WO2025153084A1 - Rna inhibitor for inhibiting expression of insulin-like growth factor 1 receptor - Google Patents

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

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

Background Art

RNA interference (RNAi) technology refers to the phenomenon of highly conserved, highly efficient and specific degradation of homologous mRNA induced by double-stranded RNA (dsRNA) during evolution. It is a biological process in which RNA molecules inhibit the expression of a certain gene by destroying specific mRNA. Since RNAi technology can specifically eliminate or shut down the expression of specific genes, it has quickly become one of the most popular research tools in the field of gene function research and gene therapy research, and has been widely used to explore gene function, metabolic diseases, infectious diseases and the treatment of malignant tumors.

Thyroid eye disease (TED) is an autoimmune disease caused by hyperthyroidism. It is caused by the body's own immune system attacking the tissues around and behind the eyes due to the activation of the IGF-1R-mediated signaling complex on the cells in the orbit by self-antigens. The disease is characterized by the increase in the volume of the extraocular muscles, adipose tissue and connective tissue, and produces symptoms such as dry eyes, bulging eyes, retraction of eyelids, diplopia, strabismus, tingling with a sense of gravel, easy tearing, redness and swelling, and difficulty in eye movement. Severe cases can lead to changes in appearance and impaired vision.

IGF-1R (Insulin-like growth factor 1 receptor) is a tetrameric transmembrane receptor tyrosine kinase found on the surface of human cells and is the cell surface receptor for IGF-1 (hormone insulin-like growth factor 1). Studies have shown that IGF-1R is widely expressed in normal human tissues and highly expressed in thyroid eye disease (TED). Activation of IGF-1R can stimulate cell proliferation, survival, transformation, metastasis and angiogenesis, while inhibition of IGF-1R 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 by the US FDA for marketing in the United States. The drug is a fully human monoclonal antibody (mAb) and IGF-1R targeted inhibitor. Unlike other biologics such as anti-CD20 antibodies that can only reduce inflammation in the treatment of TAO, Tepezza can effectively reduce 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 had ended. However, Tepezza also had serious safety issues, with the most common adverse reactions including muscle spasms, hearing loss and hyperglycemia. These adverse reactions may be caused by systemic administration of the drug.

The development of locally administered IGF-1R siRNA is a solution to the current safety issues of monoclonal antibodies. Due to the long-term effect, siRNA-type drugs can bring a longer dosing cycle, thereby bringing better compliance. In addition, IGF-1R is also an important driving factor for tumor growth and development. The development of siRNA targeting IGF-1R also has a wide range of potential applications in the field of tumor treatment.

Summary of the invention

The present application provides an RNA inhibitor that inhibits the expression of the IGF-1R gene, providing an effective new therapy 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 a cell surface receptor for IGF-1. The RNA inhibitor of IGF-1R gene expression provided in 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 in 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 drug delivery efficiency through reasonable formulations.

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 chain, wherein the antisense chain 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.

In some embodiments, the antisense strand and the mRNA encoding IGF-1R (SEQ ID NO: 828) form a complementary region starting from any of the following positions counting from the 5' end:

Nucleotide No. 1118, Nucleotide No. 1120, Nucleotide No. 1154, Nucleotide No. 1367, Nucleotide No. 1407, Nucleotide No. 1415, Nucleotide No. 1532, Nucleotide No. 1625, Nucleotide No. 1627, Nucleotide No. 1628, Nucleotide No. 1631, Nucleotide No. 3356, Nucleotide No. 3357, Nucleotide No. 3359, Nucleotide No. 3792, Nucleotide No. 4198, Nucleotide No. 4200, Nucleotide No. 4208, Nucleotide No. 4685, Nucleotide No. 5246, Nucleotide No. 5392, Nucleotide No. 5393, Nucleotide No. 6329, Nucleotide No. 6332, Nucleotide No. 6333, Nucleotide No. 6334 nucleotide No. 4, 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. 53 nucleotide at position 80, nucleotide at position 5381, nucleotide at position 5663, nucleotide at position 5700, nucleotide at position 5711, nucleotide at position 5712, nucleotide at position 5721, nucleotide at position 5896, nucleotide at position 5898, nucleotide at position 6203, nucleotide at position 6236, nucleotide at position 6237, nucleotide at position 6240, nucleotide at position 6245, nucleotide at position 6288, nucleotide at position 6299, nucleotide at position 6305, nucleotide at position 6309, nucleotide at position 6310, nucleotide at position 6343, nucleotide at position 6363, nucleotide at position 6365, nucleotide at position 6366, nucleotide at position 6373, nucleotide at position 6379, nucleotide at position 6381 Acid, 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, nucleotide No. 8975, nucleotide No. 8977, nucleotide No. 9163, nucleotide No. 9 nucleotide No. 175, 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. 10266, nucleotide No. 10490, nucleotide No. 10526 , nucleotide No. 10528, nucleotide No. 10534, nucleotide No. 10535, nucleotide No. 10537, nucleotide No. 10540, nucleotide No. 10541, nucleotide No. 10679, nucleotide No. 10694, nucleotide No. 10717, nucleotide No. 10728, nucleotide No. 10729, nucleotide No. 10730, nucleotide No. 10754, nucleotide No. 10763, nucleotide No. 10766, nucleotide No. 10942, nucleotide No. 11034, nucleotide No. 11078, nucleotide No. 11126, nucleotide No. 11156, nucleotide No. 11159, nucleotide No. 11160, nucleotide No. 11292, Nucleotide at position 11340, nucleotide at position 11344, nucleotide at position 11383, nucleotide at position 11395, nucleotide at position 11789, nucleotide at position 11864, nucleotide at position 11865, nucleotide at position 11939, nucleotide at position 12092, nucleotide at position 12116, nucleotide at position 12130, nucleotide at position 12154, nucleotide at position 12156, nucleotide at position 12177, nucleotide at position 12195, nucleotide at position 1164, nucleotide at position 1163, nucleotide at position 1162, nucleotide at position 1161, nucleotide at position 1160, nucleotide at position 1165, nucleotide at position 1166, nucleotide at position 1167, and nucleotide at position 1168.

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

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

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

In some embodiments, it comprises a sense strand capable of forming a complementary double strand 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, and preferably the length of the duplex complementary region is 15-30 nucleotide pairs, more preferably 17-23 nucleotide pairs.

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, and more preferably the RNA inhibitor is siRNA.

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.

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.

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.

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

In some embodiments, 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.

In some embodiments, the antisense strand of the RNA comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:199-400.

In some embodiments, the RNA inhibitor, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:1-198.

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, ds-n35, ds-n36, ds-n37, ds-n3 8. 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-n 52. 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-n9 3. 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-n1 06. ds-n107, ds-n108, ds-n109, ds-n110, ds-n111, ds-n112, ds-n113, ds-n114, ds-n115, ds-n116, ds-n117, ds-n1 18. ds-n119, ds-n120, ds-n121, ds-n122, ds-n123, ds-n124, ds-n125, ds-n126, ds-n127, ds-n128, ds-n129, ds-n1 30. 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-n15 4. ds-n155, ds-n156, ds-n157, ds-n158, ds-n159, ds-n160, ds-n161, ds-n162, ds-n163, ds-n164, ds-n165, ds-n16 6. 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.

In some embodiments, the 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.

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.

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.

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.

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.

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.

In some embodiments, the inhibitory 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.

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.

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, respectively, the Z is G or modified G, and the N contains at least one nucleotide.

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

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

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

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.

In some embodiments, the RNA inhibitor, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:401-413.

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.

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

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.

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-methylureidine (m5U) modification, pseudouridine modification, 2-thioureidine (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.

In some embodiments, in the RNA inhibitor, 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.

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).

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

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.

In some embodiments, in the RNA inhibitor, 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.

In some embodiments, the RNA inhibitor has the following modified combinations:

a) the positive 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;

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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides.

In some embodiments, the RNA inhibitor has the following modified combinations:

a) the positive strand comprises a first sequence of 16-21 nt in length, wherein the first sequence comprises the following modifications: counting from the 5' end, the 9th, 10th and 11th nucleotides are 2'-F modified, and the rest are 2'-Ome modified;

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 connection between the 1st and 2nd nucleotides, there is a phosphorothioate connection between the 2nd and 3rd nucleotides, and there is a phosphorothioate connection between the 3rd and 4th nucleotides; and counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides; and counting from the 5' end, there is a 2'-F modification at the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides, and the rest are 2'-Ome modifications.

