fully complementary," and "substantially complementary" can be used with respect to base matching between the sense strand or sense nucleic acid fragment and the antisense strand or antisense nucleic acid fragment of an RNA inhibitor, or between the antisense strand or antisense nucleic acid fragment of an RNA inhibitor and the sequence of IGF-1R mRNA. Sequence identity or complementarity is independent of modification. For example, a and Af are complementary to U (or T) and identical to A for the purposes of determining identity or complementarity.

[0139] As used herein, the term "nucleotide" refers to a 5-carbon sugar (ribose or deoxyribose), a phosphate group, and a base (natural or non-natural base), and is intended to include unmodified (i.e., natural) nucleotides and modified nucleotides. In certain embodiments, the nucleotide is an unmodified ribonucleotide. In certain embodiments, the ribonucleotide is a 3'-ribonucleotide. In certain embodiments, the ribonucleotide is a 5'-ribonucleotide. In certain embodiments, the modified or unmodified nucleotide may optionally be further modified.

[0140] Natural nucleotides are composed of natural bases, natural ribose, and phosphate. As used herein, natural nucleotides refer to adenine ribonucleotides, adenine deoxyribonucleotides, guanine ribonucleotides, guanine deoxyribonucleotides, cytosine ribonucleotides, cytosine deoxyribonucleotides, uracil ribonucleotides, thymine ribonucleotides, or thymine deoxyribonucleotides. The term "ribonucleotide" refers to a nucleotide having a hydroxyl group at the 2' position of the sugar portion of the nucleotide. "Deoxyribonucleoside" refers to a nucleotide having a hydrogen at the 2' position of the sugar portion of the nucleotide.

[0141] The natural bases of RNA include A (adenine), G (guanine), C (cytosine), U (uracil) and T (thymine). As used herein, the mark "d" before a monomer (such as the nucleotides A, U, C, G and T, etc.) indicates that the monomer is 2'-deoxy modified. As used herein, the mark "f" after a monomer (such as the nucleotides A, U, C, G and T, etc.) indicates that the monomer is 2'-deoxy-2'-fluoro modified (2'-F modified). As used herein, the mark "GNA-" before a monomer (such as the nucleotides A, U, C, G and T, etc.) indicates that the monomer is ethylene glycol nucleic acid modified (GNA modified). As used herein, Tgn is the code for GNA-T, which is equivalent. As used herein, lowercase letters (a, u, c, g, t, etc.) indicate that the nucleotide represented by its corresponding uppercase letter (A, U, C, G and T, etc.) is modified by 2'-O-methyl (2'-OMe).

[0142] In the present invention, unless otherwise specified, "G", "C", "A", "T" and "U" refer to guanine ribonucleotide, cytosine ribonucleotide, adenine ribonucleotide, thymine ribonucleotide and uracil ribonucleotide, respectively, and the structure is as follows:

[0143] In the present invention, the mark "d" before a nucleotide (A, U, C, G, and T, etc.) indicates that the nucleotide has been 2'-deoxy modified. For example, the structure of a 2'-deoxy modified nucleotide is as follows:

[0144] In the present invention, the mark "f" after the nucleotide (A, U, C, G and T, etc.) indicates that the nucleotide is modified with 2'-deoxy-2'-fluoro (2'-F modification). For example, the structure of the 2'-F modified nucleotide is as follows:

[0145] In the present invention, the mark "GNA-" before a nucleotide (A, U, C, G, and T, etc.) indicates that the nucleotide is modified with ethylene glycol nucleic acid (GNA modification). In the present invention, Tgn is the code for GNA-T, which is equivalent. Exemplary structures of nucleotides modified with GNA are as follows:

[0146] In the present invention, lowercase letters (a, u, c, g, t, etc.) indicate that the nucleotides represented by their corresponding uppercase letters (A, U, C, G, and T, etc.) are modified with 2'-O-methyl (2'-OMe). For example, the structure of a 2'-OMe-modified nucleotide is as follows:

[0147] In the present invention, the invAB modification refers to the bonding of an inverted abasic nucleotide to the 5' end or the 3' end of a nucleotide.

[0148] For example, The structure after modification by invAB:

[0149] In the present invention, invAb modification refers to the modification of replacing a nucleotide with an inverted abasic nucleotide (invAb). Structure after modification by invAb:

[0150] In the present invention, VP modification refers to modification of the 5' position of a nucleotide with 5'-(E)-vinyl phosphate. For example, the structures of U, u, and dU after modification are as follows:

[0151] In the present invention, the mark "*" between monomers (such as nucleotides A, U, C, G and T, etc.) indicates that the two monomers are connected by a phosphorothioate bond (i.e., a phosphorothioate diester bond), i.e., modified by phosphorothioate (PS).

[0152] In the present invention, the absence of an "*" mark between monomers (such as nucleotides A, U, C, G and T, etc.) indicates that the two nucleotides are connected by a phosphate bond (i.e., a phosphodiester bond).

[0153] For example, "5'-AdUgCf*dT-3'" means that starting from the 5' end of the sequence, position 1 is an adenine ribonucleotide, position 2 is a uracil deoxyribonucleotide, position 3 is a 2'-methoxy-modified guanine ribonucleotide, position 4 is a 2'-fluorine-modified cytosine ribonucleotide, and position 5 is a thymine deoxyribonucleotide connected to position 4 through a phosphorothioate bond.

[0154] As used herein, "optional", "optionally" or "optionally" are equivalent and mean that the event or situation described thereafter may or may not occur, and that the description includes instances where the event or situation occurs and instances where it does not occur. For example, "optionally substituted alkyl" or "alkyl is optionally substituted" includes "alkyl" (H on the alkyl is not substituted / replaced by a non-H substituent) and "substituted alkyl" (H on the alkyl is substituted / replaced by a non-H substituent). As used herein, it will be understood by those skilled in the art that for any group comprising one or more substituents, these groups are not intended to introduce any substitution or substitution pattern that is sterically impractical, synthetically unfeasible and / or inherently unstable. For example, "optionally modified" includes both unmodified and modified, and further, "nucleotides are optionally modified" includes both unmodified nucleotides and modified nucleotides.

[0155] As used herein, when any variable (e.g., a substituent R, e.g., a nucleic acid is modified) occurs more than once in a composition or structure of a compound, its definition at each occurrence is independent. For example, if a group is substituted with 0-2 Rs, then the group may be optionally substituted with up to two Rs, and each occurrence of R has independent options. For another example, when multiple nucleotides are modified, each nucleotide is independently optionally modified, and the type and number of modifications to each nucleotide may be the same or different.

[0156] As used herein, unless otherwise indicated, “comprises,” “includes,” “is at least,” “has,” “has,” “has at least,” “contains,” or the like are open-ended expressions and mean that in addition to the listed elements, components, or steps, other unspecified elements, components, or steps may also be included.

[0157] As used herein, "complementary" or "reverse complementary" are used interchangeably and refer to a structural relationship between two nucleotides (e.g., on two opposing nucleic acid chains or on opposing regions of a single nucleic acid chain) that allows the two nucleotides to form base pairs with each other (e.g., the purine nucleotides of a nucleic acid that are complementary to the pyrimidine nucleotides of the opposing nucleic acid can form base pairs together by forming hydrogen bonds with each other). In some embodiments of the present application, complementary nucleotides can form base pairs in a Watson-Crick manner or in any other manner that allows the formation of a stable duplex. In some embodiments of the present application, two nucleic acid chains may have multiple double-stranded regions that form complementarity. In some embodiments of the present application, in DNA, adenine (A) always pairs with thymine (T), and in RNA, adenine (A) pairs with uracil (U); and guanine (G) always pairs with cytosine (C). In some embodiments of the present application, complementary nucleotides may also include or be completely formed from non-Watson-Crick base pairs and / or base pairs formed from non-natural and modified nucleotides, such non-Watson-Crick base pairs include but are not limited to G:U wobble base pairing or Hoogstein base pairing. In some embodiments of the present application, a nucleotide comprising hypoxanthine as its base may be base paired with a nucleotide comprising adenine, cytosine or uracil. In some embodiments of the present application, a nucleotide containing uracil, guanine or adenine may be replaced in the nucleotide sequence of the present application by a nucleotide containing, for example, inosine (in the application, the capital letter "I" may represent hypoxanthine base, inosine or a nucleotide containing inosine according to its contextual meaning) (this replacement is referred to as I modification). In some embodiments of the present application, adenine and cytosine anywhere in the oligonucleotide can be replaced by guanine and uracil, respectively, to form GU wobble base pairing with the target mRNA.

[0158] The degree of complementarity of one oligonucleotide to another is called complementarity, which is measured by the percentage of bases in each chain that can form hydrogen bonds with each other, which is determined by established base pairing rules. An oligonucleotide sequence does not need to be "completely complementary" (i.e., "perfectly complementary") to its corresponding nucleic acid sequence. In some embodiments, a first nucleotide sequence is considered to be complementary to a second nucleotide sequence if it exhibits at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence complementarity with the second nucleotide sequence. In an exemplary embodiment, 18 of the 20 core bases of the first nucleotide sequence are paired with the corresponding regions of the second nucleotide sequence, achieving 90% complementarity. Non-complementary core bases, also referred to as "mismatches," may be clustered or scattered between complementary bases and do not need to be adjacent to each other or adjacent to complementary core bases.

[0159] As used herein, "mismatch" includes, but is not limited to:

[0160] 1) Two opposing (independent natural or unnatural) nucleotides (other than AT, AU, or GC) pair;

[0161] 2) No hydrogen bonding occurs between two opposing (independent natural or non-natural) nucleotides;

[0162] 3) The absence of a base between two opposing (independent natural or unnatural) nucleotides.

[0163] In some embodiments, the mismatch comprises wobble base pairing and Hoogstein base pairing.

[0164] The term "fully complementary" refers to a first nucleotide sequence and a second nucleotide sequence that form a hybrid consisting only of Watson-Crick base pairs in the fully complementary region. An oligonucleotide that is "fully complementary" may include an internal region (e.g., at least 7, 8, 9, or 10 nucleotides) that is fully complementary to the target RNA.

[0165] As used herein, a blocking group refers to a group that can be conjugated to an oligonucleotide provided herein, for example, at the 5' end of the antisense strand, which can reduce or inhibit exonuclease metabolism. In certain embodiments, the blocking group can reduce or inhibit the RNA interference effect of the oligonucleotide. In certain embodiments, the blocking group is cleaved from the oligonucleotide before providing the RNA interference effect. Examples of blocking groups include, but are not limited to, abasic nucleotide residues, reverse abasic nucleotide residues, MO3, and MO6.

[0166] As used herein, double-stranded nucleotide reagents can be optionally conjugated with one or more blocking groups. Blocking groups can be connected to sense strand, antisense strand or two chains at 3' end, 5' end or both ends. In certain embodiments, blocking groups are conjugated to antisense strand, specifically to 5' end of antisense strand. In certain embodiments, blocking groups are conjugated to oligonucleotide (e.g., 5' end of antisense strand) by nucleotide bonds, and internucleotide bonds are optionally modified as described herein. In certain embodiments, blocking groups are connected to double-stranded nucleotide reagents by phosphorothioate. In certain embodiments, blocking groups are connected to double-stranded nucleotide reagents by phosphodiester bonds.

[0167] In the present invention, M06 modification refers to the bonding of monomers at the 5' or 3' end of a monomer (such as a nucleotide). For example, The structure modified by M06: In the present invention, (M06) It is an M06 monomer In some embodiments, (M06) is represented by Bonded to a nucleotide.

[0168] In this application, the terms "nucleic acid" and "polynucleotide" are used interchangeably and refer to a polymeric form of nucleotides (deoxyribonucleotides or ribonucleotides or their analogs) of any length. The polynucleotide can have any three-dimensional structure and can perform any function. The polynucleotide can contain any one or more modifications or substitutions described in this application or known in the art at one or more bases, sugars and / or phosphates. In some embodiments, the modified nucleotides can be methylated nucleotides or nucleotide analogs. In some embodiments, the nucleotide structure can be modified before or after the assembly of the polynucleotide polymer. The polynucleotide can be modified after polymerization, for example by coupling with a labeling component. The polynucleotide polymer can be blocked by non-nucleotide components. In this application, the terms "nucleic acid" and "polynucleotide" can be double-stranded and single-stranded molecules. Unless otherwise specified or required, any embodiment of the polynucleotide in this application includes each of the double-stranded form and the two complementary single-stranded forms known or predicted to constitute the double-stranded form. For example, a polynucleotide can include, but is not limited to, a gene or gene fragment (e.g., a probe, primer, EST or SAGE tag), exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, siRNA, miRNA, shRNA, RNAi agents, and primers.

[0169] In this application, the term "oligonucleotide" generally refers to a polymer composed of a plurality of nucleotide residues (deoxyribonucleotides or ribonucleotides, or their relevant structural variants or synthetic analogs) connected by a phosphodiester bond (or their relevant structural variants or synthetic analogs). Therefore, although the term "oligonucleotide" generally refers to a naturally occurring nucleotide polymer in which the nucleotide residues and the connection therebetween are naturally occurring, it should be understood that the scope of the term also includes various analogs, including but not limited to: peptide nucleic acids (PNAs), phosphoramidates, phosphorothioates, methylphosphonates, 2-O-methylribonucleic acids, etc. The exact size of the molecule can depend on specific applications. Oligonucleotides are generally shorter in length, typically having approximately 10-30 nucleotide residues, but the term can also refer to molecules of any length, although the terms "polynucleotides" or "nucleic acids" are generally used for larger oligonucleotides.

[0170] In certain embodiments, the oligonucleotide comprises one or more unmodified ribonucleosides (RNA) and / or unmodified deoxyribonucleosides (DNA) and / or one or more modified nucleosides. The term "modified oligonucleotide" generally refers to an oligonucleotide comprising at least one modified nucleoside and / or at least one modified internucleoside linkage.

[0171] In this application, the term "modified nucleoside" generally means a nucleoside that contains at least one chemical modification compared to naturally occurring RNA or DNA nucleosides. The modified nucleoside contains a modified sugar moiety and / or a modified nucleobase.

[0172] In this application, the term "nucleobase" generally refers to a heterocyclic pyrimidine or purine compound, which is a component of all nucleic acids and includes adenine (a), guanine (g), cytosine (c), thymine (t) and uracil (u). Nucleotides may include modified nucleotides or nucleotide mimetics, abasic sites (Ab or X) or substitute replacement moieties. As used herein, a "nucleobase sequence" generally refers to the order of consecutive nucleobases that are independent of any sugar, linkage or nucleobase modification. The term "unmodified nucleobase" or "naturally occurring nucleobase" generally refers to the naturally occurring heterocyclic nucleobases of RNA or DNA: the purine bases adenine (a) and guanine (g); and thymine (t), cytosine (c) (including 5-methyl c) and uracil (u). A "modified nucleobase" generally refers to any nucleobase that is not a naturally occurring nucleobase.

[0173] In this application, the term "sugar moiety" generally refers to a naturally occurring sugar moiety or a modified sugar moiety of a nucleoside. The term "naturally occurring sugar moiety" generally refers to a ribofuranosyl group as found in naturally occurring RNA or a deoxyribofuranosyl group as found in naturally occurring DNA. A "modified sugar moiety" refers to a substituted sugar moiety or a sugar surrogate.

[0174] In this application, the term "internucleoside linkage" generally refers to the covalent linkage between adjacent nucleosides in an oligonucleotide. "Naturally occurring internucleoside linkage" means a 3' to 5' phosphodiester linkage. "Modified internucleoside linkage" means any internucleoside linkage other than a naturally occurring internucleoside linkage.

[0175] As used herein, the term "target nucleic acid" or "target sequence" generally refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during transcription of the IGF-1R gene, including mRNAs that are RNA processing products of the primary transcription product. The target portion of the sequence should be at least long enough to serve as a substrate for iRNA-directed cleavage at or near that portion of the nucleotide sequence of the mRNA molecule formed during transcription of the IGF-1R gene. In one embodiment, the target sequence is within the protein-coding region of IGF-1R. The target sequence can be approximately 19-36 nucleotides in length, for example, preferably approximately 19-30 nucleotides in length. Ranges and lengths intermediate to the aforementioned ranges and lengths are also intended to be included as part of this application.

[0176] As used herein, the term "IGF-1R protein" refers to a human tetrameric transmembrane tyrosine kinase. The IGF-1R protein is a cell surface receptor for the hormone insulin-like growth factor 1, a hormone similar in molecular structure to the polypeptide hormone insulin, and plays an important role in growth and development and adult anabolism. Activation of IGF-1R can stimulate cell proliferation, survival, transformation, metastasis, and angiogenesis. As used herein, the term "IGF-1R gene" refers to the gene encoding the IGF-1R protein, which has a molecular weight of approximately 320 kDa and is composed of two α subunits and two β subunits. The IGF-1R gene can be regulated or inhibited by RNA inhibitors. In some embodiments, the RNA inhibitors can regulate or inhibit the mRNA sequence of the IGF-1R gene (NM_000875.5 Homo sapiens insulin-like growth factor 1 receptor). The transcript sequence of the IGF-1R gene is shown in SEQ ID NO: 828.

[0177] In the present application, term " blocking group " refers to the group that can selectively be combined with duplex siRNA described in the present application or the drug that comprises duplex siRNA, and described blocking group is positioned at sense strand or sense nucleic acid fragment, antisense strand or antisense nucleic acid fragment or two chains.Blocking group can be attached to 3 ' end, 5 ' end or two ends of sense strand or sense nucleic acid fragment, antisense strand or antisense nucleic acid fragment or two chains.In certain embodiments, blocking group is incorporated into antisense strand or antisense nucleic acid fragment, particularly 5 ' end of antisense strand or antisense nucleic acid fragment.In certain embodiments, blocking group is combined with oligonucleotide (for example, 5 ' end of antisense strand or antisense nucleic acid fragment) by internucleotide link, and this internucleotide link is optionally modified as described above.In certain embodiments, blocking group is linked to double-stranded oligonucleotide medicament by phosphosulfate.In certain embodiments, blocking group is linked to double-stranded oligonucleotide medicament by phosphodiester. In the present application, in order to reduce or inhibit the degradation of exonucleases, a "blocking group" may be incorporated into the 5' end of the duplex siRNA antisense strand or antisense nucleic acid fragment described herein. In certain embodiments, the blocking group may reduce or inhibit the RNA interference effect of the oligonucleotide. In certain embodiments, the blocking group is excised from the oligonucleotide before providing the RNA interference effect. For example, examples of blocking groups include, but are not limited to, abasic residues, reverse abasic residues, and (M06).

[0178] In this application, the term "ligand" generally refers to any compound or molecule that can be covalently or otherwise chemically bound to a biologically active substance (such as an oligonucleotide). In certain embodiments, a ligand can interact directly or indirectly with another compound, such as a receptor. The receptor that interacts with the ligand can be present on the cell surface, or alternatively can be an intracellular and / or intercellular receptor. The interaction of the ligand with the receptor can result in a biochemical reaction, or can be simply a physical interaction or binding.

[0179] In the present application, "conjugation" refers to the connection between two or more chemical moieties, each of which has a specific function, in a covalently linked manner; accordingly, "conjugate" refers to a compound formed by covalent linkage between the various chemical moieties. For example, "double-stranded RNA conjugate" means a compound or complex formed by covalently linking one or more chemical moieties (such as a delivery system, a ligand group, or a conjugated group) with a specific function to a double-stranded RNA. In some embodiments, the delivery system, ligand group, or conjugated group can be connected to the phosphate group, sugar ring (including the delivery system, ligand group, or conjugated group being covalently linked to the atom at the 3' or 5' position of the nucleotide via a phosphodiester bond), 2'-hydroxyl group, 5'-hydroxyl group, or base of any nucleotide of the double-stranded RNA. In some embodiments, the delivery system, ligand group, or conjugated group can also be connected to the 2'-position of the nucleotide, where the nucleotides are linked using a 2'-5' phosphodiester bond. In some embodiments, the delivery system, ligand group, or conjugate group can also be attached to the 3' position of the nucleotide, in which case the nucleotides are connected using a 3'-5' phosphodiester bond. In some embodiments, the connection between the delivery system, ligand group, or conjugate group and the nucleotide can be further modified, such as thio modification (i.e., connection via a thiophosphodiester bond).

[0180] In this application, the terms "induce," "inhibit," "enhance," "elevate," "increase," "reduce," "reduced," and the like generally refer to a quantitative difference between two states. For example, "an amount effective to inhibit the activity or gene expression of IGF-1R" means that the level of IGF-1R activity or gene expression in a treated sample will be lower than the level of IGF-1R activity or gene expression in an untreated sample. Such terms apply, for example, to gene expression levels and activity levels. The terms "reduce" and "reduce" are used interchangeably and generally refer to any change that is less than the original value. "Reduce" and "reduce" are relative terms, requiring a comparison between before and after a measurement. "Reduce" and "reduce" include complete depletion.

[0181] In certain embodiments, the term "reduce" refers to a gene or biomarker gene expression level / amount detected by standard methods known in the art (such as those described in this application), a gene, a gene product, such as a protein or a biomarker in a first sample, and a gene expression level / amount of a corresponding gene, a gene product, such as a protein or a biomarker in a second sample, compared to about 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95% or 100% of the overall reduction. In certain embodiments, the term "reduce" refers to a reduction in the gene expression level / amount of a gene or biomarker in a first sample, wherein the reduction is at least about 0.9 times, 0.8 times, 0.7 times, 0.6 times, 0.5 times, 0.4 times, 0.3 times, 0.2 times, 0.1 times, 0.05 times or 0.01 times of the gene expression level / amount of the corresponding gene or biomarker in the second sample. In certain embodiments, the first sample is a sample obtained from a subject, and the second sample is a reference sample.

[0182] In this application, the term "gene expression" generally refers to the process by which a gene ultimately produces a protein, including but not limited to transcription, post-transcriptional modification (eg, splicing, polyadenylation, addition of a 5'-cap), and translation.

[0183] In this application, the term "pharmaceutically acceptable" generally refers to one or more non-toxic substances that do not interfere with the effectiveness of the biological activity of the active ingredient. Such formulations may generally contain salts, excipients, buffers, preservatives, compatible carriers, and optionally other therapeutic agents. Such pharmaceutically acceptable formulations may also generally include compatible solid or liquid fillers, diluents, or encapsulating materials suitable for administration to humans. When used in medicine, the salt should be a pharmaceutically acceptable salt, but non-pharmaceutically acceptable salts can be conveniently used to prepare pharmaceutically acceptable salts, and these are not excluded from the scope of this application. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, salts prepared from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, citric acid, boric acid, formic acid, malonic acid, succinic acid, and the like. Pharmaceutically acceptable salts may also be prepared as alkali metal or alkaline earth metal salts, such as sodium, potassium, or calcium salts.

[0184] In this application, the term "prevention and / or treatment" includes not only preventing and / or treating a disease, but also generally includes preventing the onset of a disease, slowing or reversing the progression of a disease, preventing or slowing the onset of one or more symptoms associated with the disease, reducing and / or alleviating one or more symptoms associated with the disease, reducing the severity and / or duration of the disease and / or any symptoms associated therewith and / or preventing further increase in the severity of the disease and / or any symptoms associated therewith, preventing, reducing or reversing any physiological damage caused by the disease, and generally any pharmacological effect that is beneficial to the patient being treated. The RNAi agent or pharmaceutical composition of the present application does not need to achieve a complete cure or eradication of any symptoms or manifestations of the disease to form a viable therapeutic agent. As recognized in the relevant art, drugs used as therapeutic agents can reduce the severity of a given disease state, but do not need to eliminate every manifestation of the disease to be considered a useful therapeutic agent. Similarly, a prophylactic treatment does not need to be completely effective in preventing the onset of the disease. Simply reducing the impact of the disease in the subject (e.g., by reducing the number or severity of its symptoms, or by increasing the effectiveness of another treatment, or by producing another beneficial effect) or reducing the likelihood of the disease occurring or worsening is sufficient.

[0185] In this application, the terms "disease" or "disorder" are used interchangeably and generally refer to any deviation of a subject from the normal state, such as any change in the state of the body or certain organs that prevents or disrupts the performance of functions and / or causes symptoms such as discomfort, dysfunction, suffering or even death in the person suffering from the disease or contact therewith. Disease or disorder may also be referred to as distemper, discomfort, ailment, malady, disorder, sickness, illness, complaint.

[0186] In this application, the term "administer" generally refers to introducing the present pharmaceutical preparation into the body of a subject by any introduction or delivery route. Any method known to those skilled in the art for contacting cells, organs or tissues with the drug can be used. The administration may include, but is not limited to, intravenous, intraarterial, intranasal, intraperitoneal, intramuscular, subcutaneous transdermal or oral administration. The daily dose may be divided into one, two or more suitable forms of dosage to be administered at one, two or more times during a certain time period.

[0187] In the present application, the term "contact" generally refers to two or more different types of substances being contacted together in any order, in any manner, and for any duration. Contact can occur in vivo, ex vivo, or in vitro. In certain embodiments, it may refer to making the RNAi agent or composition of the present application directly contact cells or tissues. In other embodiments, the term refers to making the RNA inhibitor or composition of the present application indirectly contact cells or tissues. For example, the method of the present application includes a method in which a subject contacts an RNA inhibitor or composition of the present application, and then the RNA inhibitor or composition contacts cells or tissues by diffusion or any other active transport or passive transport process known in the art (the compound circulates in the body through this process).

[0188] In this application, the term "effective amount" or "effective dose" generally refers to an amount sufficient to achieve or at least partially achieve the desired effect. A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is generally any amount of the drug that, when used alone or in combination with another therapeutic agent, promotes disease regression (as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or the prevention of damage or disability caused by the disease). A "prophylactically effective amount" or "prophylactically effective dose" of a drug generally refers to an amount of the drug that, when administered alone or in combination with another therapeutic agent to a subject at risk of disease progression or disease recurrence, inhibits the progression or recurrence of the disease. The ability of a therapeutic or prophylactic agent to promote disease regression or inhibit disease progression or recurrence can be assessed using a variety of methods known to those skilled in the art, such as in human subjects during clinical trials, predicting efficacy in humans in animal model systems, or by measuring the activity of the agent in in vitro assays. In certain embodiments, an "effective amount" refers to the amount of an RNA inhibitor that produces the desired pharmacological, therapeutic or prophylactic result.

[0189] In this application, the term "subject" generally refers to a human or non-human animal (including mammals) in need of diagnosis, prognosis, improvement, prevention and / or treatment of a disease, such as humans, non-human primates (apes, gibbons, gorillas, chimpanzees, orangutans, macaques), livestock (dogs and cats), farm animals (poultry such as chickens, ducks, horses, cattle, goats, sheep, pigs) and experimental animals (mice, rats, rabbits, guinea pigs). Human subjects include fetuses, newborns, infants, adolescents and adult subjects. Subjects include animal disease models.

[0190] In this application, the terms "include," "comprising," "having," "may," "containing," and variations thereof are generally intended to be open transitional phrases, terms, or words that do not exclude the possibility of additional actions or structures. The term "consisting of" generally indicates that no other components (or, similarly, features, integers, steps, etc.) can be present. Singular forms such as "a," "an," and "the" in English, and "one," "a," and "said / the" in Chinese generally include plural forms of the referent unless the context clearly dictates otherwise.

[0191] In this application, the term "about" generally means approximately, roughly, or around. When the term "about" is used in reference to a numerical range, a cutoff or specific value is used to indicate that the stated value may vary from the recited value by up to 10%. Thus, the term "about" can be used to encompass variations of ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from the specified value.

[0192] It should be understood that the term "at least" preceding a number or a range of numbers includes the number adjacent to the term "at least" and all subsequent numbers or integers that are logically included, as clear from the context. For example, the number of nucleotides in a nucleic acid molecule must be an integer. For example, "at least 19 nucleotides in a 21-nucleotide nucleic acid molecule" means that 19, 20, or 21 nucleotides have the indicated property. When "at least" appears before a series of numbers or a range, it should be understood that "at least" can modify each number in the series or range.

[0193] It should be understood that "not more than" or "less than" as used herein refers to the value or integer that is adjacent to the phrase and is logically lower, such as zero, as the context makes logical sense. For example, a duplex having an overhang of "not more than 3 nucleotides" has an overhang of 3, 2, 1, or 0 nucleotides. When "not more than" appears before a series of numbers or a range, it should be understood that "not more than" can modify each number in the series or range. As used herein, ranges include both upper and lower limits.

[0194] Detailed Description of the Invention

[0195] Antisense strand or antisense nucleic acid fragment and sense strand or sense nucleic acid fragment

[0196] In one aspect, the present application provides an RNA inhibitor for inhibiting insulin-like growth factor 1 receptor (IGF-1R) gene expression, comprising an antisense strand, wherein the antisense strand forms a complementary region with at least 15 consecutive nucleotides in a sequence encoding IGF-1R (SEQ ID NO: 828), wherein the complementary region has 0, 1, 2, 3, 4, or 5 mismatches, and preferably the duplex complementary region is 15-30 nucleotide pairs in length, more preferably 17-23 nucleotide pairs in length. For example, the duplex complementary region can be 17 nucleotide pairs, 18 nucleotide pairs, 19 nucleotide pairs, 20 nucleotide pairs, 21 nucleotide pairs, 22 nucleotide pairs, or 23 nucleotide pairs in length.

[0197] In certain embodiments, the RNA inhibitor comprises a single-stranded oligonucleotide or double-stranded ribonucleic acid (dsRNA) molecule for inhibiting expression of an IGF-1R gene in a cell, such as a cell of a subject (e.g., a mammal), wherein the dsRNA comprises an antisense strand or antisense nucleic acid fragment having a complementary region that is complementary to at least a portion of an mRNA formed during expression of the IGF-1R gene, wherein the complementary region is about 12-30 nucleotides in length (e.g., about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, or 12 nucleotides in length).

[0198] dsRNA comprises two RNA chains, which can complement and hybridize to form a duplex structure (complementary region) under the conditions of dsRNA use. One chain of dsRNA (the antisense strand or antisense nucleic acid fragment) includes a complementary region that is substantially complementary and usually completely complementary to the antisense strand. It can be derived from the sequence of mRNA formed during IGF-1R gene expression. The other chain (the sense strand or sense nucleic acid fragment) includes a region complementary to the antisense strand or antisense nucleic acid fragment, so that when combined under appropriate conditions, the two chains can hybridize and form a duplex structure. Typically, the length of the duplex structure is 12 to 30 base pairs. Similarly, the complementary region to is 12 to 30 nucleotides in length, for example, at 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-2 27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length.

[0199] In certain embodiments, the dsRNA is about 19 to about 23 nucleotides in length, or about 24 to about 30 nucleotides in length. Typically, the length of the dsRNA is sufficient to serve as a substrate for the Dicer enzyme. For example, it is well known in the art that dsRNAs greater than about 21-23 nucleotides in length can be used as substrates for Dicer. It is also understood by those skilled in the art that the region of the RNA targeted for cleavage is typically a portion of a larger RNA molecule (typically an mRNA molecule). A "portion" of a target is a continuous nucleotide of an mRNA target that is long enough to allow it to be a substrate for RNAi-guided cleavage (i.e., cleavage via the RISC pathway).

[0200] It will also be understood by those skilled in the art that the region of complementarity is the major functional portion of a dsRNA, e.g., the duplex region of about 19 to about 30 base pairs, e.g., about 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21 , 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs.

[0201] In certain embodiments, there is at least about 80% base complementarity between the sense strand or sense nucleic acid segment and the antisense strand or antisense nucleic acid segment.

[0202] In certain embodiments, the sense strand or sense nucleic acid fragment and the antisense strand or antisense nucleic acid fragment are each independently 19-30 nucleotides.

[0203] In certain embodiments, the sense strand or sense nucleic acid fragment and the antisense strand or antisense nucleic acid fragment are each independently 17-25 nucleotides.

[0204] In certain embodiments, the sense strand or sense nucleic acid fragment and the antisense strand or antisense nucleic acid fragment are each independently 19-23 nucleotides.

[0205] In some embodiments, the total length of the sense strand is 15-30 nucleotides, preferably the total length of the sense strand is 16-23 nucleotides, and more preferably the total length of the sense strand is 19, 20 or 21 nucleotides.

[0206] In some embodiments, the total length of the antisense strand is 19-30 nucleotides, preferably the total length of the antisense strand is 19-27 nucleotides, and more preferably the total length of the antisense strand is 21, 22, 23, 24, 25, 26 or 27 nucleotides.

[0207] In some embodiments, the sense strand or sense nucleic acid fragment is selected from any one of SEQ ID NOs: 1-198 or a sequence that differs therefrom by no more than 3 nucleotides.

[0208] In some embodiments, the sense strand or sense nucleic acid fragment of the RNA inhibitor is selected from sequences in Table 1 that differ by one, two, or three nucleotides, such as 15, 16, 17, 18, 19, or 20 consecutive nucleotides.

[0209] In some embodiments, the antisense strand or antisense nucleic acid fragment is selected from any one of SEQ ID NOs: 199-413 or a sequence of at least 15 consecutive nucleotides that differs therefrom by no more than 3 nucleotides, such as 15, 16, 17, 18, 19 or 20 consecutive nucleotides.

[0210] In some embodiments, the antisense strand or antisense nucleic acid fragment of the RNA inhibitor is selected from at least 15 consecutive nucleotide sequences that differ from each sequence in Table 1 by one, two, or three nucleotides, such as 15, 16, 17, 18, 19, or 20 consecutive nucleotides.

[0211] In some embodiments, the sense and antisense strands each independently optionally comprise a 3' or 5' overhang of 1, 2, or 3 nucleotides.

[0212] In some embodiments, both the sense strand and the antisense strand have 3' overhangs of 1-3 nucleotides in length, or the sense strand has a 3' or 5' overhang of 1-3 nucleotides in length, or the antisense strand has a 3' or 5' overhang of 1-3 nucleotides in length. In some embodiments, the RNA inhibitors described herein have specific structures near their 5' and 3' ends, as shown in Figure 2.

[0213] In certain embodiments, the antisense strand comprises a cleavage region comprising a nucleotide sequence represented by formula (I),

[0214] Formula (I): (3'-5')X2-YZ,

[0215] The cleavage occurs between X2 and Y, wherein X2 is the 5'-most nucleotide of the second chain, Y and Z are the two 3'-most nucleotides of the 5' extension, and Z is guanine nucleotide (G), a natural analog of guanine nucleotide (G), a non-natural analog of guanine nucleotide (G), adenine nucleotide (A), a natural analog of adenine nucleotide (A), or a non-natural analog of adenine nucleotide (A).

[0216] In certain embodiments, Z is guanine nucleotide (G), a natural analog of guanine nucleotide (G), or a non-natural analog of guanine nucleotide (G).

[0217] In certain embodiments, X2 is adenine nucleotide (A), a natural analogue of adenine nucleotide (A), a non-natural analogue of adenine nucleotide (A), uracil nucleotide (U), a natural analogue of uracil nucleotide (U), or a non-natural analogue of uracil nucleotide (U).

[0218] In certain embodiments, wherein the formula (I) has a sequence (3'-5') selected from the group consisting of UUG, UAG, AUG, AAG, UUA, UAA, AUA, AAA, UCG, UGG, ACG, AGG, UCA, UGA, ACA and AGA, or a natural or non-natural analogue thereof.

[0219] In certain embodiments, Y is an adenine nucleotide (A), a natural analog of an adenine nucleotide (A), a non-natural analog of an adenine nucleotide (A), a uracil nucleotide (U), a natural analog of a uracil nucleotide (U), or a non-natural analog of a uracil nucleotide (U).

[0220] In certain embodiments, the formula (I) has a sequence (3'-5') selected from the group consisting of UUG, UAG, AUG, AAG, UUA, UAA, AUA, and AAA.

[0221] In certain embodiments, the cleavage region further comprises nucleotide N1, wherein N1 is the third nucleotide from the 3' end of the 5' extension, and the cleavage region comprises the nucleotide sequence shown in formula (II),

[0222] Formula (II): (3'-5')X2-YZ-N1.

[0223] In certain embodiments, the cleavage region further comprises a fragment N, wherein the fragment N comprises at least one nucleotide, wherein the nucleotide at the 3' end of the fragment N is N1, and the cleavage region comprises a nucleotide sequence represented by formula (III),

[0224] Formula (III): (3'-5')X2-YZN,

[0225] The length of the fragment N is 1-10 nucleotides, preferably 1-5 nucleotides, and more preferably 1 nucleotide.

[0226] In certain embodiments, N is an adenine nucleotide (A), a guanine nucleotide (G), a cytosine nucleotide (C), a uracil nucleotide (U), a natural analogue thereof, or a non-natural analogue thereof; preferably, N1 is a cytosine nucleotide (C), a natural analogue of a cytosine nucleotide (C), or a non-natural analogue of a cytosine nucleotide (C).

[0227] In certain embodiments, the formula (III) has a sequence (3'-5') selected from the group consisting of AAGC and UAGC.

[0228] In some embodiments, the sense strand or sense nucleic acid fragment and antisense strand of the RNA inhibitor are selected from at least 15 consecutive nucleotide sequences that differ by one, two, or three nucleotides from each sequence in Table 1, such as 15, 16, 17, 18, 19, or 20 consecutive nucleotides.

[0229] Table 1 siRNA sense and antisense strand sequences

[0230] Modified nucleotides

[0231] To enhance the in vivo stability of the RNA inhibitor, the sense and antisense strands of the RNA inhibitor can be modified without affecting or even enhancing their activity. The nucleotides can have modifying groups, and the entire strand or a portion thereof can be modified. In certain embodiments, one or more nucleotides in the sense and / or antisense strands are modified to form modified nucleotides.

[0232] As used herein, the term "nucleotide" refers to a 5-carbon sugar (ribose or deoxyribose), a phosphate group, and a base (natural or non-natural base), and is intended to include unmodified (i.e., natural) nucleotides and modified nucleotides. In certain embodiments, the nucleotide is an unmodified ribonucleotide. In certain embodiments, the ribonucleotide is a 3'-ribonucleotide. In certain embodiments, the ribonucleotide is a 5'-ribonucleotide. In certain embodiments, the modified or unmodified nucleotide may optionally be further modified.

[0233] Natural nucleotides are composed of natural bases, natural ribose, and phosphate. As used herein, natural nucleotides refer to adenine ribonucleotides, adenine deoxyribonucleotides, guanine ribonucleotides, guanine deoxyribonucleotides, cytosine ribonucleotides, cytosine deoxyribonucleotides, uracil ribonucleotides, thymine ribonucleotides, or thymine deoxyribonucleotides. "Ribonucleotides" refer to nucleotides with a hydroxyl group at the 2' position of the sugar portion of the nucleotide. "Deoxyribonucleosides" refer to nucleotides with a hydrogen group at the 2' position of the sugar portion of the nucleotide.

[0234] The natural bases of RNA include A (adenine), G (guanine), C (cytosine), U (uracil), and T (thymine).

[0235] As used herein, the mark "d" before a monomer (eg, nucleotides A, U, C, G, and T, etc.) indicates that the monomer is modified by 2'-deoxy.

[0236] As used herein, the mark "f" after a monomer (such as nucleotides A, U, C, G, and T, etc.) indicates that the monomer is modified with 2'-deoxy-2'-fluoro (2'-F modification).

[0237] As used herein, the mark "GNA-" before a monomer (such as nucleotides A, U, C, G, and T) indicates that the monomer is modified with ethylene glycol nucleic acid (GNA modification). As used herein, Tgn is the code for GNA-T, which is equivalent.

[0238] As used herein, lowercase letters (a, u, c, g, t, etc.) indicate that the nucleotide represented by its corresponding uppercase letter (A, U, C, G, and T, etc.) is modified with a 2'-O-methyl group (2'-OMe).

[0239] As used herein, the invAB modification refers to the inverted abasic nucleotides bonded to the 5' or 3' end of the nucleotide. The structure after modification by invAB:

[0240] As used herein, M06 modification refers to the addition of a 5' or 3' end bond to a monomer (such as a nucleotide). For example, The structure after being modified by M06 through phosphate bonds: In the present invention, (M06) It is an M06 monomer In some embodiments, (M06) is represented by Bonded to a nucleotide.

[0241] As used herein, the mark "*" between monomers (such as nucleotides A, U, C, G and T, etc.) indicates that the two monomers are connected by a phosphorothioate bond (i.e., phosphorothioate diester bond), i.e., modified by phosphorothioate (PS).