In some embodiments, the RNA inhibitor has the following modified combinations:

a) the positive 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;

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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides.

In some embodiments, the RNA inhibitor, wherein the first sequence length is preferably 16nt, 17nt, 18nt, 19nt, 20nt, 21nt, preferably 21nt.

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

In some embodiments, the RNA inhibitor has the following modified combinations:

a) The forward strand is 21 nt long and includes the following modifications: Counting from the 5' end, nucleotides at positions 9, 10, and 11 have 2'-F modifications;

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 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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides.

In some embodiments, the RNA inhibitor has the following modified combinations:

a) The forward strand is 21 nt long and 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;

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 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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides, and the rest are 2'-Ome modifications.

In some embodiments, the RNA inhibitor, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:624-812.

In some embodiments, the RNA inhibitor, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:414-602.

In some embodiments, the RNA inhibitor 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-m1 4. 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, d s-m113, ds-m114, ds-m115, ds-m116, ds-m117, ds-m118, ds-m119, ds-m120, ds-m121, ds-m122, ds-m123, d s-m124, ds-m125, ds-m126, ds-m127, ds-m128, ds-m129, ds-m130, ds-m131, ds-m132, ds-m133, ds-m134, d s-m135, ds-m136, ds-m137, ds-m138, ds-m139, ds-m140, ds-m141, ds-m142, ds-m143, ds-m144, ds-m145, d s-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.

In some embodiments, the RNA inhibitor, wherein

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 connection between the 1st and 2nd nucleotides of the sense strand, there is a phosphorothioate connection between the 2nd and 3rd nucleotides, there is a 2'-F modification at the 9th, 10th, 11th, and 18th nucleotides of the sense strand, and the rest are 2'-OMe modified; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides;

b) the antisense strand comprises a fourth sequence with a length of 19-26 nt, wherein the fourth sequence includes the following modifications: counting from the 5' end, the nucleotides at positions 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 connection between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides.

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

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

In some embodiments, any one of the RNA inhibitors, wherein

a) The length of the sense strand is 21 nt. Counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides. The 9th, 10th, 11th, and 18th nucleotides 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 the 1st and 2nd nucleotides, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides;

b) The antisense chain is 26 nt in length. Counting from the 5' end, the nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense chain are modified with 2'-F, and the rest are modified with 2'-OMe. 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.

In some embodiments, the RNA inhibitor, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:813-827.

In some embodiments, the RNA inhibitor, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:603-617.

In some embodiments, the RNA inhibitor comprises a duplex selected from the following:

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.

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

In some embodiments, the target tissue is eye tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, adipose tissue.

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.

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

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

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

In some embodiments, the delivery system is a lipophilic structure, 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.

In some embodiments, the lipophilic structure is selected from aliphatic, alicyclic and polyaliphatic compounds.

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

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.

In some embodiments, the delivery system is

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.

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.

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.

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.

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.

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.

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

In some embodiments, the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:822-827.

In some embodiments, the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:618-623.

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

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.

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.

On the other hand, the present application provides an RNA inhibitor targeting IGF-1R and a pharmaceutical composition thereof 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.

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

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

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

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

In another aspect, the present application provides 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.

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

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.

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

In some embodiments, the local or focal 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.

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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

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

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

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

Those skilled in the art can easily discern other aspects and advantages of the present application from the detailed description below. In the detailed description below, only exemplary embodiments of the present application are shown and described. As will be appreciated by those skilled in the art, the content of the present 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 the present application relates.

BRIEF DESCRIPTION OF THE DRAWINGS

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

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

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

The following is an explanation of the implementation of the present invention by means of specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

Definition of terms

In the present application, "optional", "optional" or "optionally" are equivalent in meaning, meaning that the event or situation described thereafter may or may not occur, and the description includes situations where the event or situation occurs and situations 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 containing one or more substituents, these groups are not intended to introduce any substitution or substitution pattern that is sterically impractical, synthetically infeasible and/or inherently unstable. For example, "optionally modified" includes not being modified and being modified, and further, "nucleotides are optionally modified" includes nucleotides that are not modified and nucleotides that are modified.

In this application, when any variable (e.g., a substituent R, such as a nucleic acid being modified) appears 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 by 0-2 Rs, the group may be optionally substituted by up to two Rs, and each case has independent options. For another example, when multiple nucleotides are modified, each nucleotide is optionally modified independently, and the type and amount of modification of each nucleotide may be the same or different.

In this application, the terms "RNA inhibitor", "iRNA", "siRNA", "RNAi agent", "iRNA agent" and "RNA interfering agent" are used interchangeably and generally refer to an agent comprising RNA as defined in the terms of this application, which can mediate targeted cleavage of RNA transcripts by forming an RNA-induced silencing complex (RISC) in cells. The RNA inhibitor directs the sequence-specific degradation of mRNA via a process called RNA interference (RNAi). The RNA inhibitor described in the present application can regulate or inhibit gene expression of the IGF-1R gene in cells, and in some embodiments, the cells can be cells of mammalian subjects. 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), and the mRNA sequence includes the sequence shown in SEQ ID NO:828.

In certain embodiments, the "RNA inhibitor" used in the present 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 cleaves 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 method described in Lima et al. (2012) Cell 150: 883-894.

In certain embodiments, the "RNA inhibitor" used in the present 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 strand and/or antisense strand of the "siRNA duplex" in a stem-loop or hairpin structure. The length of the stem of this hairpin structure is generally in the range of 19 to 29 nucleotides, and the length of the loop is generally 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).

In certain embodiments, the "RNA inhibitor" used in the present application is a miRNA. The term "miRNA" or "microRNA" is used herein according to its common meaning 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. Mature fully processed miRNAs are about 15 to about 30 nucleotides in length. A representative set of known endogenous miRNA species are 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 at the Wellcome Trust Sanger Institute website. The mature fully processed miRNAs publicly available on the miRBase sequence database are incorporated herein by reference. A representative set of miRNAs is also included in Table 1 below this article. 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.

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 binds to a target RNA through RNA-RNA or RNA-DNA or RNA-PNA (protein nucleic acid; Egholm et al., 1993 Nature 365, 566) interactions, and alters the activity of the target RNA through spatial interactions or through RNase H-mediated target recognition (for review, see Stein and Cheng, 1993 Science 261, 1004 and Woolf et al., U.S. Patent No. 5,849,902). Generally, antisense molecules are complementary to target sequences along a single adjacent sequence of the antisense molecule. However, in certain embodiments, the antisense molecule can bind to a substrate so that the substrate molecule forms a ring, and/or the antisense molecule can bind so that the antisense molecule forms a ring. Thus, an antisense molecule can be complementary to two (or even more) non-adjacent substrate sequences, or two (or even more) non-adjacent 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 N. A. Drug 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 through DNA-RNA interactions, 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 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.

In certain embodiments, the "RNA inhibitor" used in the present application is a 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 chains, which have two chains in "sense" and "antisense" orientations relative to the target RNA (i.e., the IGF-1R gene). In some embodiments, the siRNA duplex directs the sequence-specific degradation of mRNA (such as the mRNA sequence of the IGF-1R gene) via a process called RNA interference (RNAi).

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

The duplex structure can be any length that allows for specific degradation of the desired target RNA by the RISC pathway, and can be within the length range of about 19 to 36 base pairs, 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 the present application. In certain embodiments, the RNA inhibitor of the present application is a dsRNA having 15-23 nucleotides in each chain, which interacts with a target RNA sequence (e.g., an IGF-1R gene) to guide the cutting of the target RNA. In certain embodiments, the RNA inhibitor of the present application is a dsRNA of 24-30 nucleotides, which interacts with a target RNA sequence (e.g., an mRNA sequence of an IGF-1R gene) to guide the cutting of the target RNA.

In the present application, the term "antisense strand" generally refers to a strand on 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 mRNA precursors and mRNA molecules, for example, an mRNA sequence of an IGF-1R gene). The term "complementarity region" used in the present application generally refers to a region on the antisense strand that is substantially complementary to the (mRNA sequence of an IGF-1R gene) used in the present application. When the complementary region is not completely complementary, the mismatch may be in the interior or terminal regions of the molecule. Generally, the most tolerated mismatch is in the terminal region, for example, within 5, 4, 3, or 2 nucleotides of the 5' end and/or 3' end.