[0242] As used herein, the absence of a "*" mark between nucleotides (A, U, C, G, T, etc.) indicates that the two nucleotides are linked by a phosphate bond (ie, a phosphodiester bond).

[0243] For example, "5'-Phosphorothioate-invAB" means that (invAB) is linked to the 5' end of the monomer via a phosphorothioate bond. For example, "5'-Phosphorothioate-M06" means that (M06) is linked to the 5' end of the monomer via a phosphorothioate bond.

[0244] All nucleotides in the small inhibitory nucleic acid molecule described herein may be natural or unmodified nucleotides, or at least one nucleotide may be a modified nucleotide, wherein the modification is one or a combination of the following modifications:

[0245] (1) modification of the phosphodiester bonds of the nucleotides in the nucleotide sequence of the small inhibitory nucleic acid molecule;

[0246] (2) modification of the 2'-OH group of ribose in the nucleotide sequence of the small inhibitory nucleic acid molecule;

[0247] (3) Modification of bases in the nucleotide sequence of the small inhibitory nucleic acid molecule.

[0248] Chemical modifications are well known to those skilled in the art. Phosphodiester modifications refer to modifications of the oxygen in the phosphodiester bond, including phosphorothioate and phosphate borylation. Both modifications stabilize the siRNA structure and maintain high base pairing specificity and affinity.

[0249] The ribose modification refers to the modification of the 2'-OH in the pentose of the nucleotide, that is, the introduction of certain substituents at the hydroxyl position of the ribose, for example, 2'-fluoro modification, 2'-oxymethyl modification, 2'-oxyethylenemethoxy modification, 2,4'-dinitrophenol modification, locked nucleic acid (LNA), 2'-amino modification, 2'-deoxy modification.

[0250] The base modification refers to modification of the base of the nucleotide, for example, 5'-bromouracil modification, 5'-iodouracil modification, N-methyluracil modification, and 2,6-diaminopurine modification.

[0251] In some embodiments, the modification of the ribose comprises fluorine substitution and / or methoxy substitution on the 2'-OH.

[0252] In some embodiments, the modification of the ribose further comprises modification by UNA, LNA or GNA.

[0253] The term "LNA" refers to bicyclic nucleoside analogs containing a C2*-C4* diradical (bridge) and is referred to as "locked nucleic acid." It can refer to an LNA monomer or, when used in the context of an "LNA oligonucleotide," an oligonucleotide containing one or more such bicyclic nucleotide analogs. In certain aspects, bicyclic nucleoside analogs are LNA nucleotides, and these terms are therefore used interchangeably, and in such embodiments, both are characterized by the presence of a linker (such as a bridge) between the C2' and C4' ribose sugar rings.

[0254] UNA (unlocked nucleic acid) has a structure similar to RNA, but lacks the C2 and C3 chemical bonds of the ribose ring. Its structure is shown below:

[0255] Where B is a base.

[0256] Glycerol nucleic acid (GNA) is a chemical substance similar to DNA or RNA, but its composition differs and it does not exist naturally in any known organism. GNA contains an acyclic three-carbon propylene glycol (1,2-propylene glycol) backbone, which replaces the (deoxy)ribose sugars of DNA and RNA, forming the simplest chemically stable nucleic acid structure. The structure of S-(GNA) is shown below.

[0257] Where B is a base.

[0258] In certain embodiments, the RNA inhibitor, wherein at least one nucleotide in the RNA inhibitor is a chemically modified nucleotide.

[0259] In certain embodiments, the RNA inhibitor, wherein all nucleotides in the RNA inhibitor are chemically modified nucleotides.

[0260] In certain embodiments, the RNA inhibitor, wherein the modification comprises a combination of one or more of the following: 2'-OMe (2'-O-methyl) modification, 2'-F (2'-deoxy-2'-fluoro) modification, 2'-O-MOE (2'-O-methoxyethyl) modification, 2'-deoxy (2'-d) modification, 5'-morpholine (5'-Mo) modification, unlocked nucleic acid (UNA) modification, glycol nucleic acid (GNA) modification, locked nucleic acid (LNA) modification, tricyclic DNA (tcDNA) modification, (S)-constrained ethyl bicyclic nucleic acid ((S)-cEt-BNA) modification, phosphorothioate (PS) modification, phosphorodithioate (PS2) modification, methylphosphonate (MP) modification, methoxypropylmethylphosphonate (MOP) modification, peptide nucleic acid (PNA) modification, 5'-(E)-vinyl phosphate (VP) modification. modification (VP), N6-methyladenosine (m6A) modification, 5-methylcytidine (m5C) modification, 3-methyluridine (m3U) modification, 5-methyluridine (m5U) modification, pseudouridine modification, 2-thiouridine (s2U) modification, propyne uridine (5-pU) modification, inverted abasic nucleotide (invAB) modification by bonding the 5' or 3' end of the nucleotide, replacing the nucleotide with an inverted abasic nucleotide (invAb) modification, replacing the nucleotide with a 2,4-difluoromethylphenyl ribonucleotide (rF) modification or replacing the nucleotide with a (S)-glycerol nucleic acid modification, preferably 2'-OMe modification, 2'-F modification, 2'-deoxy modification, VP modification, 5'-MP modification, PS modification, PS2 modification, MP modification, MOP modification, invAb modification, invAB modification.

[0261] In certain embodiments, in the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 5' end, the 9th nucleotide of the sense strand has a 2'-F modification.

[0262] In certain embodiments, in the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 5' end, the 10th nucleotide of the sense strand has a 2'-F modification.

[0263] In certain embodiments, in the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 5' end, the 11th nucleotide of the sense strand has a 2'-F modification.

[0264] In certain embodiments, when the sense strand of the RNA inhibitor is 21 nt in length, counting from the 5' end, the 9th, 10th, and 11th nucleotides of the sense strand are modified with 2'-F, and the rest are modified with 2'-OMe.

[0265] In certain embodiments, the RNA inhibitor, when the sense strand is 21 nt in length, counting starts from the 5' end, wherein the 5' end of the first nucleotide of the sense strand is modified with invAB.

[0266] In certain embodiments, in the RNA inhibitor, the inverted abasic nucleotide is linked to the 5' end of the first nucleotide of the sense strand via a phosphorothioate.

[0267] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a phosphorothioate linkage between the first and second nucleotides of the antisense strand.

[0268] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a phosphorothioate linkage between the second and third nucleotides of the antisense strand.

[0269] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a phosphorothioate linkage between the 3rd and 4th nucleotides of the antisense strand.

[0270] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 3' end, has a phosphorothioate linkage between the first and second nucleotides of the antisense strand.

[0271] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the antisense strand, a phosphorothioate linkage between the 2nd and 3rd nucleotides, and a phosphorothioate linkage between the 3rd and 4th nucleotides; and counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the antisense strand.

[0272] In certain embodiments, in the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, the second nucleotide of the antisense strand has a 2'-F modification.

[0273] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a 2'-F modification at the third nucleotide of the antisense strand.

[0274] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a 2'-F modification at the 4th nucleotide of the antisense strand.

[0275] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a 2'-F modification at the 12th nucleotide of the antisense strand.

[0276] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a 2'-F modification at the 14th nucleotide of the antisense strand.

[0277] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, has a 2'-F modification at the 16th nucleotide of the antisense strand.

[0278] In certain embodiments, when the antisense strand of the RNA inhibitor is 21 nt in length, counting from the 5' end, the nucleotides at positions 2, 3, 4, 12, 14, and 16 of the antisense strand are modified with 2'-F, and the rest are modified with 2'-OMe.

[0279] In certain embodiments, the RNA inhibitor, when the antisense strand is 21 nt in length, counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the antisense strand, a phosphorothioate linkage between the 2nd and 3rd nucleotides, and a phosphorothioate linkage between the 3rd and 4th nucleotides; and counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the antisense strand; and nucleotides at positions 2, 3, 4, 12, 14, and 16 of the antisense strand are 2'-F modified, and the rest are 2'-OMe modified.

[0280] In certain embodiments, the RNA inhibitor,

[0281] a) When the sense strand is 21 nt in length, the sense strand includes the following modifications: counting from the 5' end, the 9th, 10th, and 11th nucleotides have 2'-F modifications, and the rest are 2'-OMe modifications; counting from the 5' end, the 5' end of the first nucleotide has an invAB modification,

[0282] b) When the antisense strand is 21 nt in length, the antisense strand comprises the following modifications: counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, a phosphorothioate linkage between the 2nd and 3rd nucleotides, and a phosphorothioate linkage between the 3rd and 4th nucleotides; and counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides; and there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16, and the rest are 2'-OMe modifications.

[0283] In certain embodiments, the RNA inhibitor, when the sense strand is 21 nt in length, counted from the 5' end, has a 2'-F modification at the 18th nucleotide.

[0284] In certain embodiments, when the sense strand of the RNA inhibitor is 21 nt in length, counting from the 5' end, the 9th, 10th, 11th, and 18th nucleotides are modified with 2'-F, and the rest are modified with 2'-OMe.

[0285] In certain embodiments, the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 5' end, has a phosphorothioate linkage between the first and second nucleotides.

[0286] In certain embodiments, the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 5' end, has a phosphorothioate linkage between the 2nd and 3rd nucleotides.

[0287] In certain embodiments, the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 3' end, has a phosphorothioate linkage between the first and second nucleotides.

[0288] In certain embodiments, the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 3' end, has a phosphorothioate linkage between the 2nd and 3rd nucleotides.

[0289] In certain embodiments, the RNA inhibitor, when the sense strand is 21 nt in length, counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, a phosphorothioate linkage between the 2nd and 3rd nucleotides, 2'-F modifications are present at nucleotides 9, 10, 11, and 18 of the sense strand, and the rest are 2'-OMe modified; counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, and a phosphorothioate linkage between the 2nd and 3rd nucleotides of the sense strand.

[0290] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 5' end, has a 2'-F modification at the first nucleotide of the antisense strand.

[0291] In certain embodiments, in the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 5' end, the second nucleotide of the antisense strand has a 2'-F modification.

[0292] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 5' end, has a 2'-F modification at the 5th nucleotide of the antisense strand.

[0293] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 5' end, has a 2'-F modification at the 9th nucleotide of the antisense strand.

[0294] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 5' end, has a 2'-F modification at the 17th nucleotide of the antisense strand.

[0295] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 5' end, has a 2'-F modification at the 19th nucleotide of the antisense strand.

[0296] In certain embodiments, when the antisense strand is 26 nt in length, counting from the 5' end, the nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense strand are modified with 2'-F, and the rest are modified with 2'-OMe.

[0297] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 5' end, has a phosphorothioate linkage between the 4th and 5th nucleotides.

[0298] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 3' end, has a phosphorothioate linkage between the 1st and 2nd nucleotides.

[0299] In certain embodiments, the RNA inhibitor, when the antisense strand is 26 nt in length, counting from the 3' end, has a phosphorothioate linkage between the 2nd and 3rd nucleotides.

[0300] In certain embodiments, when the antisense strand of the RNA inhibitor is 26 nt in length, counting from the 5' end, nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense strand are modified with 2'-F, and the rest are modified with 2'-OMe, and there is a phosphorothioate linkage between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides.

[0301] In certain embodiments, the RNA inhibitor, wherein

[0302] a) When the sense strand is 21 nt in length, counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, a phosphorothioate linkage between the 2nd and 3rd nucleotides, 2'-F modifications are present at nucleotides 9, 10, 11, and 18 of the sense strand, and the remaining nucleotides are 2'-OMe modified; counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, and a phosphorothioate linkage between nucleotides 2 and 3 of the sense strand;

[0303] b) When the antisense strand is 26 nt in length, counting from the 5' end, nucleotides 1, 2, 5, 9, 17, and 19 of the antisense strand have 2'-F modifications, and the rest have 2'-OMe modifications. There is a phosphorothioate linkage between the 4th and 5th nucleotides. Counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and between the 2nd and 3rd nucleotides.

[0304] In certain embodiments, the RNA inhibitor, wherein the 5'-end of the antisense strand comprises a phosphate or a phosphate mimetic.

[0305] In certain embodiments, the RNA inhibitor, wherein the phosphate mimetic comprises 5'-(E)-vinylphosphonate, 5'-methylphosphonate, (S)-5'-C-methyl analogs and 5'-phosphorothioate (5'-PS).

[0306] In certain embodiments, the RNA inhibitor, wherein the 5'-end of the antisense strand comprises (M06):

[0307] In certain embodiments, the RNA inhibitor, wherein (M06) is linked to the 5'-end of the antisense strand via a phosphate or phosphorothioate bond.

[0308] In certain embodiments, the RNA inhibitor has an invAB modification at the 5'-end of the first nucleotide of the antisense strand.

[0309] In certain embodiments, in the RNA inhibitor, the inverted abasic nucleotide is linked to the 5' end of the first nucleotide of the antisense strand via a phosphorothioate.

[0310] The modified sequences are shown in Table 2.

[0311] Table 2 Modified duplexes

[0312] Among them, the lowercase letters "g," "c," "a," and "u" represent 2'-methoxy-modified nucleotides; the uppercase letters "Gf," "Cf," "Af," and "Uf" represent 2'-fluoro-modified nucleotides; * indicates that the two adjacent nucleotides to the left and right of * are linked by phosphorothioate groups. (D02)*, (InvAB)*, (M06)*, *(D02), *(InvAB), and *(M06) represent D02, invAB, and M06 linked to the nucleotide via phosphorothioate. invAB refers to an inverted abasic nucleotide bonded to the 5' or 3' end of the nucleotide. (M06) refers to a structure described below bonded to the 5' or 3' end of a monomer (e.g., nucleotide):

[0313] (M06) It can be linked to the nucleotide through phosphate or phosphorothioate.

[0314] [D02] refers to a structure bonded to the 5' or 3' end as follows:

[0315] It can be linked to the nucleotide via phosphate or phosphorothioate. Taking 5'-[D02]*A-3' as an example, in some embodiments, its structure is:

[0316] Ligand-coupled RNA inhibitors

[0317] Another aspect of the RNA inhibitors of the present application relates to ways of coupling interfering nucleic acids to ligands to enhance the stability, activity, cellular distribution or cellular uptake of the RNAi agent.

[0318] In certain embodiments, the distribution, targeting, or stability of an RNA inhibitor is altered by introducing a ligand for a target tissue receptor. For example, a specific ligand can provide enhanced affinity for a selected target (e.g., a molecule, cell or cell type, compartment (e.g., a cell or organ compartment, body tissue, organ, or region)) compared to a species in which the ligand is not present.

[0319] The ligand can include naturally occurring substances such as proteins (e.g., human serum albumin (HSA), low-density lipoprotein (LDL) or globulins); carbohydrates (e.g., dextran, pullulan, chitin, chitosan, inulin, cyclodextrin, N-acetylglucosamine, N-acetylgalactosamine or hyaluronic acid); or lipids. The ligand can also be a recombinant or synthetic molecule, such as a synthetic polymer, for example, a synthetic polyamino acid.

[0320] The ligand can also include a targeting group, such as a cell or tissue targeting agent that binds to a specified cell type such as a kidney cell, such as a lectin, glycoprotein, lipid or protein, such as an antibody. The targeting group can be thyrotropin, melanocyte stimulating hormone, lectin, glycoprotein, surfactant protein A, mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine multivalent mannose, multivalent fucose, glycosylated polyamino acids, multivalent galactose, transferrin, bisphosphate, polyglutamic acid, polyaspartic acid, lipid, cholesterol, steroid, bile acid, folic acid, vitamin B12, vitamin A, biotin or RGD peptide or RGD peptide mimetic. In certain embodiments, the ligand is a multivalent galactose, such as N-acetyl-galactosamine.

[0321] The sense strand and antisense strand comprised by the application's RNA inhibitor can be conveniently and routinely prepared by the well-known technology of solid phase synthesis. Any other method known in the art for this type of synthesis, such as liquid phase synthesis or fermentation, can be used additionally or alternatively. It is also known to use similar technology to prepare other oligonucleotides (such as phosphorothioate and alkylated derivatives).

[0322] In certain embodiments, in addition to standard nucleoside phosphoramidite monomers and non-standard nucleoside phosphoramidite monomers that are commercially available and routinely used in oligonucleotide synthesis, the oligonucleotides or linked nucleotides of the present application can be synthesized by an automated synthesizer using a phosphoramidite method derived from ligand-nucleoside phosphoramidite monomers.

[0323] In certain embodiments, the ligand conjugation of the present invention is achieved by coupling the ligand structure to the 5' end and / or 3' end of the antisense strand, and / or the 5' end and / or 3' end of the sense strand.

[0324] For example, the ligand structure can be coupled to the 5' end and / or the 3' end of the sense strand; or the ligand structure can be coupled to the 5' end of the antisense strand and the ligand structure is coupled to the 3' end of the sense strand; or the ligand structure can be coupled to the 3' end of the antisense strand and the ligand is coupled to the 5' end of the sense strand; or the ligand structure is coupled to the 5' end and 3' end of the sense strand; or the ligand structure is coupled to the 3' end of the sense strand.

[0325] In certain embodiments, the ligand described herein is [L96], as shown in the following structural formula:

[0326] In certain embodiments, the ligand described herein is [D02], as shown in the following structural formula:

[0327] In the present invention, [D02] represents the residue of D02 monomer. In certain embodiments, the D02 monomer structure is

[0328] In certain embodiments, [D02] is obtained by Bonded to a nucleotide.

[0329] Taking 5'-[D02]*A-3' as an example, in certain embodiments, its structure is:

[0330] Pharmaceutical composition

[0331] The present application also includes a pharmaceutical composition comprising the RNA inhibitor of the present application or a pharmaceutically acceptable salt thereof.

[0332] In one embodiment, provided herein are pharmaceutical compositions comprising an RNA inhibitor as described herein and a pharmaceutically acceptable excipient.

[0333] Pharmaceutical compositions comprising RNA inhibitors can be used to prevent and / or treat IGF-1R-related disorders, e.g., thyroid eye disease, osteoarthritis, neuropathic pain. Such pharmaceutical compositions are formulated according to the mode of delivery. One example embodiment is a composition formulated for systemic administration by parenteral delivery, e.g., subcutaneous (SC), intramuscular (IM), or intravenous (IV) delivery. The pharmaceutical compositions of the present application can be administered in a dosage sufficient to inhibit IGF-1R gene expression.

[0334] A pharmaceutically acceptable "excipient" or "vegetarian" is a pharmaceutically acceptable solvent, suspending agent, or any other pharmaceutically inert vehicle used to deliver one or more nucleic acids to an animal. Excipients can be liquid or solid and are selected based on the intended mode of administration to provide the desired volume, consistency, etc. when combined with the nucleic acid and other components of a given pharmaceutical composition. RNA inhibitors can be delivered in a manner that targets specific tissues (e.g., hepatocytes).

[0335] In certain embodiments, the pharmaceutical composition further comprises a delivery vehicle (eg, nanoparticles, dendrimers, polymers, liposomes, or cationic delivery systems).

[0336] In certain embodiments, the delivery vehicle comprises a liposome.

[0337] In certain embodiments, the delivery vehicle comprises nanolipids that are capable of forming liposome-nucleic acid nanoparticles with nucleic acid molecules.

[0338] use

[0339] On the other hand, the present application provides the use of the aforementioned RNA inhibitor for inhibiting IGF-1R gene expression or a pharmaceutically acceptable salt thereof and the aforementioned pharmaceutical composition in preparing a drug for preventing or treating a disease or pathology or reducing the risk of a disease or pathology.

[0340] In certain embodiments, the disease or pathology comprises a disease or pathology associated with normal or elevated levels of IGF-1R.

[0341] In certain embodiments, the disease or pathology comprises thyroid eye disease, osteoarthritis, or neuropathic pain.

[0342] In another aspect, the present application provides a method for preventing or treating a disease, condition or syndrome, comprising administering to a subject in need thereof an effective amount of the aforementioned RNA inhibitor for inhibiting IGF-1R gene expression, a pharmaceutically acceptable salt thereof or the aforementioned pharmaceutical composition.

[0343] In certain embodiments, the RNA inhibitor that inhibits IGF-1R gene expression, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is administered to the subject by intraorbital injection, intraarticular injection, intrathecal injection, subcutaneous administration, intravenous administration, oral administration, rectal administration, or intraperitoneal administration.

[0344] In certain embodiments, the method comprises administering to a local or focal area of ​​tissue in a subject in need thereof.

[0345] In certain embodiments, the local or lesion area tissue of the subject in need thereof includes eye tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

[0346] In certain embodiments, the eye tissue is the optic nerve, trabecular meshwork, proximal canal tissue, ganglion, episcleral vein, Schlemm's canal or peripheral eye tissue. Preferably, the joint tissue comprises cartilage tissue, joint connective tissue, and bone tissue; preferably, the central nervous tissue comprises spinal cord tissue and brain tissue; preferably, the peripheral nervous tissue comprises intra-articular nerve tissue and muscle nerve tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

[0347] In certain embodiments, the ocular tissue is retinal ganglion, endothelial cells, periocular muscle, or periocular fat.

[0348] On the other hand, the present application provides a method for inhibiting IGF-1R mRNA or gene expression in cells, tissues or subjects, comprising administering to a subject in need thereof an effective amount of the aforementioned RNA inhibitor for inhibiting IGF-1R gene expression, a pharmaceutically acceptable salt thereof or the aforementioned pharmaceutical composition.

[0349] Cells suitable for treatment using the methods of the present application can be any cell that genetically expresses an IGF-1R gene. Cells suitable for use in the methods of the present application can be mammalian cells, and when contacted with cells that genetically express an IGF-1R gene, the RNAi agent inhibits gene expression of an IGF-1R gene (e.g., a human, primate, non-primate, or rat IGF-1R gene) by at least about 50%, as determined, for example, by PCR or branched DNA (bDNA)-based methods, or by protein-based methods such as immunofluorescence analysis, Western blotting, or flow cytometry.

[0350] As used herein, the term "inhibit" is used interchangeably with "reduce," "reduced," "silenced," "down-regulated," "suppressed," and other similar terms, and encompasses any level of inhibition. Expression of the IGF-1R gene can be assessed based on the level or change in level of any variable associated with IGF-1R gene expression, for example, IGF-1R mRNA levels or IGF-1R protein levels. Such levels can be analyzed in a single cell or in a population of cells (including, for example, a sample derived from a subject). Inhibition can be assessed by a decrease in the absolute or relative level of one or more variables associated with IGF-1R gene expression compared to a control level. The control level can be any type of control level used in the art, for example, a pre-dose baseline level or a level measured from a similar subject, cell, or sample that has not been treated or that has been treated with a control, such as, for example, a buffer-only control or a no active agent control.

[0351] Inhibition of IGF-1R gene expression can be demonstrated by a reduction in the amount of mRNA expressed by a first cell or cell population (such cells may, for example, be present in a sample derived from a subject) in which the IGF-1R gene is transcribed and has been treated (e.g., by contacting one or more cells with an RNA inhibitor of the present invention, or by administering an RNA inhibitor of the present invention to a subject in which such cells are present) such that IGF-1R gene expression is inhibited, compared to a second cell or cell population that is substantially identical to the first cell or cell population but has not been treated in this manner (control cells that have not been treated with the RNA inhibitor or with an RNA inhibitor targeting the gene of interest). In a preferred embodiment, inhibition is assessed in a cell line that highly expresses IGF-1R using an appropriate concentration of siRNA as provided in the Examples, and the mRNA level in the treated cells is expressed as a percentage of the mRNA level in the non-treated control cells.

[0352] In other embodiments, inhibition of IGF-1R gene expression can be assessed by a decrease in a parameter functionally associated with IGF-1R gene expression, e.g., IGF-1R protein levels in the blood or serum of a subject. IGF-1R gene silencing can be measured in any cell that genetically expresses IGF-1R (either endogenous or exogenous from a gene expression construct) and by any assay known in the art.

[0353] Inhibition of IGF-1R protein expression can be demonstrated by a decrease in the level of IGF-1R protein expressed by a cell or cell population or a sample from a subject (e.g., the level of protein in a blood sample from a subject). As described above for assessment of mRNA inhibition, inhibition of protein expression levels in treated cells or cell populations can be similarly expressed as a percentage of the protein level in a control cell or cell population, or as a change in protein level in a sample from a subject (e.g., blood or serum derived therefrom).

[0354] Control cells, cell populations, or subject samples that can be used to evaluate IGF-1R gene inhibition include cells, cell populations, or subject samples that have not been contacted with the RNAi agent of the present application. For example, control cells, cell populations, or subject samples can be derived from a single subject (e.g., a human or animal subject) or an appropriately matched population control prior to treatment with the RNAi agent.

[0355] The level of IGF-1R expressed by a cell or cell population can be measured using any method known in the art for evaluating mRNA gene expression. For example, qRT-PCR can be used to evaluate a decrease in gene expression. A decrease in protein production can be evaluated by any method known in the art, such as ELISA. In certain embodiments, a liver biopsy sample is used as the tissue material for monitoring a decrease in IGF-1R gene or protein gene expression. In other embodiments, a blood sample is used as the subject sample for monitoring a decrease in IGF-1R protein expression.

[0356] Without intending to be bound by any theory, the following examples are merely intended to illustrate the fusion protein, preparation method, and use of the present application, and are not intended to limit the scope of the present invention.

[0357] This application provides the following implementation methods:

[0358] 1. An RNA inhibitor for inhibiting insulin-like growth factor 1 receptor (IGF-1R) gene expression, comprising an antisense strand, wherein the antisense strand forms a complementary region with at least 15 consecutive nucleotides in an mRNA encoding IGF-1R (SEQ ID NO: 828), wherein the complementary region has 0, 1, 2, 3, 4, or 5 mismatches, and preferably the complementary region is 15-30 nucleotide pairs in length, more preferably 17-23 nucleotide pairs in length.

[0359] 2. The RNA inhibitor according to embodiment 1, wherein the antisense strand forms a complementary region with 15, 16, 17, 18, 19, 20, 21, 22 or 23 consecutive nucleotides starting from the 5' end of any one of the following positions in the mRNA encoding IGF-1R (SEQ ID NO: 828):

[0360] Nucleotide position 1118, nucleotide position 1120, nucleotide position 1154, nucleotide position 1367, nucleotide position 1407, nucleotide position 1415, nucleotide position 1532, nucleotide position 1625, nucleotide position 1627, nucleotide position 1628, nucleotide position 1631, nucleotide position 3356, nucleotide position 3357, nucleotide position 3359, nucleotide position 3792, nucleotide position 4198, nucleotide position 4200, nucleotide position 4208, nucleotide position 4685, nucleotide position 5246, nucleotide position 5392, nucleotide position 5393, nucleotide position 6329, nucleotide position 6332, nucleotide position 6333 , nucleotide No. 6334, nucleotide No. 10215, nucleotide No. 10738, nucleotide No. 10740, nucleotide No. 10770, nucleotide No. 10772, nucleotide No. 10773, nucleotide No. 10946, nucleotide No. 10956, nucleotide No. 10957, nucleotide No. 1418, nucleotide No. 1541, nucleotide No. 2050, nucleotide No. 2055, nucleotide No. 2056, nucleotide No. 2233, nucleotide No. 2234, nucleotide No. 2456, nucleotide No. 2591, nucleotide No. 2603, nucleotide No. 2606, nucleotide No. 2609, nucleotide No. 2776, nucleotide No. 2890, Nucleotide No. 2906, Nucleotide No. 2909, Nucleotide No. 2957, Nucleotide No. 2960, Nucleotide No. 3008, Nucleotide No. 3195, Nucleotide No. 3398, Nucleotide No. 3608, Nucleotide No. 3609, Nucleotide No. 3650, Nucleotide No. 3660, Nucleotide No. 3815, Nucleotide No. 3882, Nucleotide No. 3996, Nucleotide No. 4067, Nucleotide No. 4121, Nucleotide No. 4122, Nucleotide No. 4292, Nucleotide No. 4293, Nucleotide No. 4295, Nucleotide No. 4516, Nucleotide No. 4519, Nucleotide No. 5163, Nucleotide No. 5309, Nucleotide No. 5310, Nucleotide No. 5377, Nucleotide No. 5380, Nucleotide No. 5381, Nucleotide No. 5663, Nucleotide No. 5700, Nucleotide No. 5711, Nucleotide No. 5712, Nucleotide No. 5721, Nucleotide No. 5896, Nucleotide No. 5898, Nucleotide No. 6203, Nucleotide No. 6236, Nucleotide No. 6237, Nucleotide No. 6240, Nucleotide No. 6245, Nucleotide No. 6288, Nucleotide No. 6299, Nucleotide No. 6305, Nucleotide No. 6309, Nucleotide No. 6310, Nucleotide No. 6343, Nucleotide No. 6363, Nucleotide No. 6365, Nucleotide No. 6366, Nucleotide No. 6373,Nucleotide No. 6379, Nucleotide No. 6381, Nucleotide No. 6431, Nucleotide No. 6434, Nucleotide No. 6496, Nucleotide No. 6518, Nucleotide No. 6755, Nucleotide No. 6760, Nucleotide No. 6883, Nucleotide No. 6885, Nucleotide No. 6947, Nucleotide No. 6948, Nucleotide No. 6949, Nucleotide No. 6952, Nucleotide No. 6953, Nucleotide No. 6954, Nucleotide No. 7574, Nucleotide No. 7621, Nucleotide No. 7764, Nucleotide No. 7830, Nucleotide No. 7906, Nucleotide No. 8540, Nucleotide No. 8836, Nucleotide No. 8892, Nucleoside No. 8975 Acid, nucleotide No. 8977, nucleotide No. 9163, nucleotide No. 9175, nucleotide No. 9186, nucleotide No. 9265, nucleotide No. 9267, nucleotide No. 9340, nucleotide No. 9347, nucleotide No. 9352, nucleotide No. 9356, nucleotide No. 9478, nucleotide No. 9480, nucleotide No. 9606, nucleotide No. 9611, nucleotide No. 9634, nucleotide No. 9680, nucleotide No. 9681, nucleotide No. 9724, nucleotide No. 9725, nucleotide No. 9814, nucleotide No. 10015, nucleotide No. 10016, nucleotide No. 10189, nucleotide No. 10244, nucleotide No. 10 nucleotide 10526, nucleotide 10528, nucleotide 10534, nucleotide 10535, nucleotide 10537, nucleotide 10540, nucleotide 10541, nucleotide 10679, nucleotide 10694, nucleotide 10717, nucleotide 10728, nucleotide 10729, nucleotide 10730, nucleotide 10754, nucleotide 10763, nucleotide 10766, nucleotide 10942, nucleotide 11034, nucleotide 11078, nucleotide 11126, nucleotide 11156 , nucleotide 11159, nucleotide 11160, nucleotide 11292, nucleotide 11340, nucleotide 11344, nucleotide 11383, nucleotide 11395, nucleotide 11789, nucleotide 11864, nucleotide 11865, nucleotide 11939, nucleotide 12092, nucleotide 12116, nucleotide 12130, nucleotide 12154, nucleotide 12156, nucleotide 12177, nucleotide 12195, nucleotide 1164, nucleotide 1163, nucleotide 1162, nucleotide 1161, nucleotide 1160,Nucleotide 1165, nucleotide 1166, nucleotide 1167, nucleotide 1168.

[0361] 3. The RNA inhibitor according to any one of embodiments 1-2, wherein the antisense strand forms a complementary region with the sequence between nucleotides 1160 and 1188, between nucleotides 6235 and 6465, or between nucleotides 6329 and 6455 of the mRNA encoding IGF-1R (SEQ ID NO: 828) counting from the 5' end.

[0362] 4. The RNA inhibitor according to any one of embodiments 1 to 3, wherein the RNA inhibitor is ribonucleic acid, and further, the RNA inhibitor is single-stranded ribonucleic acid or double-stranded ribonucleic acid.

[0363] 5. The RNA inhibitor according to any one of embodiments 1 to 4, wherein the RNA inhibitor is an antisense oligonucleotide (ASO), shRNA, miRNA or siRNA.

[0364] 6. An RNA inhibitor according to any one of embodiments 1 to 5, comprising a sense strand capable of forming a complementary duplex with the antisense strand, wherein the antisense strand comprises a sequence that forms a duplex complementary region with at least 15 consecutive nucleotides in the sense strand sequence, and the duplex complementary region has 0, 1, 2, 3, 4 or 5 mismatches. Preferably, the duplex complementary region is 15-30 nucleotide pairs in length, more preferably 17-23 nucleotide pairs in length.

[0365] 7. The RNA inhibitor according to embodiment 6, wherein the sense strand and the antisense strand are present on two different nucleic acid strands, preferably the RNA inhibitor is siRNA or shRNA, more preferably the RNA inhibitor is siRNA.

[0366] 8. The RNA inhibitor according to any one of embodiments 6-7, wherein the sense strand and the antisense strand are present on the same nucleic acid strand, and preferably the RNA inhibitor is shRNA.

[0367] 9. The RNA inhibitor according to any one of embodiments 6-8, wherein the total length of the sense strand is 15-50 nucleotides, preferably the total length of the sense strand is 16-30 nucleotides, and more preferably the total length of the sense strand is 17, 18, 19, 20 or 21 nucleotides.

[0368] 10. The RNA inhibitor according to any one of embodiments 6-9, wherein the total length of the antisense strand is 19-50 nucleotides, preferably the total length of the antisense strand is 19-30 nucleotides, and more preferably the total length of the antisense strand is 21, 22, 23, 24, 25, 26 or 27 nucleotides.

[0369] 11. The RNA inhibitor according to any one of embodiments 6 to 10, wherein the sense strand and antisense strand each independently optionally comprise a 3' or 5' overhang of 1, 2 or 3 nucleotides.

[0370] 12. The RNA inhibitor of embodiment 11, wherein both the sense strand and the antisense strand have a 3' overhang of 1-3 nucleotides in length, or the sense strand has a 3' or 5' overhang of 1-3 nucleotides in length, or the antisense strand has a 3' or 5' overhang of 1-3 nucleotides in length.

[0371] 13. The RNA of any one of embodiments 1-12, wherein the antisense strand comprises at least 15 consecutive nucleotides of the sequence of any one of SEQ ID NOs: 199-400.

[0372] 14. The RNA inhibitor according to any one of embodiments 6-13, wherein the sense strand comprises at least 15 consecutive nucleotides of the sequence of any one of SEQ ID NOs: 1-198.

[0373] 15. The RNA inhibitor according to any one of embodiments 1 to 14, comprising a duplex selected from the group consisting of: ds-n1, ds-n2, ds-n3, ds-n4, ds-n5, ds-n6, ds-n7, ds-n8, ds-n9, ds-n10, ds-n11, ds-n12, ds-n13, ds-n14, ds-n15, ds-n16, ds-n17, ds-n18, ds-n19, ds-n20, ds-n21, ds-n22, ds-n23, ds-n24, ds-n25, ds-n26, ds-n27, ds-n28, ds-n29, ds-n30, ds-n31, ds-n32, ds-n33, ds-n34, ds-n35, ds-n36, ds-n37, ds-n38, ds-n39, ds-n40 , ds-n41, ds-n42, ds-n43, ds-n44, ds-n45, ds-n46, ds-n47, ds-n48, ds-n4 9. ds-n50, ds-n51, ds-n52, ds-n53, ds-n54, ds-n55, ds-n56, ds-n57, ds-n 58. ds-n59, ds-n60, ds-n61, ds-n62, ds-n63, ds-n64, ds-n65, ds-n66, ds-n 67. ds-n68, ds-n69, ds-n70, ds-n71, ds-n72, ds-n73, ds-n74, ds-n75, ds- n76, ds-n77, ds-n78, ds-n79, ds-n80, ds-n81, ds-n82, ds-n83, ds-n84, ds -n85, ds-n86, ds-n87, ds-n88, ds-n89, ds-n90, ds-n91, ds-n92, ds-n93, d s-n94, ds-n95, ds-n96, ds-n97, ds-n98, ds-n99, ds-n100, ds-n101, ds-n10 2. ds-n103, ds-n104, ds-n105, ds-n106, ds-n107, ds-n108, ds-n109, ds-n 110, ds-n111, ds-n112, ds-n113, ds-n114, ds-n115, ds-n116, ds-n117, ds- n118, ds-n119, ds-n120, ds-n121, ds-n122, ds-n123, ds-n124, ds-n125, d s-n126, ds-n127, ds-n128, ds-n129, ds-n130, ds-n131, ds-n132, ds-n133,ds-n134, ds-n135, ds-n136, ds-n137, ds-n138, ds-n139, ds-n140, ds-n141, ds-n1 42. ds-n143, ds-n144, ds-n145, ds-n146, ds-n147, ds-n148, ds-n149, ds-n150, ds -n151, ds-n152, ds-n153, ds-n154, ds-n155, ds-n156, ds-n157, ds-n158, ds-n159 , ds-n160, ds-n161, ds-n162, ds-n163, ds-n164, ds-n165, ds-n166, ds-n167, ds-n 168, ds-n169, ds-n170, ds-n171, ds-n172, ds-n173, ds-n174, ds-n175, ds-n176, d s-n177, ds-n178, ds-n179, ds-n180, ds-n181, ds-n182, ds-n183, ds-n184, ds-n18 5. ds-n186, ds-n187, ds-n188, ds-n189, ds-n190, ds-n191, ds-n192, ds-n193, ds- n194, ds-n195, ds-n196, ds-n197, ds-n198, ds-n201, ds-n202, ds-n203, ds-n204. ,

[0374] 16. The RNA inhibitor of embodiment 15, comprising a duplex selected from the group consisting of ds-n17, ds-n23, ds-n31, ds-n39, ds-n45, ds-n58, ds-n60, ds-n64, ds-n67, ds-n68, ds-n73, ds-n74, ds-n76, ds-n77, ds-n80, ds-n81, ds-n83, ds-n84, ds-n86, ds-n88, ds-n89, ds-n 98. ds-n99, ds-n103, ds-n104, ds-n112, ds-n113, ds-n115, ds-n153, ds-n154, ds-n156, ds-n157, ds-n163, ds- n164, ds-n165, ds-n188, ds-n190, ds-n191, ds-n192, ds-n193, ds-n194, ds-n195, ds-n196, ds-n197, ds-n198.

[0375] 17. The RNA inhibitor of embodiment 15, comprising a duplex selected from the group consisting of ds-n17, ds-n23, ds-n31, ds-n39, ds-n45, ds-n58, ds-n60, ds-n64, ds-n67, ds-n68, ds-n73, ds-n74, ds-n76, ds-n77, ds-n80, ds-n81, ds-n82, ds-n83, ds-n84, ds-n85, ds-n86, ds-n87, ds-n88, ds-n90, ds-n91, ds-n92, ds-n93, ds-n94, ds-n96, ds-n97, ds-n98 n83, ds-n84, ds-n86, ds-n88, ds-n89, ds-n98, ds-n99, ds-n103, ds-n104, ds-n112, ds-n1 13. ds-n115, ds-n153, ds-n154, ds-n156, ds-n157, ds-n163, ds-n164, ds-n165, ds-n188.

[0376] 18. The RNA inhibitor of embodiment 16, comprising a duplex selected from the group consisting of ds-n23, ds-n58, ds-n67, ds-n86, ds-n89, ds-n98, ds-n99, ds-n103, ds-n190, ds-n198, and ds-n113.

[0377] 19. The RNA inhibitor according to any one of embodiments 1-18, characterized in that the antisense strand further includes region B1 at the 3' end, and the sense strand further includes region A1 at the 5' end, wherein region A1 is 0-6 nucleotides and region B1 is 0-6 nucleotides.

[0378] 20. The RNA inhibitor according to embodiment 19, characterized in that the region B1 is 0, 1, 2, 3, 4, 5 or 6 nucleotides; preferably, the region B1 is 0 or 2 nucleotides.

[0379] 21. The RNA inhibitor according to any one of embodiments 16-20, characterized in that the region A1 is 0, 1, 2, 3, 4, 5 or 6 nucleotides; preferably, the region A1 is 0 or 2 nucleotides.

[0380] 22. The RNA inhibitor according to embodiment 21, characterized in that the antisense strand further includes region X2 at the 5' end, and the sense strand or sense nucleic acid further includes region X1 at the 3' end, and region X1 and region X2 are complementary to each other.

[0381] 23. The RNA inhibitor of embodiment 22, wherein X1 is A, U, modified A, or modified U, and X2 is U, A, modified U, or modified A.