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.

In the present application, the term "antisense nucleic acid fragment" refers to a continuous fragment on the antisense strand, and the fragment length may include 15-35 nucleotides. For example, the antisense nucleic acid fragment may 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 3 continuous nucleotide fragments on the antisense nucleic acid fragment are complementary to continuous nucleotide fragments of corresponding length in the mRNA encoding IGF-1R.

In the present application, the term "sense strand" generally refers to a strand of an RNA inhibitor, comprising a region substantially complementary to the region of the term antisense strand as defined herein. A "sense" strand is sometimes referred to as a "sense" strand, a "passenger" strand or an "anti-guide" strand. With 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.

In the present application, the term "sense nucleic acid fragment" refers to a continuous fragment on the sense strand, and the fragment length may include 15-35 nucleotides. For example, the sense nucleic acid fragment may 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.

In the present 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.

In the present application, the term "fully complementary" generally means that all (100%) bases in the continuous sequence of the antisense strand or antisense nucleic acid fragment of the RNA inhibitor will hybridize with the same number of bases in the continuous sequence of the sense strand or sense nucleic acid fragment of the RNA inhibitor or IGF-1R mRNA. The continuous sequence may contain all or part of the above sequence. As used in the present application, "partially complementary" generally means that in the hybridized nucleobase sequence pair, at least about 70% of the bases in the continuous sequence of the antisense strand or sense nucleic acid fragment of the RNA inhibitor will hybridize with the same number of bases in the continuous sequence of the sense strand or sense nucleic acid fragment of the RNA inhibitor or IGF-1R mRNA. The terms "complementary", "fully complementary" and "substantially complementary" as used in the present application can be used in terms of base matching between the sense strand or sense nucleic acid fragment of the RNA inhibitor and the antisense strand or antisense nucleic acid fragment or between the antisense strand or antisense nucleic acid fragment of the RNA inhibitor and the sequence of the 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.

As used herein, the term "nucleotide" refers to a pentose (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 some embodiments, the nucleotide is an unmodified ribonucleotide. In some embodiments, the ribonucleotide is a 3'-ribonucleotide. In some embodiments, the ribonucleotide is a 5'-ribonucleotide. In some embodiments, modified or unmodified nucleotides may be optionally further modified.

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.

The natural bases of RNA include A (adenine), G (guanine), C (cytosine), U (uracil) and T (thymine). As used herein, the marking "d" before the monomer (such as nucleotides A, U, C, G and T, etc.) indicates that the monomer is modified by 2'-deoxy. As used herein, the marking "f" after the monomer (such as nucleotides A, U, C, G and T, etc.) indicates that the monomer is modified by 2'-deoxy-2'-fluorine (2'-F modification). As used herein, the marking "GNA-" before the monomer (such as nucleotides A, U, C, G and T, etc.) indicates that the monomer is modified by ethylene glycol nucleic acid (GNA modification). As used herein, Tgn is the code for GNA-T, which is equivalent to the meaning. As used herein, 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 by 2'-O-methyl (2'-OMe).

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:

In the present invention, the mark "d" before a nucleotide (A, U, C, G, T, etc.) indicates that the nucleotide is 2'-deoxy modified. Exemplarily, the structure of a 2'-deoxy modified nucleotide is as follows:

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

In the present invention, "GNA-" is marked before a nucleotide (A, U, C, G, T, etc.), indicating that the nucleotide is modified by ethylene glycol nucleic acid (GNA modification). In the present invention, Tgn is the code for GNA-T, which is equivalent. Exemplarily, the structure of the nucleotide modified by GNA is as follows:

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 by 2'-O-methyl (2'-OMe). Exemplarily, the structure of the nucleotide modified by 2'-OMe is as follows:

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

For example, The structure modified by invAB:

In the present invention, invAb modification refers to the modification of replacing a nucleotide with an inverted abasic nucleotide (invAb). Exemplary, The structure after modification by invAb:

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

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., phosphorothioate diester bond), i.e., modified by phosphorothioate (PS).

In the present invention, the absence of a "*" 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).

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 via a phosphorothioate bond.

As used herein, "optional", "optional" or "optionally" are equivalent in meaning, meaning that the event or situation described thereafter may or may not occur, and the description includes both situations where the event or situation occurs and situations 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 containing one or more substituents, these groups are not intended to introduce any substitution or substitution pattern that is sterically impractical, synthetically infeasible and/or inherently unstable. For example, "optionally modified" includes not being modified and being modified, and further, "nucleotides are optionally modified" includes nucleotides that are not modified and nucleotides that are modified.

As used herein, when any variable (e.g., a substituent R, such as a nucleic acid that is modified) appears 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 be optionally substituted with up to two Rs, and each case of R has independent options. For another example, when multiple nucleotides are modified, each nucleotide is optionally modified independently, and the type and amount of modification of each nucleotide can be the same or different.

As used herein, unless otherwise specified, "comprising," "including," "at least," "having," "having," "at least," "containing," 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.

As used herein, "complementary" or "reverse complementary" can be 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., a purine nucleotide of a nucleic acid that is complementary to a pyrimidine nucleotide of an 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) is always paired with thymine (T), and in RNA, adenine (A) is paired with uracil (U); guanine (G) is always paired 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, nucleotides containing hypoxanthine as its base may be base paired with nucleotides containing adenine, cytosine or uracil. In some embodiments of the present application, nucleotides containing uracil, guanine or adenine may be replaced in the nucleotide sequence of the present application by nucleotides containing, for example, inosine (in the application, the capital letter "I" may represent hypoxanthine base, inosine or nucleotides 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 G-U wobble base pairing with the target mRNA.

The degree of complementarity of an oligonucleotide to another oligonucleotide is called complementarity, which is measured by the percentage of bases that can form hydrogen bonds with each other in each chain, which is determined by established base pairing rules. Oligonucleotide sequences do not need to be "completely complementary" (i.e., "perfectly complementary") to their corresponding nucleic acid sequences. In some embodiments, if a first nucleotide sequence and a second nucleotide sequence show at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% degree of sequence complementarity, then the first nucleotide sequence can be considered to be complementary to 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 to achieve 90% complementarity. Non-complementary core bases are also referred to as "mismatches" and may be gathered or scattered between complementary bases, and do not need to be adjacent to each other or adjacent to complementary core bases.

As used herein, "mismatches" include, but are not limited to:

1) Two opposite (independent natural or unnatural) nucleotides (other than A-T, A-U or G-C) pair;

2) No hydrogen bonds are formed between two opposing (independent natural or non-natural) nucleotides;

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

In some embodiments, the mismatches include wobble base pairing and Hoogstein base pairing.

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 a fully complementary region. A "fully complementary" oligonucleotide may include an internal region (e.g., at least 7, 8, 9, or 10 nucleotides) that is fully complementary to the target RNA.

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 degradation metabolism. In some embodiments, the blocking group can reduce or inhibit the RNA interference effect of the oligonucleotide. In some 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, M03 and M06.

As used herein, double-stranded nucleotide reagents can be optionally conjugated with one or more blocking groups. The blocking group can be connected to the sense strand, antisense strand or two strands at the 3' end, 5' end or both ends. In certain embodiments, the blocking group is conjugated to the antisense strand, specifically to the 5' end of the antisense strand. In certain embodiments, the blocking group is conjugated to an oligonucleotide (e.g., the 5' end of the antisense strand) by a nucleotide bond, and the internucleotide bond is optionally modified as described herein. In certain embodiments, the blocking group is connected to the double-stranded nucleotide reagent by a thiophosphate. In certain embodiments, the blocking group is connected to the double-stranded nucleotide reagent by a phosphodiester bond.

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

In this application, the terms "nucleic acid" and "polynucleotide" are used interchangeably and refer to a polymeric form of nucleotides (deoxyribonucleotides or ribonucleotides or analogs thereof) 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 present application as a polynucleotide 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, a primer, an EST or SAGE tag), an exon, an intron, a messenger RNA (mRNA), a transfer RNA, a ribosomal RNA, a ribozyme, a cDNA, a recombinant polynucleotide, a branched polynucleotide, a plasmid, a vector, an isolated DNA of any sequence, an isolated RNA of any sequence, a nucleic acid probe, siRNA, miRNA, shRNA, an RNAi agent, and a primer.

In the present application, the term "oligonucleotide" generally refers to a polymer composed of a plurality of nucleotide residues (deoxyribonucleotides or ribonucleotides, or its related structural variants or synthetic analogs) connected by a phosphodiester bond (or its related 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 acid (PNA), phosphoramidate, phosphorothioate, methylphosphonate, 2-O-methylribonucleic acid, etc. The exact size of the molecule can depend on specific applications. Oligonucleotides are generally shorter in length, usually approximately having 10-30 nucleotide residues, but the term can also refer to molecules of any length, although the terms "polynucleotide" or "nucleic acid" are generally used for larger oligonucleotides.

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.

In the present application, the term "modified nucleoside" generally means a nucleoside comprising at least one chemical modification compared to a naturally occurring RNA or DNA nucleoside. The modified nucleoside comprises a modified sugar moiety and/or a modified nucleobase.

In the present 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 substitutes for moieties. As used in the present application, "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 naturally occurring heterocyclic nucleobases of RNA or DNA: purine bases adenine (a) and guanine (g); and thymine (t), cytosine (c) (including 5-methyl c) and uracil (u). "Modified nucleobase" generally refers to any nucleobase that is not a naturally occurring nucleobase.

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

In this application, the term "internucleoside linkage" generally means a covalent linkage between adjacent nucleosides in an oligonucleotide. A "naturally occurring internucleoside linkage" means a 3' to 5' phosphodiester linkage. A "modified internucleoside linkage" means any internucleoside linkage other than a naturally occurring internucleoside linkage.

In this application, the term "target nucleic acid" or "target sequence" generally refers to a continuous portion of the nucleotide sequence of an mRNA molecule formed during transcription of the IGF-1R gene, including mRNA that is an RNA processing product of the primary transcription product. The target portion of the sequence should be at least long enough to serve as a substrate for iRNA-guided 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 about 19-36 nucleotides in length, for example, preferably about 19-30 nucleotides in length. Ranges and lengths intermediate to the above ranges and lengths are also included as part of this application.

In the present application, the term "IGF-1R protein" refers to a human tetrameric transmembrane tyrosine kinase, which 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. In the present application, the term "IGF-1R gene" refers to a gene encoding an IGF-1R protein, which has a molecular weight of about 320 kDa and is composed of two α subunits and two β subunits. Among them, the IGF-1R gene can be regulated or inhibited by an RNA inhibitor, and 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). Among them, the transcription sequence of the IGF-1R gene is as shown in SEQ ID NO: 828.

In the present application, the term "blocking group" refers to a group that can be selectively combined with a duplex siRNA described in the present application or a drug comprising a duplex siRNA, and the blocking group is located on a sense strand or a sense nucleic acid fragment, an antisense strand or an antisense nucleic acid fragment or two chains. The blocking group can be attached to a sense strand or a sense nucleic acid fragment, an antisense strand or an antisense nucleic acid fragment or 3' end, 5' end or two ends of the two chains. In certain embodiments, the blocking group is attached to the antisense strand or the antisense nucleic acid fragment, particularly the 5' end of the antisense strand or the antisense nucleic acid fragment. In certain embodiments, the blocking group is linked to an oligonucleotide (for example, the 5' end of the antisense strand or the antisense nucleic acid fragment) by an internucleotide link, and the internucleotide link is optionally modified as described above. In certain embodiments, the blocking group is linked to a double-stranded oligonucleotide agent by phosphosulfate. In certain embodiments, the blocking group is linked to a double-stranded oligonucleotide agent by a phosphodiester. In the present application, in order to reduce or inhibit the degradation of exonucleases, a "blocking group" may be attached to the 5' end of the duplex siRNA antisense strand or antisense nucleic acid fragment described in the present application. 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).

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, and the receptor that interacts with the ligand can be present on the cell surface, or alternatively can be an intracellular and/or intercellular receptor, and the interaction of the ligand with the receptor can result in a biochemical reaction, or can be simply a physical interaction or combination.

In the present application, "conjugation" refers to the connection between two or more chemical parts each having a specific function in a covalently linked manner; accordingly, "conjugate" refers to a compound formed by covalently linking the chemical parts. For example, "double-stranded RNA conjugate" means a compound or complex formed by covalently linking one or more chemical parts (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 covalently linked to the atom at the 3' or 5' position of the nucleotide through a phosphodiester bond), 2'-hydroxyl, 5'-hydroxyl 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, and the nucleotides are connected by a 2'-5' phosphodiester bond. In some embodiments, the delivery system, ligand group or conjugated group can also be connected to the 3'-position of the nucleotide, and the nucleotides are connected by a 3'-5' phosphodiester bond. In some embodiments, the connection between the delivery system, ligand group or conjugated group and the nucleotide can be further modified, such as: thio modification (i.e., connected by a thiophosphodiester bond).

In this application, the terms "induce", "inhibit", "enhance", "elevate", "increase", "reduce", "lower" 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 activity or gene expression of IGF-1R in the treated sample will be lower than the level of IGF-1R activity or gene expression in the untreated sample. The 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. "Reduce" and "reduce" are relative terms and require comparison between before and after measurement. "Reduce" and "reduce" include complete depletion.

In certain embodiments, the term "reduction" can be detected by standard methods known in the art (such as those described in the present application), and the gene expression level/amount of a gene, gene product, such as a protein or a biomarker in a first sample is compared with the gene expression level/amount of a corresponding gene, gene product, such as a protein or a biomarker in a second sample by about 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 100% overall reduction. In certain embodiments, the term "reduction" refers to the reduction of 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.

In this application, the term "gene expression" generally refers to the process by which a gene ultimately produces a protein. The gene expression includes, but is not limited to, transcription, post-transcriptional modification (e.g., splicing, polyadenylation, addition of a 5'-cap) and translation.

In the present 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 preparations may generally contain salts, excipients, buffers, preservatives, compatible carriers and optional other therapeutic agents. Such pharmaceutically acceptable preparations may also generally contain 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 they cannot be 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, etc. Pharmaceutically acceptable salts can also be prepared as alkali metal salts or alkaline earth metal salts, such as sodium salts, potassium salts or calcium salts.

In the present application, the term "prevention and/or treatment" includes not only prevention and/or treatment of diseases, but also generally includes prevention of the onset of diseases, slowing or reversing the progression of diseases, preventing or slowing the onset of one or more symptoms associated with diseases, reducing and/or alleviating one or more symptoms associated with diseases, reducing the severity and/or duration of diseases and/or any symptoms associated therewith, and/or preventing further increase in the severity of diseases and/or any symptoms associated therewith, preventing, reducing or reversing any physiological damage caused by diseases, and any pharmacological effects that are generally beneficial to patients being treated. The RNAi agent or pharmaceutical composition of the present application does not need to achieve complete cure or eradication of any symptoms or manifestations of diseases 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, prophylactically administered treatments constitute viable preventive agents that do not need to completely and effectively prevent the onset of symptoms. It is sufficient to simply reduce 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 reduce the likelihood of disease occurrence or exacerbation.

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

In the present application, the term "administering" generally refers to introducing the present application's 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.

In the present application, the term "contact" generally refers to two or more different types of substances being in contact with each other 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 direct contact of the RNAi agent or composition of the present application with a cell or tissue. In other embodiments, the term refers to indirect contact of the RNA inhibitor or composition of the present application with a cell or tissue. 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 a cell or tissue by diffusion or any other active transport or passive transport process known in the art (by which the compound circulates in the body).

In the present 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 a drug that promotes disease regression (which is demonstrated by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease asymptomatic periods, or the prevention of damage or disability caused by suffering from a disease) when used alone or in combination with another therapeutic agent. A "preventive effective amount" or "preventive effective dose" of a drug generally refers to an amount of a drug that inhibits the development or recurrence of a disease when administered alone or in combination with another therapeutic agent to a subject at risk of disease development or disease recurrence. The ability of a therapeutic agent or preventive agent to promote disease regression or inhibit disease development or recurrence can be evaluated using a variety of methods known to those skilled in the art, such as in human subjects during clinical trials, in animal model systems predicting efficacy in humans, or by measuring the activity of a pharmaceutical agent in an in vitro assay. In certain embodiments, an "effective amount" refers to the amount of an RNA inhibitor that produces an expected pharmacological, therapeutic or preventive result.