[0382] 24. The RNA inhibitor according to any one of embodiments 19-23, characterized in that the antisense strand comprises X2, Y, Z and N in sequence from 3' to 5' direction at the 5' end, wherein X2 and Y are independently A, U, modified A or modified U, Z is G or modified G, and N comprises at least one nucleotide.

[0383] 25. The RNA inhibitor according to any one of embodiments 19-24, wherein X2, Y, and Z are AAG, AUG, UUG, or UAG, or partially or fully modified versions of the above sequences, in the 3' to 5' direction.

[0384] 26. The RNA inhibitor according to any one of embodiments 21-25, wherein X2, Y, and Z are AAG, AUG, UUG, or UAG, or partially or fully modified versions of the above sequences, in the 3' to 5' direction.

[0385] 27. The RNA inhibitor according to any one of embodiments 21-26, wherein N is C or modified C.

[0386] 28. The RNA inhibitor according to any one of embodiments 21-27, wherein X2, Y, Z and N are AAGC and UAGC in the 3' to 5' direction, or partially or fully modified versions of the above sequences.

[0387] 29. The RNA inhibitor according to any one of embodiments 16-28, wherein the antisense strand comprises at least 15 consecutive nucleotides of the sequence as described in any one of SEQ ID NOs: 401-413.

[0388] 30. The RNA inhibitor of any one of embodiments 1-29, which is selected from ds-n190-1, ds-n191-1, ds-n192-1, ds-n193-1, ds-n194-1, ds-n195-1, ds-n196-1, ds-n197-1, ds-n198-1, ds-n201-1, ds-n202-1, ds-n203-1, and ds-n204-1.

[0389] 31. The RNA inhibitor according to any one of embodiments 1-30, wherein at least one nucleotide in the RNA inhibitor is a modified nucleotide.

[0390] 32. The RNA inhibitor according to any one of embodiments 1-31, wherein at least 70%, 80%, 90%, 95% of the nucleotides in the RNA inhibitor are modified nucleotides; preferably all nucleotides are modified nucleotides.

[0391] 33. An RNA inhibitor according to any one of embodiments 31-32, wherein the modification comprises a combination of one or more of the following: 2'-OMe (2'-O-methyl) modification, 2'-F (2'-deoxy-2'-fluoro) modification, 2'-O-MOE (2'-O-methoxyethyl) modification, 2'-deoxy (2'-d) modification, 5'-morpholine (5'-Mo) modification, unlocked nucleic acid (UNA) modification, glycol nucleic acid (GNA) modification, locked nucleic acid (LNA) modification, tricyclic DNA (tcDNA) modification, (S)-constrained ethyl bicyclic nucleic acid ((S)-cEt-BNA) modification, phosphorothioate (PS) modification, phosphorodithioate (PS2) modification, methylphosphonate (MP) modification, methoxypropylmethylphosphonate (MOP) modification, peptide nucleic acid (PNA) modification, 5'-(E)-vinyl phosphate (VP) modification. ) modification (VP), N6-methyladenosine (m6A) modification, 5-methylcytidine (m5C) modification, 3-methyluridine (m3U) modification, 5-methyluridine (m5U) modification, pseudouridine modification, 2-thiouridine (s2U) modification, propyne uridine (5-pU) modification, inverted abasic nucleotide (invAB) modification by bonding the 5' or 3' end of the nucleotide, replacing the nucleotide with an inverted abasic nucleotide (invAb) modification, replacing the nucleotide with a 2,4-difluoromethylphenyl ribonucleotide (rF) modification or replacing the nucleotide with a (S)-glycerol nucleic acid modification, preferably a 2'-OMe modification, a 2'-F modification, a 2'-deoxy modification, a VP modification, a 5'-MP modification, a PS modification, a PS2 modification, an MP modification, a MOP modification, an M06 modification, an invAb modification or an invAB modification.

[0392] 34. The RNA inhibitor of any one of embodiments 31-33, wherein the 3' or 5'-end of the antisense strand comprises a phosphate or a phosphate mimetic, or the 3' or 5'-end of the sense strand comprises a phosphate or a phosphate mimetic.

[0393] 35. The RNA inhibitor of embodiment 34, wherein the phosphate mimetic comprises 5'-(E)-vinylphosphonate, 5'-methylphosphonate, (S)-5'-C-methyl analogs, and 5'-phosphorothioate (5'-PS).

[0394] 36. The RNA inhibitor according to any one of embodiments 31-35, wherein the 5'-end of the antisense strand comprises (M06) modification:

[0395] 37. The RNA inhibitor according to any one of embodiments 31-36, wherein an invAB modification is present at the 3' or 5'-end of the antisense strand, or an invAB modification is present at the 3' or 5'-end of the sense strand.

[0396] 38. The RNA inhibitor according to any one of embodiments 31 to 37, wherein the inverted abasic nucleotide or M06 is linked to the 3' or 5' end of the antisense strand or the 3' or 5' end of the sense strand via a phosphorothioate.

[0397] 39. The RNA inhibitor according to any one of embodiments 31-38, having the following modification combination

[0398] a) the sense strand comprises a first sequence of 16-21 nt in length, wherein the first sequence comprises the following modifications: 2'-F modifications are present at nucleotides 9, 10, and 11, counting from the 5' end;

[0399] b) the antisense strand comprises a second sequence of 16-21 nt in length, wherein the second sequence includes the following modifications: counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, a phosphorothioate linkage between nucleotides 2 and 3, and a phosphorothioate linkage between nucleotides 3 and 4; and counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2; and counting from the 5' end, there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16.

[0400] 40. The RNA inhibitor according to embodiment 39, wherein the length of the first sequence is preferably 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, preferably 21 nt.

[0401] 41. The RNA inhibitor according to embodiment 39, wherein the second sequence length is preferably 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, preferably 21 nt.

[0402] 42. The RNA inhibitor according to any one of embodiments 39-41, having the following modification combination

[0403] a) The sense strand is 21 nt long and includes the following modifications: 2'-F modifications are present at nucleotides 9, 10, and 11, counting from the 5' end;

[0404] b) The antisense strand is 21 nt in length and includes the following modifications: counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, a phosphorothioate linkage between nucleotides 2 and 3, and a phosphorothioate linkage between nucleotides 3 and 4; and, counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2; and, counting from the 5' end, there is a 2'-F modification at nucleotides 2, 3, 4, 12, 14, and 16.

[0405] 43. The RNA inhibitor according to any one of embodiments 28-42, wherein the antisense strand comprises at least 15 consecutive nucleotides of the sequence as described in any one of SEQ ID NOs: 624-812.

[0406] 44. The RNA inhibitor according to any one of embodiments 28-42, wherein the sense strand comprises at least 15 consecutive nucleotides of the sequence as described in any one of SEQ ID NOs: 414-602.

[0407] 45. The RNA inhibitor of any one of embodiments 28-44, which is selected from the group consisting of: ds-m1, ds-m2, ds-m3, ds-m4, ds-m5, ds-m6, ds-m7, ds-m8, ds-m9, ds-m10, ds-m11, ds-m12, ds-m13, ds-m14, ds-m15, ds-m16, ds-m17, ds-m18, ds-m19, ds-m20, ds-m21, ds-m22, ds-m23, ds-m24, ds-m25, ds-m26, ds-m27, ds-m28, ds-m29, ds-m30, ds-m31, ds-m32, ds-m33, -m33, ds-m34, ds-m35, ds-m36, ds-m37, ds-m38, ds-m39, ds-m40, ds-m41, d s-m42, ds-m43, ds-m44, ds-m45, ds-m46, ds-m47, ds-m48, ds-m49, ds-m50, ds-m51, ds-m52, ds-m53, ds-m54, ds-m55, ds-m56, ds-m57, ds-m58, ds-m59 , ds-m60, ds-m61, ds-m62, ds-m63, ds-m64, ds-m65, ds-m66, ds-m67, ds-m68 , ds-m69, ds-m70, ds-m71, ds-m72, ds-m73, ds-m74, ds-m75, ds-m76, ds-m7 7. ds-m78, ds-m79, ds-m80, ds-m81, ds-m82, ds-m83, ds-m84, ds-m85, ds-m 86. ds-m87, ds-m88, ds-m89, ds-m90, ds-m91, ds-m92, ds-m93, ds-m94, ds- m95, ds-m96, ds-m97, ds-m98, ds-m99, ds-m100, ds-m101, ds-m102, ds-m103 , ds-m104, ds-m105, ds-m106, ds-m107, ds-m108, ds-m109, ds-m110, ds-m1 11. ds-m112, ds-m113, ds-m114, ds-m115, ds-m116, ds-m117, ds-m118, ds- m119, ds-m120, ds-m121, ds-m122, ds-m123, ds-m124, ds-m125, ds-m126, d s-m127, ds-m128, ds-m129, ds-m130, ds-m131, ds-m132, ds-m133, ds-m134,ds-m135, ds-m136, ds-m137, ds-m138, ds-m139, ds-m140, ds-m141, ds-m142, ds-m143, ds-m144, ds-m145, ds-m146, ds-m147, ds-m14 8. ds-m149, ds-m150, ds-m151, ds-m152, ds-m153, ds-m154, ds-m155, ds-m156, ds-m157, ds-m158, ds-m159, ds-m160, ds-m161, ds-m 162, ds-m163, ds-m164, ds-m165, ds-m166, ds-m167, ds-m168, ds-m169, ds-m170, ds-m171, ds-m172, ds-m173, ds-m174, ds-m175, ds -m176, ds-m177, ds-m178, ds-m179, ds-m180, ds-m181, ds-m182, ds-m183, ds-m184, ds-m185, ds-m186, ds-m187, ds-m188, ds-m189. ,

[0408] 46. ​​The RNA inhibitor according to any one of embodiments 28-39, wherein

[0409] a) the sense strand comprises a third sequence of 18-21 nt in length, wherein the third sequence comprises the following modifications: counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, there is a phosphorothioate linkage between the 2nd and 3rd nucleotides, and there is a phosphorothioate linkage between the 9th, 10th, 11th, and 18th nucleotides of the sense strand.

[0410] 2'-F modification, the rest are 2'-OMe modification; counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and a phosphorothioate linkage between the 2nd and 3rd nucleotides;

[0411] b) the antisense strand comprises a fourth sequence of 19-26 nt in length, wherein the fourth sequence includes the following modifications: counting from the 5' end, nucleotides 1, 2, 5, 9, 17, and 19 of the antisense strand are 2'-F modified, the rest are 2'-OMe modified, and there is a phosphorothioate linkage between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides.

[0412] 47. The RNA inhibitor according to embodiment 46, wherein the length of the third sequence is preferably 18 nt, 19 nt, 20 nt, 21 nt, 22 nt, 21 nt, preferably 21 nt.

[0413] 48. The RNA inhibitor according to embodiment 47, wherein the length of the fourth sequence is preferably 19 nt, 20 nt, 21 nt, 22 nt, 23 nt, 24 nt, 25 nt, 26 nt, preferably 26 nt.

[0414] 49. The RNA inhibitor according to any one of embodiments 46-48, wherein

[0415] a) The sense strand is 21 nt long. Counting from the 5' end, there is a phosphorothioate linkage between nucleotides 1 and 2, and between nucleotides 2 and 3. Nucleotides 9, 10, 11, and 18 of the sense strand are 2'-F modified, and the rest are 2'-OMe modified. Counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2, and between nucleotides 2 and 3.

[0416] b) The antisense strand is 26 nt in length. Counting from the 5' end, nucleotides 1, 2, 5, 9, 17, and 19 of the antisense strand have 2'-F modifications, and the rest have 2'-OMe modifications. There is a phosphorothioate linkage between the 4th and 5th nucleotides. Counting from the 3' end, there is a phosphorothioate linkage between nucleotides 1 and 2, and between nucleotides 2 and 3.

[0417] 50. The RNA inhibitor according to any one of embodiments 31-49, wherein the antisense strand comprises at least 15 consecutive nucleotides of the sequence as described in any one of SEQ ID NOs: 813-827.

[0418] 51. The RNA inhibitor according to any one of embodiments 31-50, wherein the sense strand comprises at least 15 consecutive nucleotides of the sequence as described in any one of SEQ ID NOs: 603-617.

[0419] 52. The RNA inhibitor of any one of embodiments 31-51, comprising a duplex selected from the group consisting of ds-m190, ds-m191, ds-m192, ds-m193, ds-m194, ds-m195, ds-m196, ds-m197, ds-m198, ds-m199, ds-m200, ds-m201, ds-m202, ds-m203, and ds-m204.

[0420] 53. The RNA inhibitor according to any one of embodiments 1-52 further comprises a delivery system, wherein the delivery system is conjugated to the sense chain and / or antisense chain, and the delivery system enables the RNA inhibitor to reach the target RNA in the target tissue to produce a gene silencing effect.

[0421] 54. The RNA inhibitor according to embodiment 53, wherein the target tissue is eye tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

[0422] 55. The RNA inhibitor according to embodiment 54, wherein the eye tissue is the optic nerve, trabecular meshwork, proximal canal tissue, ganglion, episcleral vein, Schlemm's canal, or peripheral eye tissue; preferably, the joint tissue comprises cartilage tissue, joint connective tissue, and bone tissue; preferably, the central nervous tissue comprises spinal cord tissue and brain tissue; preferably, the peripheral nervous tissue comprises intra-articular nerve tissue and muscle nerve tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

[0423] 56. The RNA inhibitor of embodiment 55, wherein the eye tissue is retinal ganglion, endothelial cells, periocular muscle, or periocular fat.

[0424] 57. The RNA inhibitor according to any one of embodiments 53-56, wherein the delivery system is independently conjugated to one or more internal positions of the double-stranded ribonucleic acid.

[0425] 58. The RNA inhibitor of embodiment 57, wherein the internal position is on a nucleobase, a sugar ring, a methylphosphonate bond, a phosphorothioate diester bond, or a phosphodiester bond.

[0426] 59. The RNA inhibitor according to any one of embodiments 53-58, wherein the delivery system is a lipophilic structure comprising a lipophilic group and a linker, and the lipophilic group is connected to the double-stranded ribonucleic acid via the linker.

[0427] 60. The RNA inhibitor according to embodiment 59, wherein the lipophilic structure is selected from aliphatic, alicyclic and polyalicyclic compounds.

[0428] 61. The RNA inhibitor according to embodiment 60, wherein the lipophilic structure contains saturated or unsaturated C16 or C22.

[0429] 62. The RNA inhibitor of any one of embodiments 59-61, wherein the linker is selected from a single bond, an ether, a thioether, a urea, a carbonate, an amine, an amide, a maleimide-thioether, a disulfide, a phosphodiester, a sulfonamide bond, a product of a click reaction, and a carbamate.

[0430] 63. The RNA inhibitor according to embodiments 53-62, wherein the delivery system is

[0431] 64. The RNA inhibitor according to any one of embodiments 53-63, wherein the delivery system is conjugated to the 5' end and / or 3' end of the antisense strand or antisense nucleic acid fragment.

[0432] 65. The RNA inhibitor according to any one of embodiments 53-64, wherein the delivery system is conjugated to the 5' end and / or 3' end of the sense strand or sense nucleic acid fragment.

[0433] 66. An RNA inhibitor according to any one of embodiments 53-65, wherein the delivery system is conjugated to the 5' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 3' end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

[0434] 67. An RNA inhibitor according to any one of embodiments 53-66, wherein the delivery system is conjugated to the 3' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 5' end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

[0435] 68. An RNA inhibitor according to any one of embodiments 53-67, wherein the delivery system is conjugated to the 5' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 5' end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

[0436] 69. An RNA inhibitor according to any one of embodiments 53-68, wherein the delivery system is conjugated to the 3' end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 3' end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

[0437] 70. The RNA inhibitor according to any one of embodiments 53-69, wherein the number of delivery systems is 1, 2, 3, 4, 5 or 6.

[0438] 71. The RNA inhibitor of any one of embodiments 53-70, wherein the antisense strand comprises at least 15 consecutive nucleotides of the sequence of any one of SEQ ID NOs: 822-827.

[0439] 72. The RNA inhibitor of any one of embodiments 53-71, wherein the sense strand comprises at least 15 consecutive nucleotides of the sequence of any one of SEQ ID NOs: 618-623.

[0440] 73. The RNA inhibitor according to any one of embodiments 53-72, which is selected from: Z1, Z2, Z3, Z4, Z5, Z6.

[0441] 74. A pharmaceutical composition comprising the RNA inhibitor according to any one of embodiments 1-73, and / or a physiologically acceptable excipient and / or carrier and / or diluent.

[0442] 75. The composition according to embodiment 74 is characterized in that the pharmaceutically acceptable carrier includes or is selected from aqueous carriers, liposomes, high molecular polymers or polypeptides.

[0443] 76. Use of an RNA inhibitor targeting IGF-1R and a pharmaceutical composition thereof in the preparation of a drug for treating IGF-1R-related diseases or pathologies; preferably, the RNA inhibitor is siRNA; more preferably, the siRNA is administered to a local or lesion area tissue of a subject in need.

[0444] 77. The use according to embodiment 76, wherein the IGF-1R-related disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels.

[0445] 78. The use according to any one of embodiments 76-77, wherein the IGF-1R related disease or pathology comprises thyroid eye disease, osteoarthritis, and neuropathic pain.

[0446] 79. The use according to any one of embodiments 76-78, wherein the RNA inhibitor and the pharmaceutical composition thereof are the RNA inhibitor according to any one of embodiments 1-73 or the pharmaceutical composition according to any one of embodiments 74-75.

[0447] 80. Use of the RNA inhibitor of any one of embodiments 1-73 or the pharmaceutical composition of any one of embodiments 74-75 in the preparation of a medicament for preventing or treating a disease or pathology or reducing the risk of a disease or symptom.

[0448] 81. The use according to embodiment 80, wherein the disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels.

[0449] 82. The use according to any one of embodiments 80-81, wherein the disease or pathology comprises thyroid eye disease, osteoarthritis, or neuropathic pain.

[0450] 83. A method for preventing or treating an IGF-1R-related disease or symptom, comprising administering an effective amount of an RNA inhibitor and a pharmaceutical composition thereof to a subject in need thereof.

[0451] 84. The method according to embodiment 83, wherein the RNA inhibitor and the pharmaceutical composition thereof are the RNA inhibitor described in any one of embodiments 1-73 or the pharmaceutical composition described in any one of embodiments 74-75.

[0452] 85. The method of embodiments 83-84, wherein the RNA inhibitor, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is administered to the subject by intraorbital injection, intraarticular injection, intrathecal injection, subcutaneous injection, intravenous injection, oral administration, rectal administration, or intraperitoneal administration.

[0453] 86. The method of any one of embodiments 83-85, comprising administering to a local or focal area of ​​tissue in a subject in need thereof.

[0454] 87. The method of embodiment 86, wherein the local or lesion area tissue of the subject in need includes eye tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

[0455] 88. The method according to embodiment 87, wherein the eye tissue is the optic nerve, trabecular meshwork, proximal canal tissue, ganglion, episcleral vein, Schlemm's canal or peripheral eye tissue; preferably, the joint tissue comprises cartilage tissue, joint connective tissue, and bone tissue; preferably, the central nervous tissue comprises spinal cord tissue and brain tissue; preferably, the peripheral nervous tissue comprises intra-articular nerve tissue and muscle nerve tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

[0456] 89. The method of embodiment 88, wherein the ocular tissue is retinal ganglion, endothelial cells, peripheral ocular muscle, or peripheral ocular fat.

[0457] 90. The method of any one of embodiments 83-89, wherein the IGF-1R-related disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels.

[0458] 91. A method according to any one of embodiments 83-90, wherein the IGF-1R related disease or pathology includes thyroid eye disease, osteoarthritis, and neuropathic pain.

[0459] 92. A method for inhibiting IGF-1R gene expression in a cell, tissue, or subject, comprising administering to the cell, tissue, or subject an effective amount of the RNA inhibitor of any one of embodiments 1-73 or the pharmaceutical composition of any one of embodiments 74-75.

[0460] 93. The method of embodiment 92, wherein the disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels.

[0461] 94. The method of embodiment 93, wherein the disease or pathology comprises thyroid eye disease, osteoarthritis, or neuropathic pain.

[0462] 95. A method for preventing or treating a disease or symptom, the method comprising administering to a subject in need thereof an effective amount of an RNA inhibitor according to any one of embodiments 1-73 or a pharmaceutical composition according to any one of embodiments 74-75.