In this application, the term "subject" generally refers to a human or non-human animal (including mammals) that needs to diagnose, prognose, improve, prevent and/or treat 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.

In this application, the terms "include", "comprising", "having", "may", "containing" and their variations 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 means that no other components (or similarly, features, integers, steps, etc.) can be present. Unless the context clearly dictates otherwise, singular forms such as "a", "an", "the" in English, "a", "a kind" and "described/the" in Chinese generally include plural forms of the referred things.

In this application, the term "about" generally means roughly, roughly, or around. When the term "about" is used to refer to a numerical range, a cutoff or a specific value is used to indicate that the stated value may differ from the recited value by up to 10%. Thus, the term "about" can be used to cover variations of ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from a specific value.

It should be understood that the term "at least" preceding a number or a series of numbers includes the numbers adjacent to the term "at least", and all subsequent numbers or integers 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 nucleic acid molecule of 21 nucleotides" means that 19, 20 or 21 nucleotides have the indicated properties. When "at least" appears before a series of numbers or ranges, it should be understood that "at least" can modify each number in the series or range.

It should be understood that "no more than" or "less than" as used herein refers to a value or integer that is adjacent to the phrase and logically lower, such as from the logic of the context, to zero. For example, a duplex having an overhang of "no more than 3 nucleotides" has an overhang of 3, 2, 1 or 0 nucleotides. When "no more than" appears before a series of numbers or ranges, it should be understood that "no more than" can modify each number in the series or range. As used herein, ranges include both upper and lower limits.

DETAILED DESCRIPTION OF THE INVENTION

Antisense strand or antisense nucleic acid fragment and sense strand or sense nucleic acid fragment

On the one hand, the present application provides an RNA inhibitor for inhibiting the expression of 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 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 length of the duplex complementary region is 15-30 nucleotide pairs, more preferably 17-23 nucleotide pairs. For example, the length of 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 certain embodiments, the RNA inhibitor comprises a single-stranded oligonucleotide or double-stranded ribonucleic acid (dsRNA) molecule for inhibiting the expression of the IGF-1R gene in a cell, such as a cell of a subject (e.g., a mammal). The dsRNA comprises an antisense strand or an antisense nucleic acid fragment having a complementary region complementary to at least a portion of the mRNA formed in the expression of the IGF-1R gene. 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).

dsRNA includes 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 (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 (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 suitable conditions, the two chains can hybridize and form a duplex structure. Generally, 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.

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 cutting 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 cutting (i.e., cutting via the RISC pathway).

It will also be understood by those skilled in the art that the complementary region is the major functional portion of the dsRNA, e.g., a 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 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs.

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.

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.

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.

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.

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.

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.

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

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.

In some embodiments, the antisense strand or antisense nucleic acid fragment is selected from any one of SEQ ID NO: 199-413 or at least 15 consecutive nucleotide sequences that differ from it by no more than 3 nucleotides, for example, 15, 16, 17, 18, 19 or 20 consecutive nucleotides.

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

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

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 3' or 5' overhangs of 1-3 nucleotides in length, or the antisense strand has 3' or 5' overhangs of 1-3 nucleotides in length. In some embodiments, the RNA inhibitor described in the present application has a specific structure near its 5' end and 3' end, as shown in Figure 2.

In certain embodiments, the antisense strand comprises a cleavage region, and the cleavage region comprises a nucleotide sequence represented by formula (I),

Formula (I): (3'-5')X2-Y-Z,

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 a guanine nucleotide (G), a natural analog of a guanine nucleotide (G), a non-natural analog of a guanine nucleotide (G), an adenine nucleotide (A), a natural analog of adenine nucleotide (A), or a non-natural analog of adenine nucleotide (A).

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

In certain embodiments, X2 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).

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.

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).

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.

In certain embodiments, the cleavage region further comprises a nucleotide N1, wherein N1 is the third nucleotide from the 3' end of the 5' extension, and the cleavage region comprises a nucleotide sequence represented by formula (II),

Formula (II): (3’-5’)X2-Y-Z-N1.

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 the nucleotide sequence shown in formula (III),

Formula (III): (3’-5’)X2-Y-Z-N,

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

In certain embodiments, N is an adenine nucleotide (A), a guanine nucleotide (G), a cytosine nucleotide (C), a uracil nucleotide (U), their natural analogs or their non-natural analogs; preferably, N1 is a cytosine nucleotide (C), a natural analog of a cytosine nucleotide (C) or a non-natural analog of a cytosine nucleotide (C).

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

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 in which each sequence in Table 1 differs by one, two or three nucleotides, for example, 15, 16, 17, 18, 19 or 20 consecutive nucleotides.

Table 1 siRNA sense and antisense strand sequences

Modified nucleotides

In order to enhance the stability of the RNA inhibitor in vivo, the sense strand and antisense strand of the RNA inhibitor may be modified without affecting its activity or even enhancing its activity, wherein the nucleotides may have a modifying group, and the entire strand or a portion thereof may be modified. In certain embodiments, one or more nucleotides on the sense strand and/or antisense strand are modified to form modified nucleotides.

As used herein, the term "nucleotide" refers to a pentose (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 some embodiments, the nucleotide is an unmodified ribonucleotide. In some embodiments, the ribonucleotide is a 3'-ribonucleotide. In some embodiments, the ribonucleotide is a 5'-ribonucleotide. In some embodiments, modified or unmodified nucleotides may be optionally further modified.

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.

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 nucleotides A, U, C, G and T, etc.) indicates that the monomer is modified by 2'-deoxy.

As used herein, the label "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).

As used herein, "GNA-" is marked before a monomer (such as nucleotides A, U, C, G and T, etc.), indicating 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 in meaning.

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).

As used herein, the invAB modification refers to the bonding of an inverted abasic nucleotide to the 5' or 3' end of the nucleotide. The structure modified by invAB:

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

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).

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, phosphodiester bond).

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

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:

(1) modifying the phosphodiester bonds of nucleotides in the nucleotide sequence of the small inhibitory nucleic acid molecule;

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

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

The chemical modification of the present invention is well known to those skilled in the art, and the modification of the phosphodiester bond refers to the modification of the oxygen in the phosphodiester bond, including phosphorothioate modification and borylated phosphate modification. Both modifications can stabilize the siRNA structure and maintain high specificity and high affinity of base pairing.

The ribose modification refers to the modification of the 2'-OH in the pentose of the nucleotide, that is, introducing 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.

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.

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

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

The term "LNA" refers to a bicyclic nucleoside analogue comprising a C2*-C4* diradical (bridge), and is referred to as a "locked nucleic acid". It may refer to an LNA monomer, or, when used in the context of an "LNA oligonucleotide", LNA refers to an oligonucleotide containing one or more such bicyclic nucleotide analogues. In certain aspects, a bicyclic nucleoside analogue is an LNA nucleotide, and these terms may therefore be used interchangeably, and in such embodiments, both are characterized by the presence of a linking group (such as a bridge) between C2' and C4' of the ribose sugar ring.

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

Where B is a base.

GNA (glycerol nucleic acid) is a chemical substance similar to DNA or RNA, but with a different composition and does not exist in any currently known natural organism. GNA contains an acyclic three-carbon propylene glycol (1,2-propanediol) backbone, which replaces the (deoxy)ribose of DNA and RNA to form the simplest chemically stable nucleic acid structure. The S-(GNA) structure is shown below.

Where B is a base.

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

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

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), 5-methylcytidine (m5C), 3-methyluridine (m3U), 5-methylureidine (m5U), pseudouridine, 2-thioureidine (s2U), propyne uridine (5-pU), inverted abasic nucleotide (invAB) modification by bonding the 5' or 3' end of the nucleotide, inverted abasic nucleotide (invAb) modification, nucleotide replacement with an inverted abasic nucleotide (rF) modification, nucleotide replacement with a 2,4-difluoromethylphenylribonucleotide (rF) modification, or nucleotide replacement 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.

In certain embodiments, in the RNA inhibitor, when the length of the sense strand is 21 nt, counting from the 5' end, the 9th nucleotide of the sense strand is modified with 2'-F.

In certain embodiments, in the RNA inhibitor, when the length of the sense strand is 21 nt, counting from the 5' end, the 10th nucleotide of the sense strand is modified with 2'-F.