[0463] 96. The method of embodiment 95, wherein the RNA inhibitor, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is administered to the subject by intraorbital injection, intraarticular injection, intrathecal injection, subcutaneous injection, intravenous injection, oral administration, rectal administration, or intraperitoneal administration.

[0464] 97. The method of any one of embodiments 95-96, comprising administering to a local or focal area of ​​tissue in a subject in need thereof.

[0465] 98. The method of embodiment 97, wherein the local or lesion area tissue of the subject in need includes eye tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

[0466] 99. The method according to embodiment 98, wherein the eye tissue is the optic nerve, trabecular meshwork, proximal canal tissue, ganglion, episcleral vein, Schlemm's canal or peripheral eye tissue; preferably, the joint tissue comprises cartilage tissue, joint connective tissue, and bone tissue; preferably, the central nervous tissue comprises spinal cord tissue and brain tissue; preferably, the peripheral nervous tissue comprises intra-articular nerve tissue and muscle nerve tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

[0467] 100. The method of embodiment 99, wherein the ocular tissue is retinal ganglion, endothelial cells, peripheral ocular muscle, or peripheral ocular fat.

[0468] Example

[0469] Example 1 Design and synthesis of siRNA molecules

[0470] The human transcript of the IGF-1R gene was obtained (from the NCBI website, transcript number: NM_000875.5), and the original sequence of siRNA targeting the IGF-1R gene was designed based on the full-length region of human IGF-1R mRNA (including 5'-UTR, CDS, and 3'-UTR).

[0471] Oligoribonucleotides were synthesized using the phosphoramidite solid phase synthesis technique. Synthesis was performed on universal controlled pore glass (CPG). All 2'-modified RNA phosphoramidites and auxiliary reagents were commercially available. All phosphoramidites were dissolved in anhydrous acetonitrile and molecular sieves were added. The coupling time was 1.0 min using 5-ethylthio-1H-tetrazole as an activator. Phosphorothioate bonds were generated using a 50 mM solution of 3-((dimethylamino-methylene)amino)-3H-1,2,4-dithiazole-3-thione in anhydrous acetonitrile / pyridine (v / v = 1 / 1) with a reaction time of 1.8 min. All sequences were synthesized after the final removal of the DMT group.

[0472] Cleavage and deprotection of oligomers bound to CPG: After solid-phase synthesis is completed, the protecting groups are removed by treatment with acetonitrile containing 20% ​​diethylamine for 30 minutes, without cleaving the oligonucleotide from the CPG. Subsequently, the dried CPG is treated with concentrated aqueous ammonia at 40 degrees Celsius for 18 hours. After centrifugation, the supernatant is transferred to a new tube and the CPG is washed with aqueous ammonia. The combined solution is concentrated to obtain a solid mixture.

[0473] Purification of single-stranded oligoribonucleotides: Oligomers were purified by HPLC using a NanoQ anion exchange column. Buffer A consisted of 10 mM sodium perchlorate, 20 mM Tris, 1 mM EDTA, pH 7.4, and 20% acetonitrile, and buffer B consisted of 500 mM sodium perchlorate, 20 mM Tris, 1 mM EDTA, pH 7.4, and 20% acetonitrile. The desired product was isolated and desalted using a reversed-phase C18 column.

[0474] Annealing of single-stranded oligoribonucleotides to produce siRNA: Prepare the single-stranded oligoribonucleotides to be annealed at a concentration of 200 μM using sterile RNase-free water. Set up the annealing reaction as follows: Place 10 nmol of the mixture in a 95°C waterbath for 10 minutes (amounts ≥ 100 nmol require 20 minutes at high temperature). Immediately place the mixture in a 60°C waterbath and allow to cool naturally. Do not store the annealed solution at high temperatures. Complementary strands are formed by combining equimolar amounts of the single-stranded oligoribonucleotide solutions. The unmodified siRNA sequences are shown in Table 1.

[0475] The siRNA duplexes were sequence-modified and optimized, where lowercase letters "g", "c", "a" and "u" represent 2'-methoxy-modified nucleotides; uppercase letters "Gf", "Cf", "Af" and "Uf" represent 2'-fluoro-modified nucleotides; * indicates that the two monomers adjacent to the left and right (e.g., two nucleotides) are connected by a phosphorothioate group.

[0476] invAB refers to an abasic nucleotide with an inverted linkage at the 5' or 3' end of the nucleotide.

[0477] In the present invention, (M06) It is an M06 monomer In certain embodiments, (M06) is obtained by Bonded to a nucleotide.

[0478] In certain embodiments, [D02] is through Bonded to a nucleotide.

[0479] The modified siRNA sequences are shown in Table 2.

[0480] Example 2 In vitro effect detection of chemically modified duplex siRNA on SY5Y cells

[0481] The duplex siRNA selected according to Example 1 was further verified for its in vitro inhibitory effect on SY5Y cells.

[0482] 1) SY5Y cell transfection

[0483] SY5Y cells were cultured in DMEM supplemented with 10% fetal bovine serum, 1% glutamine, 1% NEAA, and 1% penicillin-streptomycin in a 5% CO2, 37°C incubator. Transfection was performed when the cells were in logarithmic growth phase and in good condition. SY5Y cells were plated in 96-well plates and transfected with siRNA using RNAiMAX according to the manufacturer's instructions. Final siRNA concentrations for this assay were 30 nM and 0.1 nM. A control group containing RNAiMAX and no compound was also established.

[0484] 2) RNA extraction and reverse transcription

[0485] 24 hours after transfection, the culture medium was removed and the cells were collected for RNA extraction. Total RNA was extracted using QIAGEN-74182 RT Kit and cDNA was synthesized using the FastKing RT Kit (with gDNase) (Tiangen-KR116-02) according to the manufacturer's instructions.

[0486] 3) qPCR detection of target gene mRNA expression level

[0487] Target cDNA will be detected by SYBR Green qPCR, with GAPDH cDNA used in parallel as an internal control. 8 μL of the prepared PCR reaction solution and 2 μL of sample cDNA will be added to a 384-well plate. The qPCR reaction program is as follows: 50°C for 2 minutes, 95°C for 10 minutes, followed by cycling at 95°C for 15 seconds, followed by 60°C for 1 minute, for a total of 40 cycles.

[0488] 4) Result analysis

[0489] The expression level of the target gene mRNA in each sample was calculated using the ΔΔCt relative quantification method. The relative amount of the target gene was expressed as 2-ΔΔCT.

[0490] The calculation formula is as follows:

[0491] ΔCT = average Ct value of target gene - average Ct value of reference gene

[0492] ΔΔCT = ΔCT (drug-treated group) - ΔCT (RNAiMAX control group)

[0493] Relative expression of target gene IGF-1R = 2-ΔΔCT

[0494] IGF-1R inhibition rate % = (1-value of sample / Ave.value of RNAiMAX Control) * 100

[0495] The results are shown in Table 3.

[0496] Table 3 Inhibitory effects of duplex siRNA on SY5Y (percent inhibition rate)

[0497] The results showed that the double-stranded siRNA could significantly inhibit the level of IGF-1R mRNA in SY5Y cells.

[0498] Example 3 In vitro detection of the effect of chemically modified duplex siRNA on Hela cells

[0499] The duplex siRNA selected according to Example 1 was further verified for its in vitro inhibitory effect on Hela cells.

[0500] 1) Hela cell transfection

[0501] HeLa cells were cultured in DMEM supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin in a 5% CO2, 37°C incubator. Transfection was performed when the cells were in logarithmic growth phase and in good condition. HeLa cells were seeded into 96-well plates and plated. Double-stranded siRNA was transfected into HeLa cells using lipofectamine RNAiMAX and Opti-MEM according to the manufacturer's instructions. Final siRNA concentrations used in this assay were 30 nM, 5 nM, and 0.5 nM.

[0502] 2) RNA extraction, reverse transcription, and qPCR

[0503] Total RNA was extracted using the EZ 96 Total RNA Kit (Omega, R1034-02) according to the kit instructions, and cDNA was synthesized using the FastKing RT Kit (With gDNase) (Tiangen-KR116-02) according to the kit instructions.

[0504] 3) qPCR detection of target gene mRNA expression level

[0505] The experimental steps are the same as in Example 2.

[0506] 4) Result analysis

[0507] The experimental result analysis process is the same as that of Example 2.

[0508] The results are shown in Table 4.

[0509] Table 4 Inhibition results of duplex siRNA on Hela (percent inhibition rate)

[0510] The results showed that the double-stranded siRNA could significantly inhibit the level of IGF-1R mRNA in Hela cells.

[0511] Example 4 In vitro detection of the effect of chemically modified duplex siRNA on ARPE cells

[0512] The duplex siRNA selected according to Example 1 was further verified for its in vitro inhibitory effect on ARPE cells.

[0513] 1) ARPE cell transfection

[0514] ARPE-19 cells were cultured in DMEM supplemented with 10% fetal bovine serum in a 5% CO2, 37°C incubator. Transfection was performed when the cells were in logarithmic growth phase and in good condition. ARPE-19 cells were seeded into 96-well plates and plated. HeLa cells were then transfected with double-stranded siRNA using lipofectamine RNAiMAX and Opti-MEM according to the manufacturer's instructions. Final siRNA concentrations used in this assay were 30 nM, 0.5 nM, and 0.05 nM.

[0515] 2) RNA extraction, reverse transcription, and qPCR

[0516] The experimental steps are the same as in Example 2.

[0517] 3) qPCR detection of target gene mRNA expression level

[0518] The experimental steps are the same as in Example 2.

[0519] 4) Result analysis

[0520] The experimental result analysis process is the same as that of Example 2.

[0521] The results are shown in Table 5.

[0522] Table 5 Inhibitory effects of duplex siRNA on ARPE (percent inhibition rate)

[0523] The results showed that the double-stranded siRNA could significantly inhibit the level of IGF-1R mRNA in ARPE cells.

[0524] Example 5 IC of chemically modified duplex siRNA in human primary mature adipocytes 50 Detection

[0525] The duplex siRNA selected according to Example 1 was further verified for its IC50 effect on mature human adipocytes.

[0526] 5) Transfection of primary human mature adipocytes

[0527] Primary human mature adipocytes were cultured in a 5% CO2, 37°C incubator in a specialized adipocyte culture medium. Transfection was performed when the cells were in logarithmic growth phase and in good condition. Mature adipocytes were seeded into 24-well plates and plated. Double-stranded siRNA was then transfected into the mature adipocytes using lipofectamine RNAiMAX and Opti-MEM according to the manufacturer's instructions.

[0528] 6) RNA extraction, reverse transcription, and qPCR

[0529] The experimental steps are the same as in Example 2.

[0530] 7) qPCR detection of target gene mRNA expression level

[0531] The experimental steps are the same as in Example 2.

[0532] 8) Result analysis

[0533] The results are shown in Table 6.

[0534] Table 6 Inhibitory effects of duplex siRNA on primary human mature adipocytes

[0535] The results showed that the double-stranded siRNA could significantly inhibit the level of IGF-1R mRNA in human primary mature adipocytes.

[0536] Example 6 PD study of intraorbital administration in cynomolgus monkeys

[0537] Group design

[0538] Six female cynomolgus macaques were divided into two groups. Prior to dosing, a biopsy was performed on the left eye, and a small amount of ocular muscle and periorbital fat was collected. The collected samples were placed separately in homogenized tubes containing 0.5 mL of RNAlater (two tubes for ocular muscle and one tube for fat) and placed in a refrigerator at 2°C–8°C for approximately 22–26 hours, after which the RNAlater was completely removed. The homogenized tubes containing the tissues were temporarily frozen on dry ice and then placed in a refrigerator at <-60°C. On the first day of the experiment, PBS was administered to the left eye, and different doses (2 mg / kg and 10 mg / kg) of the test article Z4 were administered to the right eye.

[0539] Test procedures

[0540] 1 Clinical observation

[0541] All animals underwent detailed clinical observation at least once before the experiment and once each on days 2, 7, 14, 21, and 28 after administration.

[0542] 2 weight

[0543] All animals were weighed at least once before the experiment and once on day 28 or on the day of planned necropsy.

[0544] 3. Ophthalmological examination

[0545] All animals underwent ophthalmological examination at least once before the experiment and once on days 2, 7, 14, 21, and 28 after administration.

[0546] 4. Intraocular pressure

[0547] The intraocular pressure (IOP) of both eyes of all animals was measured using a TonoVet tonometer before the start of drug administration and once on days 2 and 7 after drug administration.

[0548] 5 Fundus photography (FP)

[0549] Fundus photography (FP) was performed on both eyes of all animals at least once before the experiment and at week 4, and pictures were collected.

[0550] 6Flash visual evoked potential (fVEP)

[0551] All animals will undergo an fVEP examination in both eyes at week 4. Animals will be anesthetized with an intramuscular injection of ketamine (10-30 mg / kg) and xylazine (0.5 to 1.0 mg / kg). Mydriasis will be administered with an appropriate mydriatic agent. Flash VEP will be performed according to the following steps:

[0552] Select the FVEP protocol and enter the animal information;

[0553] Place the animal prone on a lifting platform and connect the relevant electrodes;

[0554] Check the resistance of each electrode. If it meets the requirements, click Start to collect electrophysiological signals.

[0555] After judging the inspection results, save the inspection report.

[0556] 7. Cytokine Analysis

[0557] Blood samples will be collected from all available animals for serum cytokine analysis. Blood samples will be collected before dosing and 24 hours after dosing. Serum levels of IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, TNF-α, and IFN-γ will be analyzed using validated flow cytometry.

[0558] Endpoint anatomy

[0559] All experimental animals were euthanized on day 29 and tissues were collected. During the tissue collection process, the eyeball, eye muscles (including the superior rectus, superior oblique, medial rectus, lateral rectus, inferior oblique, and inferior rectus muscles), and periorbital fat were grossly observed.

[0560] For all experimental animals, ocular muscles (including superior rectus, superior oblique, medial rectus, lateral rectus, inferior oblique and inferior rectus) and periorbital fat were collected from both eyes.

[0561] IGF1R mRNA QPCR analysis

[0562] IGF1R mRNA expression was measured in cynomolgus macaque tissues using a dye-based RT-qPCR method. The CT values ​​of the target gene (IGF1R) and the reference gene (CypA) were measured, and changes in target gene expression were calculated. Tissues included periocular muscle and periocular fat.

[0563] 1Key primer reagents

[0564] 2 key equipment

[0565] qPCR thermal cycler, PCR thermal cycler, biological safety cabinet, homogenizer, cryo-grinder, electronic balance

[0566] CyBio-SELMA liquid handling workstation, UV spectrophotometer, TECAN liquid handling workstation

[0567] 3. RNA extraction

[0568] Add 1000 μL of TRNzol Universal RNA Reagent to each tissue sample, followed by three 3 mm grinding beads. Set the homogenizer to 65 Hz, 45 s / cycle for four cycles, with a 30 s dwell between cycles (or 5.65 m / s, 1 min / cycle for three cycles, with a 30 s dwell between cycles) to obtain tissue lysates. Centrifuge each tissue lysate at 12,000 × g for 5 min at room temperature, and transfer all supernatants to new Eppendorf tubes. Incubate at room temperature for 5 minutes to completely dissociate the nucleoprotein complex. Add 200 μL of chloroform to each tissue lysate, vortex to mix, centrifuge briefly, and incubate at room temperature for 3 minutes. Centrifuge again at 4°C, 12,000 × g for 5 min. After centrifugation, the tissue lysate separates into three aqueous phases: upper, middle, and lower. Transfer 450 μL of the upper aqueous phase to a new Eppendorf tube. Add 45 μL of 3M Sodium Acetate Sol (pH 5.2) to each aliquot, vortex to mix, and incubate at room temperature for 1 minute. After incubation, add 500 μL of pre-chilled isopropanol to each aliquot, vortex to mix, and incubate at -30°C to -10°C for 30 minutes. After incubation, centrifuge at 4°C, 12,000 × g, and 15 minutes. If RNA precipitates are visible at the bottom of the tube, discard the supernatant. After discarding the supernatant, add 1000 μL of pre-chilled 75% ethanol to each aliquot, vortex to mix, and centrifuge at 4°C, 12,000 × g, and 10 minutes. After centrifugation, discard the supernatant, air-dry at room temperature for 10 minutes, then add 40 μL of DEPC-treated water (pre-heated at 55°C) to each aliquot and vortex to mix. RNA purity and concentration were determined using a UV spectrophotometer using A260 / A280 and recorded. RNA can be used directly for RT-qPCR or stored at <-60°C. The expiration date of RNA is 1 year.

[0569] 4QPCR analysis

[0570] RNA was run through a genomic DNA removal program to generate the "RNA template (DNA removed)." This was then run through a PCR thermal cycler program to generate cDNA products, which served as the "qPCR template." All cDNA samples were analyzed in triplicate using a 384-well plate for qPCR. Relative expression data were collected using a QuantStudio7 Flex PCR system, including mean Ct, ΔCt, and ΔΔCt. KD% values ​​for all IGF1R mRNAs were calculated after normalization to biopsy samples.

[0571] Experimental results:

[0572] 1. Clinical observation, body weight, intraocular pressure, fundus photography, flash visual evoked potential (fVEP), and cytokine analysis showed no abnormalities;

[0573] 2. KD% experimental results: as shown in Figure 1.

[0574] Experimental conclusion: The double-chain compound Z4 has good safety when administered intraorbitally and significantly inhibits the expression of IGF1R mRNA in target tissues.

[0575] Example 7: Acute exophthalmos model in mice

[0576] Objective: To evaluate the efficacy of mouse IGF1R siRNA Z8 and Z9 delivered intraorbitally in a BALB / c mouse thyroid eye disease model.

[0577] 1. Group dosing regimen

[0578] On day 5, the animals were anesthetized with Zotal (25-50 mg / kg, ip) and xylazine hydrochloride (5 mg / kg, ip), and a single intraorbital injection of mouse IGF1R siRNA was performed on both eyes at 100 μL / eye.

[0579] 2 Modeling

[0580] On Day 0 and Day 6, the animals were anesthetized with Zotai (25-50 mg / kg, ip) and xylazine hydrochloride (5 mg / kg, ip), and mouse IGF-1 protein was injected periocularly to establish the model at 5 μg / eye. After injection, both eyes of the animals were cared for with levofloxacin eye drops and ofloxacin eye ointment once in the morning and once in the afternoon for three consecutive days.

[0581] 3. Detection indicators

[0582] 3.1 Clinical Observation

[0583] Once a day, observe the animal's condition, mental state, behavioral activities, eating habits, etc. next to the cage.

[0584] 3.2 GO (Graves' Ophthalmopathy) score

[0585] GO scores were performed on Day 0 (before modeling), Day 3, Day 6 (before modeling), and Day 9, and the symptoms of proptosis were continuously observed.

[0586] 3.3 Eye examination

[0587] Ocular surface and fundus examinations were performed using a slit lamp on Day 5 (before and after administration), Day 0, Day 3, Day 7, and Day 14. Any abnormalities were photographed and recorded.

[0588] 3.4 Collection of materials

[0589] On day 9, four animals from each group were euthanized, and the periorbital tissue, retina, sclera complex, liver, skeletal muscle, and ovarian fat were isolated and immediately stored in RNAprotect tissue Reagent.

[0590] 3.5 qPCR detection

[0591] RNA was extracted from periorbital tissues and subjected to reverse transcription and QPCR analysis.

[0592] 3.6 Pathology

[0593] On Day 9, eyeballs (with periocular tissues) of 4 mice in each group were fixed and embedded in paraffin, and the proliferation and cell infiltration of periocular tissues were quantified (H&E staining, UCPI and CD3 staining).

[0594] Intraorbital administration of mouse IGF1R siRNA can significantly inhibit the expression of IGF1R mRNA and alleviate the symptoms of exophthalmos, indicating that intraorbital administration of the compounds of the present invention can effectively treat thyroid eye disease.

[0595] Experimental Example 7: Rat surgically induced osteoarthritis model

[0596] Ten weeks after birth, 24 rats underwent surgery to transcribe the anterior cruciate ligament, medial collateral ligament, and medial meniscocondylar ligament (anterior cruciate ligament rupture and partial medial meniscectomy model; ACLT+pMMx). One week after surgery, all rats were randomly assigned to receive intra-articular injection of mouse IGF1R siRNA Z8, Z9, or control solution (12 rats per group). 13 weeks after surgery, the knee joints were isolated, fixed in 10% formalin, decalcified, and embedded in paraffin blocks. Anterior sections (5 μm thickness, 100 μm interval between each section to ensure reproducibility) were taken from different levels and stained with safranin fast green. At least 12 sections were taken from each rat and imaged using a light microscope.