In certain embodiments, in the RNA inhibitor, when the length of the sense strand is 21 nt, counting from the 5' end, the 11th nucleotide of the sense strand is modified with 2'-F.

In certain embodiments, the RNA inhibitor, when the length of the sense strand is 21 nt, 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.

In certain embodiments, the RNA inhibitor, when the length of the positive strand is 21 nt, counting starts from the 5' end, wherein the 5' end of the first nucleotide of the positive strand has an invAB modification.

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.

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 1st and 2nd nucleotides of the antisense strand.

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.

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.

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 1st and 2nd nucleotides of the antisense strand.

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

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 second nucleotide of the antisense strand.

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 3rd nucleotide of the antisense strand.

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.

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.

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.

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.

In certain embodiments, the RNA inhibitor, when the antisense strand 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.

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

In certain embodiments, the RNA inhibitor,

a) When the length of the sense strand is 21 nt, the sense strand includes the following modifications: counting from the 5' end, the 9th, 10th, and 11th nucleotides are modified with 2'-F, and the rest are modified with 2'-OMe; counting from the 5' end, the 5' end of the 1st nucleotide is modified with invAB,

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

In certain embodiments, the RNA inhibitor, when the length of the positive strand is 21 nt, counting from the 5' end, the 18th nucleotide has a 2'-F modification.

In certain embodiments, the RNA inhibitor, when the length of the positive strand is 21 nt, 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.

In certain embodiments, the RNA inhibitor, when the length of the positive strand is 21 nt, counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides.

In certain embodiments, the RNA inhibitor, when the length of the positive strand is 21 nt, counting from the 5' end, there is a phosphorothioate linkage between the 2nd and 3rd nucleotides.

In certain embodiments, the RNA inhibitor, when the length of the positive strand is 21 nt, counting from the 3' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides.

In certain embodiments, the RNA inhibitor, when the length of the positive strand is 21 nt, counting from the 3' end, there is a phosphorothioate linkage between the 2nd and 3rd nucleotides.

In certain embodiments, the RNA inhibitor, when the length of the sense chain is 21 nt, counting from the 5' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides of the sense chain, there is a phosphorothioate connection between the 2nd and 3rd nucleotides, the 9th, 10th, 11th, and 18th nucleotides of the sense chain are 2'-F modified, and the rest are 2'-OMe modified; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides of the sense chain, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides of the sense chain.

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.

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 second nucleotide of the antisense strand.

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.

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.

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.

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.

In certain embodiments, the RNA inhibitor, 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.

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

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

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

In certain embodiments, when the antisense strand of the RNA inhibitor 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 2'-F modified, and the rest are 2'-OMe modified, and there is a phosphorothioate connection between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides.

In certain embodiments, the RNA inhibitor, wherein

a) When the length of the sense strand is 21 nt, counting from the 5' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides of the sense strand, there is a phosphorothioate connection between the 2nd and 3rd nucleotides, the 9th, 10th, 11th, and 18th nucleotides of the sense strand are 2'-F modified, and the rest are 2'-OMe modified; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides of the sense strand, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides of the sense strand;

b) When the antisense chain is 26 nt in length, counting from the 5' end, the nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense chain are modified with 2'-F, and the rest are modified with 2'-OMe, and there is a phosphorothioate connection between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides.

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

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

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

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

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

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.

The modified sequences are shown in Table 2.

Table 2 Modified duplexes

Among them, 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 adjacent nucleotides to the left and right of * are connected by thiophosphate groups. (D02)*, (InvAB)*, (M06)*, *(D02), *(InvAB), *(M06) represent that D02, invAB, M06 are connected to nucleotides through thiophosphates. invAB refers to an inverted abasic nucleotide bonded to the 5' end or 3' end of the nucleotide. (M06) refers to a structure bonded to the 5' end or 3' end of a monomer (such as a nucleotide) as described below:

(M06) It can be linked to the nucleotide via phosphate or via thiophosphate.

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

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

Ligand-coupled RNA inhibitors

Another aspect of the RNA inhibitors of the present application relates to the manner in which the interfering nucleic acid is coupled to a ligand to enhance the stability, activity, cellular distribution or cellular uptake of the RNAi agent.

In certain embodiments, the distribution, targeting or stability of the 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., cell or organ compartment, body tissue, organ or region)) compared to a species in which the ligand is not present.

The ligand can include naturally occurring substances, such as proteins (e.g., human serum albumin (HSA), low-density lipoprotein (LDL) or globulin); 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.

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

The sense strand and antisense strand comprised by the present application's RNA inhibitor can be conveniently and routinely prepared by the known techniques 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 techniques to prepare other oligonucleotides (such as thiophosphates and alkylated derivatives).

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.

In certain embodiments, the ligand conjugation method of the present invention is to couple the ligand structure to the 5' end and/or 3' end of the antisense chain, and/or the 5' end and/or 3' end of the sense chain.

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.

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

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

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

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

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

Pharmaceutical composition

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

In one embodiment, provided herein is a pharmaceutical composition comprising an RNA inhibitor as described herein and a pharmaceutically acceptable pharmaceutical excipient.

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. An example scheme is a composition formulated for systemic administration with parenteral delivery, e.g., subcutaneous (SC), intramuscular (IM) or intravenous (IV) delivery. The pharmaceutical compositions of the present application can be administered at a dose sufficient to inhibit IGF-1R gene expression.

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

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

In certain embodiments, the delivery vehicle comprises a liposome.

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

use

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 the preparation of a drug for preventing or treating a disease or pathology or reducing the risk of a disease or pathology.

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

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

On the other hand, 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.

In certain embodiments, the RNA inhibitor that inhibits IGF-1R gene expression, its pharmaceutically acceptable salt 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.

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

In certain embodiments, the local or focal 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.

In certain embodiments, the eye tissue is an 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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

In certain embodiments, the ocular tissue is retinal ganglion, endothelial cell, periocular muscle or periocular fat.

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

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

As used herein, the term "inhibit" is used interchangeably with "reduce," "reduced," "silence," "downregulate," "suppress," and other similar terms, and includes any level of inhibition. Expression of the IGF-1R gene can be evaluated based on the level or change in level of any variable associated with IGF-1R gene expression, for example, IGF-1R mRNA level or IGF-1R protein level. This level can be analyzed in a single cell or in a population of cells (including, for example, a sample from a subject). Inhibition can be evaluated 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 baseline level before administration or a level measured from a similar subject, cell, or sample that has not been treated or has been treated with a control (such as, for example, a buffer-only control or a no active agent control).

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 the RNA inhibitor of the present application, or by administering the RNA inhibitor of the present application to a subject in which the cells are present) so that the IGF-1R gene expression is inhibited, compared to a second cell or cell population that is substantially the same as the first cell or cell population but has not been treated in this way (control cells that have not been treated with the RNA inhibitor or have not been treated with the RNA inhibitor targeting the gene of interest). In a preferred embodiment, inhibition is evaluated in a cell line with high gene expression of IGF-1R using an appropriate concentration of siRNA by the method provided in the Examples, and the mRNA level in the intervened cells is expressed as a percentage of the mRNA level in the non-intervention control cells.

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.

Inhibition of IGF-1R protein expression can be represented by a decrease in the level of IGF-1R protein expressed by a cell or cell population or a sample of a subject (e.g., the level of protein in a blood sample derived from a subject). As described above, for evaluation 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 a change in protein levels in a sample of a subject (e.g., blood or serum derived therefrom).

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

The level of IGF-1R expressed by a cell or cell population can be determined by any method known in the art for evaluating mRNA gene expression. For example, qRT-PCR evaluates the reduction of gene expression. The reduction of protein production can be evaluated by any method known in the art, for example, ELISA. In certain embodiments, a puncture liver biopsy sample is used as a tissue material for monitoring the reduction of IGF-1R gene or protein gene expression. In other embodiments, a blood sample is used as a subject sample for monitoring the reduction of IGF-1R protein expression.

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.

This application provides the following implementation modes:

1. An RNA inhibitor for inhibiting insulin-like growth factor 1 receptor (IGF-1R) gene expression, comprising an antisense chain, wherein the antisense chain 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, and more preferably 17-23 nucleotide pairs in length.