[0597] Osteoarthritis Research Society International (OARSI) score, cartilage protection and regeneration

[0598] Histological evaluation was performed by two blinded observers. Images were scored according to the OARSI cartilage histology scoring system, which assesses the extent of cartilage damage (depth of cartilage damage) and stage (extent of joint involvement). Briefly, the femur and tibia were evaluated separately, and each site was scored based on the grade of cartilage damage (0–6, with 0 indicating an intact surface and 6 indicating severe deformation) and the stage of cartilage damage (0–4, with 0 indicating a normal joint and 4 indicating greater than 50% joint damage). The total score was the product of the grade and stage (0 representing a normal joint and 24 representing severe osteoarthritis). Twelve sections were scored per rat, and no rats were excluded. After scoring, the study was unblinded, and the four lowest-scoring sections (representing the least damaged sections) from each rat were excluded from further analysis. The mean OARSI score for each rat was the average of the scores of the two blinded observers. In addition, four independent blinded observers repeated the histological evaluation based on the revised objective quantitative histological OARSI scoring system. Safranin O staining intensity and cartilage thickness were measured using ImageJ.

[0599] Intra-articular administration of mouse IGF1R siRNA can significantly inhibit the expression of IGF1R mRNA and reduce the OARSI score, indicating that intra-articular administration of the compounds of the present invention can effectively treat osteoarthritis in humans.

[0600] Example 8: Spinal Nerve Ligation (SNL) Model

[0601] Animals were first anesthetized with 5% isoflurane and maintained under 2% isoflurane. The L5 transverse process was removed to expose the L3 and L4 spinal nerves. The L4 spinal nerve was then isolated and tightly ligated with 6-0 silk suture. For the sham-operated group, the L4 spinal nerve was exposed but not ligated. Under isoflurane anesthesia, mice were injected with mouse IGF1R siRNA Z8, Z9, or a control solution via lumbar puncture, and subsequent behavioral testing was performed.

[0602] Behavioral testing

[0603] 50% paw withdrawal mechanical threshold (PWT):

[0604] Acclimation period: The rats were acclimated to the test area for at least three days before measurements were taken. Vertical stimulation of the lateral plantar aspect was performed using von Frey fibers of varying strengths (0.6 g, 1.0 g, 1.4 g, 2.0 g, 4.0 g, 6.0 g, 8.0 g, 10.0 g, and 15.0 g). Measurements were performed one day before surgery and on the 4th, 7th, 10th, and 14th days after surgery. First, the 4.0 g fiber was selected, and then the 50% PWT threshold of the rat was calculated using the "up-down method." The proportion of paw retraction in response to increasing mechanical stimulation was measured. Three groups of rats were evaluated sequentially, with five measurements performed on each side (5 minutes apart each time). Finally, the five paw retraction response rates, i.e., the proportion of paw retraction in response to a 6.0 g mechanical stimulus, were calculated.

[0605] Cold hyperalgesia threshold:

[0606] To assess the sensitivity of the paw to cold, the rat's paw response to acetone was measured. While the rat was resting, pre-cooled acetone (100 μL) was applied to the plantar surface of the paw. The duration of a positive response (lifting the paw, shaking the paw, licking, etc.) was recorded for 30 seconds. Each paw was tested three times (5 minutes apart), and the average of the three measurements was recorded as the reaction time for that side.

[0607] Intrathecal administration of mouse IGF1R siRNA can significantly inhibit the expression of IGF1R mRNA, while improving PWT and cold hyperalgesia threshold scores, and inhibiting pain response, suggesting that intrathecal administration of the compounds involved in the present invention can effectively treat neuropathic pain in humans.

Claims

1. An RNA inhibitor that inhibits the expression of insulin-like growth factor 1 receptor (IGF-1R) gene, comprising an antisense strand that forms a complementary region with at least 15 consecutive nucleotides in the mRNA (SEQ ID NO: 828) encoding IGF-1R, and the complementary region has 0, 1, 2, 3, 4 or 5 mismatches. Preferably, the length of the complementary region is 15 - 30 nucleotide pairs, and more preferably 17 - 23 nucleotide pairs.

2. The RNA inhibitor according to claim 1, wherein the antisense strand forms a complementary region with any one of the following consecutive 15, 16, 17, 18, 19, 20, 21, 22 or 23 nucleotides starting from the 5'-end in the mRNA (SEQ ID NO: 828) encoding IGF-1R: The 1118th nucleotide, the 1120th nucleotide, the 1154th nucleotide, the 1367th nucleotide, the 1407th nucleotide, the 1415th nucleotide, the 1532nd nucleotide, the 1625th nucleotide, the 1627th nucleotide, the 1628th nucleotide, the 1631st nucleotide, the 3356th nucleotide, the 3357th nucleotide, the 3359th nucleotide, the 3792nd nucleotide, the 4198th nucleotide, the 4200th nucleotide, the 4208th nucleotide, the 4685th nucleotide, the 5246th nucleotide, the 5392nd nucleotide, the 5393rd nucleotide, the 6329th nucleotide, the 6332nd nucleotide, the 6333rd nucleotide, the 6334th nucleotide, the 10215th nucleotide, the 10738th nucleotide, the 10740th nucleotide, the 10770th nucleotide, the 10772nd nucleotide, the 10773rd nucleotide, the 10946th nucleotide, the 10956th nucleotide, the 10957th nucleotide, the 1418th nucleotide, the 1541st nucleotide, the 2050th nucleotide, the 2055th nucleotide, the 2056th nucleotide, the 2233rd nucleotide, the 2234th nucleotide, the 2456th nucleotide, the 2591st nucleotide, the 2603rd nucleotide, the 2606th nucleotide, the 2609th nucleotide, the 2776th nucleotide, the 2890th nucleotide, the 2906th nucleotide, the 2909th nucleotide, the 2957th nucleotide, the 2960th nucleotide, the 3008th nucleotide, the 3195th nucleotide, the 3398th nucleotide, the 3608th nucleotide, the 3609th nucleotide, the 3650th nucleotide, the 3660th nucleotide, the 3815th nucleotide, the 3882nd nucleotide, the 3996th nucleotide, the 4067th nucleotide, the 4121st nucleotide, the 4122nd nucleotide, the 4292nd nucleotide, the 4293rd nucleotide, the 4295th nucleotide, the 4516th nucleotide, the 4519th nucleotide, the 5163rd nucleotide, the 5309th nucleotide, the 5310th nucleotide, the 5377th nucleotide, the 5380th nucleotide, the 5381st nucleotide, the 5663rd nucleotide, the 5700th nucleotide, the 5711th nucleotide, the 5712th nucleotide, the 5721st nucleotide, the 5896th nucleotide, the 5898th nucleotide, the 6203rd nucleotide, the 6236th nucleotide, the 6237th nucleotide, the 6240th nucleotide, the 6245th nucleotide, the 6288th nucleotide, the 6299th nucleotide, the 6305th nucleotide, the 6309th nucleotide, the 6310th nucleotide, the 6343rd nucleotide, the 6363rd nucleotide, the 6365th nucleotide, the 6366th nucleotide, the 6373rd nucleotideThe 6379th nucleotide, the 6381st nucleotide, the 6431st nucleotide, the 6434th nucleotide, the 6496th nucleotide, the 6518th nucleotide, the 6755th nucleotide, the 6760th nucleotide, the 6883rd nucleotide, the 6885th nucleotide, the 6947th nucleotide, the 6948th nucleotide, the 6949th nucleotide, the 6952nd nucleotide, the 6953rd nucleotide, the 6954th nucleotide, the 7574th nucleotide, the 7621st nucleotide, the 7764th nucleotide, the 7830th nucleotide, the 7906th nucleotide, the 8540th nucleotide, the 8836th nucleotide, the 8892nd nucleotide, the 8975th nucleotide, the 8977th nucleotide, the 9163rd nucleotide, the 9175th nucleotide, the 9186th nucleotide, the 9265th nucleotide, the 9267th nucleotide, the 9340th nucleotide, the 9347th nucleotide, the 9352nd nucleotide, the 9356th nucleotide, the 9478th nucleotide, the 9480th nucleotide, the 9606th nucleotide, the 9611th nucleotide, the 9634th nucleotide, the 9680th nucleotide, the 9681st nucleotide, the 9724th nucleotide, the 9725th nucleotide, the 9814th nucleotide, the 10015th nucleotide, the 10016th nucleotide, the 10189th nucleotide, the 10244th nucleotide, the 10266th nucleotide, the 10490th nucleotide, the 10526th nucleotide, the 10528th nucleotide, the 10534th nucleotide, the 10535th nucleotide, the 10537th nucleotide, the 10540th nucleotide, the 10541st nucleotide, the 10679th nucleotide, the 10694th nucleotide, the 10717th nucleotide, the 10728th nucleotide, the 10729th nucleotide, the 10730th nucleotide, the 10754th nucleotide, the 10763rd nucleotide, the 10766th nucleotide, the 10942nd nucleotide, the 11034th nucleotide, the 11078th nucleotide, the 11126th nucleotide, the 11156th nucleotide, the 11159th nucleotide, the 11160th nucleotide, the 11292nd nucleotide, the 11340th nucleotide, the 11344th nucleotide, the 11383rd nucleotide, the 11395th nucleotide, the 11789th nucleotide, the 11864th nucleotide, the 11865th nucleotide, the 11939th nucleotide, the 12092nd nucleotide, the 12116th nucleotide, the 12130th nucleotide, the 12154th nucleotide, the 12156th nucleotide, the 12177th nucleotide, the 12195th nucleotide, the 1164th nucleotide, the 1163rd nucleotide, the 1162nd nucleotide, the 1161st nucleotide, the 1160th nucleotideThe 1165th nucleotide, the 1166th nucleotide, the 1167th nucleotide, the 1168th nucleotide.

3. The RNA inhibitor according to any one of claims 1 - 2, wherein the sequence between the 1160th nucleotide and the 1188th nucleotide, the sequence between the 6235th nucleotide and the 6465th nucleotide, or the sequence between the 6329th nucleotide and the 6455th nucleotide starting from the 5'-end in the mRNA (SEQ ID NO: 828) encoding IGF-1R forms a complementary region with the antisense strand.

4. The RNA inhibitor according to any one of claims 1 - 3, wherein the RNA inhibitor is ribonucleic acid. Further, the RNA inhibitor is single-stranded ribonucleic acid or double-stranded ribonucleic acid.

5. The RNA inhibitor according to any one of claims 1 - 4, wherein the RNA inhibitor is antisense oligonucleotide (ASO), shRNA, miRNA or siRNA.

6. The RNA inhibitor according to any one of claims 1 - 5, which comprises a sense strand capable of forming a complementary double strand with the antisense strand, wherein the antisense strand contains a sequence that forms a duplex complementary region with at least 15 consecutive nucleotides in the sense strand sequence, and the duplex complementary region has 0, 1, 2, 3, 4 or 5 mismatches. Preferably, the length of the duplex complementary region is 15 - 30 nucleotide pairs, and more preferably 17 - 23 nucleotide pairs.

7. The RNA inhibitor according to claim 6, wherein the sense strand and the antisense strand are present on two different nucleic acid strands. Preferably, the RNA inhibitor is siRNA or shRNA, and more preferably the RNA inhibitor is siRNA.

8. The RNA inhibitor according to any one of claims 6 - 7, wherein the sense strand and the antisense strand are present on the same nucleic acid strand. Preferably, the RNA inhibitor is shRNA.

9. The RNA inhibitor according to any one of claims 6 - 8, wherein the total length of the sense strand is 15 - 50 nucleotides. Preferably, the total length of the sense strand is 16 - 30 nucleotides, and more preferably the total length of the sense strand is 17, 18, 19, 20 or 21 nucleotides.

10. The RNA inhibitor according to any one of claims 6 to 9, wherein the total length of the antisense strand is 19-50 nucleotides, preferably the total length of the antisense strand is 19-30 nucleotides, and more preferably the total length of the antisense strand is 21, 22, 23, 24, 25, 26 or 27 nucleotides.

11. The RNA inhibitor according to any one of claims 6 to 10, wherein the sense strand and the antisense strand each independently optionally comprise a 3' or 5' overhang of 1, 2 or 3 nucleotides.

12. The RNA inhibitor according to claim 11, wherein both the sense strand and the antisense strand have a 3' overhang of 1-3 nucleotides in length, or the sense strand has a 3' or 5' overhang of 1-3 nucleotides in length, or the antisense strand has a 3' or 5' overhang of 1-3 nucleotides in length.

13. The RNA according to any one of claims 1-12, wherein the antisense strand comprises at least 15 consecutive nucleotides in the sequence of any one of SEQ ID NOs: 199-400.

14. The RNA inhibitor according to any one of claims 6 to 13, wherein the sense strand comprises at least 15 consecutive nucleotides in the sequence of any one of SEQ ID NOs: 1 to 198.

15. The RNA inhibitor according to any one of claims 1-14, which comprises a duplex selected from the following: ds-n1, ds-n2, ds-n3, ds-n4, ds-n5, ds-n6, ds-n7, ds-n8, ds-n9, ds-n10, ds-n11, ds-n12, ds-n13, ds-n14, ds-n15, ds-n16, ds-n17, ds-n18, ds-n19, ds-n20, ds-n21, ds-n22, ds-n23, ds-n24, ds-n25, ds-n26, ds-n27, ds-n28, ds-n29, ds-n30, ds-n31, ds-n32, ds-n33, ds-n34, ds-n35, ds-n36, ds-n37, ds-n38, ds-n39, ds-n40, ds-n41, ds-n42, ds-n43, ds-n44, ds-n45, ds-n46, ds-n47, ds-n48, ds-n49, ds-n50, ds-n51, ds-n52, ds-n53, ds-n54, ds-n55, ds-n56, ds-n57, ds-n58, ds-n59, ds-n60, ds-n61, ds-n62, ds-n63, ds-n64, ds-n65, ds-n66, ds-n67, ds-n68, ds-n69, ds-n70, ds-n71, ds-n72, ds-n73, ds-n74, ds-n75, ds-n76, ds-n77, ds-n78, ds-n79, ds-n80, ds-n81, ds-n82, ds-n83, ds-n84, ds-n85, ds-n86, ds-n87, ds-n88, ds-n89, ds-n90, ds-n91, ds-n92, ds-n93, ds-n94, ds-n95, ds-n96, ds-n97, ds-n98, ds-n99, ds-n100, ds-n101, ds-n102, ds-n103, ds-n104, ds-n105, ds-n106, ds-n107, ds-n108, ds-n109, ds-n110, ds-n111, ds-n112, ds-n113, ds-n114, ds-n115, ds-n116, ds-n117, ds-n118, ds-n119, ds-n120, ds-n121, ds-n122, ds-n123, ds-n124, ds-n125, ds-n126, ds-n127, ds-n128, ds-n129, ds-n130, ds-n131, ds-n132, ds-n133,ds-n134, ds-n135, ds-n136, ds-n137, ds-n138, ds-n139, ds-n140, ds-n141, ds-n142, ds-n143, ds-n144, ds-n145, ds-n146, ds-n147, ds-n148, ds-n149, ds-n150, ds-n151, ds-n152, ds-n153, ds-n154, ds-n155, ds-n156, ds-n157, ds-n158, ds-n159, ds-n160, ds-n161, ds-n162, ds-n163, ds-n164, ds-n165, ds-n166, ds-n167, ds-n168, ds-n169, ds-n170, ds-n171, ds-n172, ds-n173, ds-n174, ds-n175, ds-n176, ds-n177, ds-n178, ds-n179, ds-n180, ds-n181, ds-n182, ds-n183, ds-n184, ds-n185, ds-n186, ds-n187, ds-n188, ds-n189, ds-n190, ds-n191, ds-n192, ds-n193, ds-n194, ds-n195, ds-n196, ds-n197, ds-n198, ds-n201, ds-n202, ds-n203, ds-n204。、 16. The RNA inhibitor of claim 15, comprising a duplex selected from the group consisting of ds-n17, ds-n23, ds-n31, ds-n39, ds-n45, ds-n58, ds-n60, ds-n64, ds-n67, ds-n68, ds-n73, ds-n74, ds-n76, ds-n77, ds-n80, ds-n81, ds-n83, ds-n84, ds-n86, ds-n88, ds-n89, ds-n 98. ds-n99, ds-n103, ds-n104, ds-n112, ds-n113, ds-n115, ds-n153, ds-n154, ds-n156, ds-n157, ds-n163, ds- n164, ds-n165, ds-n188, ds-n190, ds-n191, ds-n192, ds-n193, ds-n194, ds-n195, ds-n196, ds-n197, ds-n198.

17. The RNA inhibitor according to claim 15, which comprises a duplex selected from the following: ds-n17, ds-n23, ds-n31, ds-n39, ds-n45, ds-n58, ds-n60, ds-n64, ds-n67, ds-n68, ds-n73, ds-n74, ds-n76, ds-n77, ds-n80, ds-n81, ds-n83, ds-n84, ds-n86, ds-n88, ds-n89, ds-n98, ds-n99, ds-n103, ds-n104, ds-n112, ds-n113, ds-n115, ds-n153, ds-n154, ds-n156, ds-n157, ds-n163, ds-n164, ds-n165, ds-n188.

18. The RNA inhibitor according to claim 16, which comprises a duplex selected from the following: ds-n23, ds-n58, ds-n67, ds-n86, ds-n89, ds-n98, ds-n99, ds-n103, ds-n190, ds-n198, ds-n113.

19. The RNA inhibitor according to any one of claims 1-18, characterized in that, The antisense strand further comprises region B1 at the 3'-end, and the sense strand further comprises region A1 at the 5'-end, wherein region A1 is 0 - 6 nucleotides and region B1 is 0 - 6 nucleotides.

20. The RNA inhibitor according to claim 19, wherein Region B1 is 0, 1, 2, 3, 4, 5 or 6 nucleotides; preferably, region B1 is 0 or 2 nucleotides.

21. The RNA inhibitor according to any one of claims 16-20, characterized in that, Region A1 is 0, 1, 2, 3, 4, 5 or 6 nucleotides; preferably, region A1 is 0 or 2 nucleotides.

22. The RNA inhibitor according to claim 21, wherein The antisense strand further comprises region X2 at the 5'-end, and the sense strand or the sense nucleic acid further comprises region X1 at the 3'-end, and region X1 and region X2 are complementary and paired.

23. The RNA inhibitor according to claim 22, wherein X1 is A, U, modified A or modified U, and X2 is U, A, modified U or modified A.

24. The RNA inhibitor according to any one of claims 19-23, characterized in that, The antisense strand sequentially comprises X2, Y, Z and N at the 5'-end in the 3' to 5' direction, wherein X2 and Y are each independently A, U, modified A or modified U, Z is G or modified G, and N comprises at least one nucleotide.

25. The RNA inhibitor according to any one of claims 19 - 24, wherein X2, Y, Z are sequentially AAG, AUG, UUG or UAG in the 3' to 5' direction, or the above sequences that are partially or fully modified.

26. The RNA inhibitor according to any one of claims 21 - 25, wherein X2, Y, Z are sequentially AAG, AUG, UUG or UAG in the 3' to 5' direction, or the above sequences that are partially or fully modified.

27. The RNA inhibitor according to any one of claims 21 - 26, wherein N is C or modified C.

28. The RNA inhibitor according to any one of claims 21-27, wherein X2, Y, Z, and N are AAGC and UAGC in sequence in the 3' to 5' direction, or the above sequences that are partially or fully modified.

29. The RNA inhibitor according to any one of claims 16-28, wherein the antisense strand comprises at least 15 consecutive nucleotides in the sequence according to any one of SEQ ID NOs: 401-413.

30. The RNA inhibitor according to any one of claims 1-29, which is selected from ds-n190-1, ds-n191-1, ds-n192-1, ds-n193-1, ds-n194-1, ds-n195-1, ds-n196-1, ds-n197-1, ds-n198-1, ds-n201-1, ds-n202-1, ds-n203-1, ds-n204-1.

31. The RNA inhibitor according to any one of claims 1-30, wherein at least one nucleotide in the RNA inhibitor is a modified nucleotide.