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

Nucleotide No. 1118, Nucleotide No. 1120, Nucleotide No. 1154, Nucleotide No. 1367, Nucleotide No. 1407, Nucleotide No. 1415, Nucleotide No. 1532, Nucleotide No. 1625, Nucleotide No. 1627, Nucleotide No. 1628, Nucleotide No. 1631, Nucleotide No. 3356, Nucleotide No. 3357, Nucleotide No. 3359, Nucleotide No. 3792, Nucleotide No. 4198, Nucleotide No. 4200, Nucleotide No. 4208, Nucleotide No. 4685, Nucleotide No. 5246, Nucleotide No. 5392, Nucleotide No. 5393, Nucleotide No. 6329, Nucleotide No. 6332, Nucleotide No. 6333, Nucleotide No. 6334 nucleotide No. 4, 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. 53 nucleotide at position 80, nucleotide at position 5381, nucleotide at position 5663, nucleotide at position 5700, nucleotide at position 5711, nucleotide at position 5712, nucleotide at position 5721, nucleotide at position 5896, nucleotide at position 5898, nucleotide at position 6203, nucleotide at position 6236, nucleotide at position 6237, nucleotide at position 6240, nucleotide at position 6245, nucleotide at position 6288, nucleotide at position 6299, nucleotide at position 6305, nucleotide at position 6309, nucleotide at position 6310, nucleotide at position 6343, nucleotide at position 6363, nucleotide at position 6365, nucleotide at position 6366, nucleotide at position 6373, nucleotide at position 6379, nucleotide at position 6381 Acid, 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, nucleotide No. 8975, nucleotide No. 8977, nucleotide No. 9163, nucleotide No. 9 nucleotide No. 175, 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. 10266, nucleotide No. 10490, nucleotide No. 10526 , nucleotide No. 10528, nucleotide No. 10534, nucleotide No. 10535, nucleotide No. 10537, nucleotide No. 10540, nucleotide No. 10541, nucleotide No. 10679, nucleotide No. 10694, nucleotide No. 10717, nucleotide No. 10728, nucleotide No. 10729, nucleotide No. 10730, nucleotide No. 10754, nucleotide No. 10763, nucleotide No. 10766, nucleotide No. 10942, nucleotide No. 11034, nucleotide No. 11078, nucleotide No. 11126, nucleotide No. 11156, nucleotide No. 11159, nucleotide No. 11160, nucleotide No. 11292, Nucleotide at position 11340, nucleotide at position 11344, nucleotide at position 11383, nucleotide at position 11395, nucleotide at position 11789, nucleotide at position 11864, nucleotide at position 11865, nucleotide at position 11939, nucleotide at position 12092, nucleotide at position 12116, nucleotide at position 12130, nucleotide at position 12154, nucleotide at position 12156, nucleotide at position 12177, nucleotide at position 12195, nucleotide at position 1164, nucleotide at position 1163, nucleotide at position 1162, nucleotide at position 1161, nucleotide at position 1160, nucleotide at position 1165, nucleotide at position 1166, nucleotide at position 1167, and nucleotide at position 1168.

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

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.

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

6. An RNA inhibitor according to any one of embodiments 1-5, which comprises a sense strand capable of forming a complementary double strand 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, and preferably the duplex complementary region is 15-30 nucleotide pairs in length, more preferably 17-23 nucleotide pairs in length.

7. The RNA inhibitor according to embodiment 6, wherein the sense strand and the antisense strand are present on two different nucleic acid strands, and 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 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.

9. An 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.

10. An 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.

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

12. An RNA inhibitor according to 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.

13. An RNA according to any one of embodiments 1-12, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO: 199-400.

14. An RNA inhibitor according to any one of embodiments 6-13, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO: 1-198.

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-n49, ds-n50, ds-n5 1. 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, d s-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, d s-n106, ds-n107, ds-n108, ds-n109, ds-n110, ds-n111, ds-n112, ds-n113, ds-n114, ds-n115, ds-n116, ds-n117, d s-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-n 154, ds-n155, ds-n156, ds-n157, ds-n158, ds-n159, ds-n160, ds-n161, ds-n162, ds-n163, ds-n164, ds-n165, ds-n1 66. ds-n167, ds-n168, ds-n169, ds-n170, ds-n171, ds-n172, ds-n173, ds-n174, ds-n175, ds-n176, ds-n177, ds-n17 8. 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 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.

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- 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.

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.

19. An 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.

20. The RNA inhibitor according to embodiment 19 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.

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.

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

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

24. An 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, respectively, Z is G or modified G, and N comprises at least one nucleotide.

25. An RNA inhibitor according to any one of embodiments 19-24, wherein X2, Y, and Z are AAG, AUG, UUG, or UAG, respectively, from 3' to 5', or the above sequences that have been partially or fully modified.

26. An RNA inhibitor according to any one of embodiments 21-25, wherein X2, Y, and Z are AAG, AUG, UUG, or UAG, respectively, from 3' to 5', or the above sequences that have been partially or fully modified.

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

28. An 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 the above sequences that have been partially or fully modified.

29. An RNA inhibitor according to any one of embodiments 16-28, wherein the antisense chain comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:401-413.

30. The RNA inhibitor according to 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.

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.

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.

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 (VP), N6-methyladenosine (m6A) modification, 5-methylcytidine (m5C) modification, 3-methyluridine (m3U) modification, 5-methylureidine (m5U) modification, pseudouridine modification, 2-thioureidine (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, M06 modification, invAb modification or invAB modification.

34. The RNA inhibitor according to 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.

35. An RNA inhibitor according to embodiment 34, wherein the phosphate mimetics include 5'-(E)-vinyl phosphonate, 5'-methyl phosphonate, (S)-5'-C-methyl analogs and 5'-thiophosphate (5'-PS).

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

37. The RNA inhibitor according to any one of embodiments 31-36, wherein the 3' or 5'-end of the antisense strand is modified with invAB, or the 3' or 5'-end of the sense strand is modified with invAB.

38. The RNA inhibitor according to any one of embodiments 31-37, 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.

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

a) the positive 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;

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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides.

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

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

42. The RNA inhibitor according to any one of embodiments 39-41, having the following modified combinations

a) The forward strand is 21 nt long and includes the following modifications: Counting from the 5' end, nucleotides at positions 9, 10, and 11 have 2'-F modifications;

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 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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides.

43. An RNA inhibitor according to any one of embodiments 28-42, wherein the antisense chain comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:624-812.

44. An RNA inhibitor according to any one of embodiments 28-42, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:414-602.

45. The RNA inhibitor according to 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, -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, d s-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-m6 1. 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-m 74. 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-m1 22. 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, d s-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-m 167, ds-m168, ds-m169, ds-m170, ds-m171, ds-m172, ds-m173, ds-m174, ds-m175, ds-m176, ds-m177, ds-m17 8. 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 embodiments 28-39, wherein

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, there is a phosphorothioate linkage between the 9th, 10th, 11th, and 18th nucleotides of the sense strand

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 there is a phosphorothioate linkage between the 2nd and 3rd nucleotides;

b) the antisense strand comprises a fourth sequence with a length of 19-26 nt, wherein the fourth sequence includes the following modifications: counting from the 5' end, the nucleotides at positions 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 connection between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides.

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

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

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

a) The length of the sense strand is 21 nt. Counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides. The 9th, 10th, 11th, and 18th nucleotides 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 the 1st and 2nd nucleotides, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides;

b) The antisense chain is 26 nt in length. Counting from the 5' end, the nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense chain are modified with 2'-F, and the rest are modified with 2'-OMe. 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.

50. An RNA inhibitor according to any one of embodiments 31-49, wherein the antisense chain comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:813-827.

51. An RNA inhibitor according to any one of embodiments 31-50, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO: 603-617.

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.

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 strand and/or antisense strand, and the delivery system enables the RNA inhibitor to reach the target RNA in the target tissue to produce a gene silencing effect.

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.

55. According to the RNA inhibitor described in embodiment 54, the eye tissue is 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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

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

57. An 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.

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.

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

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

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

62. An RNA inhibitor according to 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.

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

64. An 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 chain or antisense nucleic acid fragment.

65. An 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.

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 chain or the antisense nucleic acid fragment, and the ligand is conjugated to the 3’ end of the sense chain or the sense nucleic acid fragment, and the two ligands are the same or different.