32. The RNA inhibitor according to any one of claims 1-31, wherein at least 70%, 80%, 90%, 95% of the nucleotides in the RNA inhibitor are modified nucleotides; preferably, all nucleotides are modified nucleotides.

33. The RNA inhibitor according to any one of claims 31-32, wherein the modification comprises one or more combinations of the following: 2'-OMe (2'-O-methyl) modification, 2'-F (2'-deoxy-2'-fluoro) modification, 2'-O-MOE (2'-O-methoxyethyl) modification, 2'-deoxy (2'-d) modification, 5'-morpholine (5'-Mo) modification, unlocked nucleic acid (UNA) modification, glycol nucleic acid (GNA) modification, locked nucleic acid (LNA) modification, tricyclic DNA (tcDNA) modification, (S)-constrained ethyl bicyclic nucleic acid ((S)-cEt-BNA) modification, phosphorothioate (PS) modification, dithiophosphorothioate (PS2) modification, methylphosphonate (MP) modification, methoxypropylmethylphosphonate (MOP) modification, peptide nucleic acid (PNA) modification, 5'-(E)-vinyl phosphonate (VP) modification (VP), N6-methyladenosine (m6A) modification, 5-methylcytidine (m5C) modification, 3-methyluridine (m3U) modification, 5-methyluridine (m5U) modification, pseudouridine modification, 2-thiouridine (s2U) modification, propynyluridine (5-pU) modification, modification of the abasic nucleotide with the 5'- or 3'-end bond of the nucleotide reversed (invAB) modification, replacement of the nucleotide with the abasic nucleotide with the reverse configuration (invAb) modification, replacement of the nucleotide with 2,4-difluorotolyl ribonucleotide (rF) modification or replacement of the nucleotide with (S)-glycerol nucleic acid modification, and the preferred modifications are 2'-OMe modification, 2'-F modification, 2'-deoxy modification, VP modification, 5'-MP modification, PS modification, PS2 modification, MP modification, MOP modification, M06 modification, invAb modification or invAB modification.

34. The RNA inhibitor according to any one of claims 31-33, wherein the 3'- or 5'-end of the antisense strand comprises a phosphate or a phosphate mimic, or the 3'- or 5'-end of the sense strand comprises a phosphate or a phosphate mimic.

35. The RNA inhibitor according to claim 34, wherein the phosphate mimic comprises 5'-(E)-vinyl phosphonate, 5'-methyl phosphonate, (S)-5'-C-methyl analogue and 5'-thiophosphate (5'-PS).

36. The RNA inhibitor according to any one of claims 31-35, wherein the 5'-end of the antisense strand comprises a (M06) modification:

37. The RNA inhibitor according to any one of claims 31-36, wherein the invAB modification is present at the 3'- or 5'-end of the antisense strand, or the invAB modification is present at the 3'- or 5'-end of the sense strand.

38. The RNA inhibitor according to any one of claims 31-37, wherein the abasic nucleotide with the reverse configuration or M06 is linked to the 3'- or 5'-end of the antisense strand or the sense strand through a phosphorothioate.

39. The RNA inhibitor according to any one of claims 31-38 has the following modification combination a) The sense strand contains a first sequence that is 16 - 21 nt in length, and the first sequence includes the following modifications: starting from the 5'-end and counting, the nucleotides at positions 9, 10, and 11 have 2'-F modifications; b) The antisense strand contains a second sequence that is 16 - 21 nt in length, and the second sequence includes the following modifications: there is a phosphorothioate linkage between the nucleotides at positions 1 and 2, a phosphorothioate linkage between the nucleotides at positions 2 and 3, and a phosphorothioate linkage between the nucleotides at positions 3 and 4, starting from the 5'-end and counting; and there is a phosphorothioate linkage between the nucleotides at positions 1 and 2, starting from the 3'-end and counting; and starting from the 5'-end and counting, the nucleotides at positions 2, 3, 4, 12, 14, and 16 have 2'-F modifications.

40. The RNA inhibitor according to claim 39, wherein the length of the first sequence is preferably 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, more preferably 21 nt.

41. The RNA inhibitor according to claim 39, wherein the length of the second sequence is preferably 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, more preferably 21 nt.

42. The RNA inhibitor according to any one of claims 39 - 41 has the following combination of modifications a) The sense strand has a length of 21 nt and includes the following modifications: starting from the 5'-end and counting, the nucleotides at positions 9, 10, and 11 have 2'-F modifications; b) The antisense strand has a length of 21 nt and includes the following modifications: there is a phosphorothioate linkage between the nucleotides at positions 1 and 2, a phosphorothioate linkage between the nucleotides at positions 2 and 3, and a phosphorothioate linkage between the nucleotides at positions 3 and 4, starting from the 5'-end and counting; and there is a phosphorothioate linkage between the nucleotides at positions 1 and 2, starting from the 3'-end and counting; and starting from the 5'-end and counting, the nucleotides at positions 2, 3, 4, 12, 14, and 16 have 2'-F modifications.

43. The RNA inhibitor according to any one of claims 28 - 42, wherein the antisense strand contains at least 15 consecutive nucleotides from any one of the sequences as set forth in SEQ ID NO: 624 - 812.

44. The RNA inhibitor according to any one of claims 28 - 42, wherein the sense strand contains at least 15 consecutive nucleotides from any one of the sequences as set forth in SEQ ID NO: 414 - 602.

45. The RNA inhibitor according to any one of claims 28 - 44 is selected from: ds-m1, ds-m2, ds-m3, ds-m4, ds-m5, ds-m6, ds-m7, ds-m8, ds-m9, ds-m10, ds-m11, ds-m12, ds-m13, ds-m14, ds-m15, ds-m16, ds-m17, ds-m18, ds-m19, ds-m20, ds-m21, ds-m22, ds-m23, ds-m24, ds-m25, ds-m26, ds-m27, ds-m28, ds-m29, ds-m30, ds-m31, ds-m32, ds-m33, ds-m34, ds-m35, ds-m36, ds-m37, ds-m38, ds-m39, ds-m40, ds-m41, ds-m42, ds-m43, ds-m44, ds-m45, ds-m46, ds-m47, ds-m48, ds-m49, ds-m50, ds-m51, ds-m52, ds-m53, ds-m54, ds-m55, ds-m56, ds-m57, ds-m58, ds-m59, ds-m60, ds-m61, ds-m62, ds-m63, ds-m64, ds-m65, ds-m66, ds-m67, ds-m68, ds-m69, ds-m70, ds-m71, ds-m72, ds-m73, ds-m74, ds-m75, ds-m76, ds-m77, ds-m78, ds-m79, ds-m80, ds-m81, ds-m82, ds-m83, ds-m84, ds-m85, ds-m86, ds-m87, ds-m88, ds-m89, ds-m90, ds-m91, ds-m92, ds-m93, ds-m94, ds-m95, ds-m96, ds-m97, ds-m98, ds-m99, ds-m100, ds-m101, ds-m102, ds-m103, ds-m104, ds-m105, ds-m106, ds-m107, ds-m108, ds-m109, ds-m110, ds-m111, ds-m112, ds-m113, ds-m114, ds-m115, ds-m116, ds-m117, ds-m118, ds-m119, ds-m120, ds-m121, ds-m122, ds-m123, ds-m124, ds-m125, ds-m126, ds-m127, ds-m128, ds-m129, ds-m130, ds-m131, ds-m132, ds-m133, ds-m134, ds-m135, ds-m136, ds-m137, ds-m138ds-m139, ds-m140, ds-m141, ds-m142, ds-m143, ds-m144, ds-m145, ds-m146, ds-m147, ds-m148, ds-m149, ds-m150, ds-m151, ds-m152, ds-m153, ds-m154, ds-m155, ds-m156, ds-m157, ds-m158, ds-m159, ds-m160, ds-m161, ds-m162, ds-m163, ds-m164, ds-m165, ds-m166, ds-m167, ds-m168, ds-m169, ds-m170, ds-m171, ds-m172, ds-m173, ds-m174, ds-m175, ds-m176, ds-m177, ds-m178, ds-m179, ds-m180, ds-m181, ds-m182, ds-m183, ds-m184, ds-m185, ds-m186, ds-m187, ds-m188, ds-m189。、 46. The RNA inhibitor according to any one of claims 28 - 39, wherein a) The sense strand contains a third sequence with a length of 18 - 21 nt, and the third sequence includes the following modifications: starting from the 5'-end and counting, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, a phosphorothioate linkage between the 2nd and 3rd nucleotides, the nucleotides at positions 9, 10, 11, and 18 of the sense strand have 2'-F modification, and the rest have 2'-OMe modification; starting from the 3'-end and counting, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and a phosphorothioate linkage between the 2nd and 3rd nucleotides. b) The antisense strand contains a fourth sequence with a length of 19 - 26 nt, and the fourth sequence includes the following modifications: starting from the 5'-end and counting, the nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense strand have 2'-F modification, the rest have 2'-OMe modification, and there is a phosphorothioate linkage between the 4th and 5th nucleotides; starting from the 3'-end and counting, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and a phosphorothioate linkage between the 2nd and 3rd nucleotides.

47. The RNA inhibitor according to claim 46, wherein the length of the third sequence is preferably 18 nt, 19 nt, 20 nt, 21 nt, 22 nt, 21 nt, more preferably 21 nt.

48. The RNA inhibitor according to claim 47, wherein the length of the fourth sequence is preferably 19 nt, 20 nt, 21 nt, 22 nt, 23 nt, 24 nt, 25 nt, 26 nt, more preferably 26 nt.

49. The RNA inhibitor according to any one of claims 46 - 48, wherein a) The sense strand has a length of 21 nt. Starting from the 5'-end and counting, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, a phosphorothioate linkage between the 2nd and 3rd nucleotides, the nucleotides at positions 9, 10, 11, and 18 of the sense strand have 2'-F modification, and the rest have 2'-OMe modification; starting from the 3'-end and counting, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and a phosphorothioate linkage between the 2nd and 3rd nucleotides. b) The antisense strand has a length of 26 nt. Starting from the 5'-end and counting, the nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense strand have 2'-F modification, the rest have 2'-OMe modification, and there is a phosphorothioate linkage between the 4th and 5th nucleotides; starting from the 3'-end and counting, there is a phosphorothioate linkage between the 1st and 2nd nucleotides, and a phosphorothioate linkage between the 2nd and 3rd nucleotides.

50. The RNA inhibitor according to any one of claims 31 - 49, wherein the antisense strand contains at least 15 consecutive nucleotides from any one of the sequences as set forth in SEQ ID NO: 813 - 827.

51. The RNA inhibitor according to any one of claims 31 - 50, wherein the sense strand contains at least 15 consecutive nucleotides from any one of the sequences as set forth in SEQ ID NO: 603 - 617.

52. The RNA inhibitor according to any one of claims 31 - 51, which comprises a duplex selected from the following: ds-m190, ds-m191, ds-m192, ds-m193, ds-m194, ds-m195, ds-m196, ds-m197, ds-m198, ds-m199, ds-m200, ds-m201, ds-m202, ds-m203, and ds-m204.

53. The RNA inhibitor according to any one of claims 1 - 52, further comprising a delivery system, wherein the delivery system is conjugated to the sense strand and / or the antisense strand, and the delivery system is capable of enabling the RNA inhibitor to reach the target RNA in the target tissue to produce a gene silencing effect.

54. The RNA inhibitor according to claim 53, wherein, The target tissue is ocular tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, adipose tissue.

55. The RNA inhibitor according to claim 54, wherein, The ocular tissue is optic nerve, trabecular meshwork, juxtacanalicular tissue, ganglion, episcleral vein, Schlemm's canal, or peripheral ocular tissue. Preferably, the joint tissue comprises cartilage tissue, joint connective tissue, bone tissue; preferably, the central nervous tissue comprises spinal cord tissue, brain tissue; preferably, the peripheral nervous tissue comprises intra-articular nerve tissue, muscle nerve tissue; preferably, adipose tissue comprises subcutaneous adipose tissue, visceral adipose tissue.

56. The RNA inhibitor according to claim 55, wherein, The ocular tissue is retinal ganglion, endothelial cells, peripheral ocular muscle, or peripheral ocular fat.

57. The RNA inhibitor according to any one of claims 53 - 56, wherein, The delivery system is independently conjugated to one or more internal positions of the double-stranded ribonucleic acid.

58. The RNA inhibitor according to claim 57, wherein, The internal position is on a nucleobase, sugar ring, methylphosphonate bond, phosphorothioate bond, or phosphodiester bond.

59. The RNA inhibitor according to any one of claims 53 - 58, wherein, The delivery system is a lipophilic structure, and the lipophilic structure comprises a lipophilic group and a linker, and the lipophilic group is connected to the double-stranded ribonucleic acid through the linker.

60. The RNA inhibitor according to claim 59, wherein, The lipophilic structure is selected from aliphatic, alicyclic, and polyalicyclic compounds.

61. The RNA inhibitor according to claim 60, wherein, The lipophilic structure contains saturated or unsaturated C16 or C22.

62. The RNA inhibitor according to any one of claims 59 - 61, wherein, The linker is selected from a single bond, ether, thioether, urea, carbonate, amine, amide, maleimide-thioether, disulfide, phosphodiester, sulfonamide bond, product of click reaction, and carbamate.

63. The RNA inhibitor according to any one of claims 53-62, wherein, The delivery system is 64. The RNA inhibitor according to any one of claims 53 - 63, wherein the delivery system is conjugated to the 5'-end and / or 3'-end of the antisense strand or antisense nucleic acid fragment.

65. The RNA inhibitor according to any one of claims 53 - 64, wherein the delivery system is conjugated to the 5'-end and / or 3'-end of the sense strand or sense nucleic acid fragment.

66. The RNA inhibitor according to any one of claims 53 - 65, wherein the delivery system is conjugated to the 5'-end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 3'-end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

67. The RNA inhibitor according to any one of claims 53 - 66, wherein the delivery system is conjugated to the 3'-end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 5'-end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

68. The RNA inhibitor according to any one of claims 53 - 67, wherein the delivery system is conjugated to the 5'-end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 5'-end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

69. The RNA inhibitor according to any one of claims 53 - 68, wherein the delivery system is conjugated to the 3'-end of the antisense strand or antisense nucleic acid fragment, and the ligand is conjugated to the 3'-end of the sense strand or sense nucleic acid fragment, and the two ligands are the same or different.

70. The RNA inhibitor according to any one of claims 53 - 69, wherein the number of delivery systems is 1, 2, 3, 4, 5 or 6.

71. The RNA inhibitor according to any one of claims 53 - 70, wherein the antisense strand comprises at least 15 consecutive nucleotides in the sequence according to any one of SEQ ID NO: 822 - 827.

72. The RNA inhibitor according to any one of claims 53 - 71, wherein the sense strand comprises at least 15 consecutive nucleotides in the sequence according to any one of SEQ ID NO: 618 - 623.

73. The RNA inhibitor according to any one of claims 53 - 72, which is selected from: Z1, Z2, Z3, Z4, Z5, Z6.

74. A pharmaceutical composition, which comprises the RNA inhibitor according to any one of claims 1 - 73, and / or a physiologically acceptable excipient and / or carrier and / or diluent.

75. The composition according to claim 74, characterized in that The pharmaceutically acceptable carrier includes or is selected from aqueous carriers, liposomes, high molecular polymers or polypeptides.

76. Use of the RNA inhibitor targeting IGF-1R and its pharmaceutical composition in the preparation of a drug for treating IGF-1R-related diseases or pathologies; preferably, the RNA inhibitor is siRNA; more preferably, the siRNA is administered locally or to the lesion area tissue of a subject in need.

77. The use according to claim 76, wherein the IGF-1R-related diseases or pathologies include diseases or symptoms associated with elevated IGF-1R levels.

78. The use according to any one of claims 76 - 77, wherein the IGF-1R-related diseases or pathologies include thyroid eye disease, osteoarthritis, neuropathic pain.

79. The use according to any one of claims 76 - 78, wherein the RNA inhibitor and its pharmaceutical composition are the RNA inhibitor according to any one of claims 1 - 73 or the pharmaceutical composition according to any one of claims 74 - 75.

80. Use of the RNA inhibitor according to any one of claims 1-73 or the pharmaceutical composition according to any one of claims 74-75 in the manufacture of a medicament for preventing or treating a disease or pathology or reducing the risk of a disease or symptom.

81. Use according to claim 80, wherein the disease or pathology includes a disease or symptom associated with elevated IGF-1R levels.

82. Use according to any one of claims 80-81, wherein the disease or pathology includes thyroid eye disease, osteoarthritis, neuropathic pain.

83. A method for preventing or treating IGF-1R-related diseases or symptoms, comprising administering to a subject in need thereof an effective amount of an RNA inhibitor and its pharmaceutical composition.

84. Method according to claim 83, wherein the RNA inhibitor and its pharmaceutical composition are the RNA inhibitor according to any one of claims 1-73 or the pharmaceutical composition according to any one of claims 74-75.

85. Method according to claims 83-84, wherein the RNA inhibitor, its pharmaceutically acceptable salt or the pharmaceutical composition is administered to the subject by intraorbital injection, intra-articular injection, intrathecal injection, subcutaneous, intravenous, oral, rectal or intraperitoneal administration routes.

86. Method according to any one of claims 83-85, comprising administering to the local or lesion area tissue of a subject in need thereof.

87. Method according to claim 86, wherein the local or lesion area tissue of the subject in need thereof includes eye tissue, joint tissue, central nervous tissue, peripheral nerve tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

88. The method according to claim 87, wherein, The eye tissue is the optic nerve, trabecular meshwork, juxtacanalicular tissue, ganglion, episcleral vein, Schlemm's canal or peripheral eye tissue. Preferably, the joint tissue includes cartilage tissue, joint connective tissue, bone tissue; preferably, the central nervous tissue includes spinal cord tissue, brain tissue; preferably, the peripheral nerve tissue includes intra-articular nerve tissue, muscle nerve tissue; preferably, the adipose tissue includes subcutaneous adipose tissue, visceral adipose tissue.

89. The method according to claim 88, wherein, The eye tissue is retinal ganglion, endothelial cells, peripheral eye muscle or peripheral eye fat.

90. Method according to any one of claims 83-89, wherein the IGF-1R-related disease or pathology includes a disease or symptom associated with elevated IGF-1R levels.

91. Method according to any one of claims 83-90, wherein the IGF-1R-related disease or pathology includes thyroid eye disease, osteoarthritis, neuropathic pain.

92. A method for inhibiting IGF-1R gene expression in a cell, tissue or subject, comprising administering to the cell, tissue or subject an effective amount of the RNA inhibitor according to any one of claims 1-73 or the pharmaceutical composition according to any one of claims 74-75.

93. The method according to claim 92, wherein the disease or pathology comprises a disease or condition associated with elevated IGF-1R levels.

94. The method according to claim 93, wherein the disease or pathology comprises thyroid eye disease, osteoarthritis, neuropathic pain.

95. A method of preventing or treating a disease or condition, the method comprising administering to a subject in need thereof an effective amount of an RNA inhibitor according to any one of claims 1-73 or a pharmaceutical composition according to any one of claims 74-75.

96. The method according to claim 95, wherein the RNA inhibitor, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is administered to the subject by an intravitreal injection, intra-articular injection, intrathecal injection, subcutaneous, intravenous, oral, rectal, or intraperitoneal route of administration.

97. The method according to any one of claims 95-96, comprising administering to a local or lesion area tissue of a subject in need thereof.

98. The method according to claim 97, wherein the local or lesion area tissue of the subject in need thereof comprises ocular tissue, joint tissue, central nervous tissue, peripheral nerve tissue, tumor, liver tissue, kidney tissue, muscle tissue, or adipose tissue.

99. The method according to claim 98, wherein, The ocular tissue is the optic nerve, trabecular meshwork, juxtacanalicular tissue, ganglion, episcleral vein, Schlemm's canal, or peripheral ocular tissue. Preferably, the joint tissue comprises cartilage tissue, joint connective tissue, bone tissue; preferably, the central nervous tissue comprises spinal cord tissue, brain tissue; preferably, the peripheral nerve tissue comprises intra-articular nerve tissue, muscle nerve tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue, visceral adipose tissue.

100. The method according to claim 99, wherein, The ocular tissue is retinal ganglion, endothelial cells, peripheral ocular muscle, or peripheral ocular fat.

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