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 chain or the antisense nucleic acid fragment, and the ligand is conjugated to the 5’ end of the sense chain or the sense nucleic acid fragment, and the two ligands are the same or different.

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.

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 chain or the antisense nucleic acid fragment, and the ligand is conjugated to the 3’ end of the sense chain or the sense nucleic acid fragment, and the two ligands are the same or different.

70. According to any one of embodiments 53-69, the number of delivery systems is 1, 2, 3, 4, 5 or 6.

71. An RNA inhibitor according to any one of embodiments 53-70, wherein the antisense chain comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:822-827.

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

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

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.

75. A composition according to embodiment 74, characterized in that the pharmaceutically acceptable carrier includes or is selected from an aqueous carrier, a liposome, a high molecular polymer or a polypeptide.

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.

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.

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

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.

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.

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

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

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.

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.

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, intra-articular injection, intrathecal injection, subcutaneous injection, intravenous injection, oral administration, rectal administration, or intraperitoneal administration.

86. The method according to any one of embodiments 83-85 comprises administering to a local or focal area tissue of a subject in need thereof.

87. The method according to embodiment 86, wherein the local or focal 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.

88. The method according to embodiment 87, wherein the eye tissue is an 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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

89. A method according to embodiment 88, wherein the eye tissue is retinal ganglion, endothelial cells, peripheral ocular muscle or peripheral ocular fat.

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.

91. A method according to any one of embodiments 83-90, wherein the IGF-1R related diseases or pathologies include thyroid eye disease, osteoarthritis, and 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 described in any one of embodiments 1-73 or the pharmaceutical composition described in any one of embodiments 74-75.

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

94. A method according to embodiment 93, wherein the disease or pathology includes thyroid eye disease, osteoarthritis, and neuropathic pain.

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.

96. A method according to embodiment 95, 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 injection, intravenous injection, oral administration, rectal administration or intraperitoneal administration.

97. The method according to any one of embodiments 95-96 comprises administering to a local or focal area tissue of a subject in need thereof.

98. A method according to embodiment 97, wherein the local or focal 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.

99. The method according to embodiment 98, wherein the eye tissue is an optic nerve, a trabecular meshwork, a proximal canal tissue, a ganglion, an episcleral vein, a 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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue.

100. A method according to embodiment 99, wherein the eye tissue is retinal ganglion, endothelial cells, peripheral ocular muscle or peripheral ocular fat.

Example

Example 1 Design and synthesis of siRNA molecules

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).

Oligoribonucleotides were synthesized using the phosphoramidite solid phase synthesis technique. The synthesis was performed on universal controlled pore glass (CPG). All 2'-modified RNA phosphoramidites and auxiliary reagents were commercially available reagents. 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. A 50 mM solution of 3-((dimethylamino-methylene)amino)-3H-1,2,4-dithiazole-3-thione in anhydrous acetonitrile/pyridine (v/v = 1/1) was used to generate phosphorothioate bonds. The reaction time was 1.8 min. All sequences were synthesized after the final removal of the DMT group.

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

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

Annealing of single-stranded oligoribonucleotides to produce siRNA: The single-stranded oligoribonucleotides to be annealed are prepared into 200μM with sterile RNase-free water. The annealing reaction system is set up as follows: a total volume of 100μL of the mixture, 10nmol, is placed in a 95℃ water bath for 10 minutes (≥100nmol requires high temperature for 20 minutes) → quickly placed in a 60℃ water bath, cooled naturally → the solution after annealing is not stored at high temperature. Complementary chains are formed by combining equimolar single-stranded oligoribonucleotide solutions. The unmodified siRNA sequence is shown in Table 1.

The siRNA duplex was subjected to sequence modification optimization, wherein lowercase letters "g", "c", "a" and "u" represent 2'-methoxy-modified nucleotides; uppercase letters "Gf", "Cf", "Af" and "Uf" represent 2'-fluoro-modified nucleotides; * represents that the two monomers (such as two nucleotides) adjacent to the left and right of * are connected by a phosphorothioate group.

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

In the present invention, (M06) It is M06 monomer In certain embodiments, (M06) is represented by Bonded to a nucleotide.

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

The modified siRNA sequences are shown in Table 2.

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

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

1) SY5Y cell transfection

SY5Y cells were cultured in DMEM medium containing 10% fetal bovine serum, 1% glutamine, 1% NEAA, and 1% penicillin-streptomycin in a 5% CO2, 37°C constant temperature incubator, and transfected when the cells were in the logarithmic growth phase and in good condition. SY5Y cells were inoculated into 96-well cell plates, plated, and siRNA was transfected into the cells using RNAiMAX according to the instructions. The final concentrations of siRNA in this test were 30nM and 0.1nM. A compound-free control group containing RNAiMAX was also set up.

2) RNA extraction and reverse transcription

24 hours after transfection, remove the culture medium and collect the cells for RNA extraction. Use according to the kit instructions. Total RNA was extracted using 96 Kit (QIAGEN-74182), and cDNA was synthesized using FastKing RT Kit (With gDNase) (Tiangen-KR116-02) according to the instruction manual.

3) qPCR detection of target gene mRNA expression level

The target cDNA will be detected by SYBR Green qPCR, and GAPDH cDNA will be detected in parallel as an internal control. 8μL of the prepared PCR reaction solution and 2μL of sample cDNA are added to the 384 wells. The qPCR reaction program is: heating at 50℃ for 2min, heating at 95℃ for 10min and then entering the cycle mode, heating at 95℃ for 15sec, followed by 1min at 60℃, for a total of 40 cycles.

4) Result analysis

The expression level of target gene mRNA in each sample was calculated by ΔΔCt relative quantification method. The relative amount of target gene was expressed as 2-ΔΔCT.

The calculation formula is as follows:

ΔCT = average Ct value of target gene - average Ct value of reference gene

ΔΔCT＝ΔCT(drug-added group)-ΔCT(RNAiMAX control group)

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

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

The results are shown in Table 3.

Table 3 Inhibition results of duplex siRNA on SY5Y (percent inhibition rate)

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

Example 3 In vitro effect detection of chemically modified duplex siRNA on Hela cells

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

1) Hela cell transfection

Hela cells were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin in a 5% CO2, 37°C constant temperature incubator, and transfected when the cells were in the logarithmic growth phase and in good condition. Hela cells were inoculated into 96-well cell plates, plated, and double-stranded siRNA was transfected into Hela cells using lipofectamine RNAiMAX and Opti-MEM according to the instructions. The final concentrations of siRNA in this test were 30nM, 5nM, and 0.5nM.

2) RNA extraction, reverse transcription and qPCR

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

3) qPCR detection of target gene mRNA expression level

The experimental steps are the same as in Example 2.

4) Result analysis

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

The results are shown in Table 4.

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

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

Example 4 In vitro effect detection of chemically modified duplex siRNA on ARPE cells

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

1) ARPE cell transfection

ARPE-19 cells were cultured in DMEM medium containing 10% fetal bovine serum in a 5% CO2, 37°C constant temperature incubator, and transfected when the cells were in the logarithmic growth phase and in good condition. ARPE-19 cells were inoculated into 96-well cell plates, plated, and double-stranded siRNA was transfected into Hela cells using lipofectamine RNAiMAX and Opti-MEM according to the instructions. The final concentrations of siRNA in this test were 30nM, 0.5nM, and 0.05nM.

2) RNA extraction, reverse transcription and qPCR

The experimental steps are the same as in Example 2.

3) qPCR detection of target gene mRNA expression level

The experimental steps are the same as in Example 2.

4) Result analysis

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

The results are shown in Table 5.

Table 5 Inhibition results of duplex siRNA on ARPE (percent inhibition rate)

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

Example 5 IC ₅₀ Detection of Chemically Modified Duplex siRNA in Primary Human Mature Adipocytes

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

5) Transfection of primary human mature adipocytes

Primary human mature adipocytes were cultured in a 5% CO ₂ , 37°C constant temperature incubator in a special adipocyte culture medium, and transfected when the cells were in the logarithmic growth phase and in good condition. Mature adipocytes were inoculated into 24-well cell plates, plated, and double-stranded siRNA was transfected into mature adipocytes using lipofectamine RNAiMAX and Opti-MEM according to the instructions.

6) RNA extraction, reverse transcription and qPCR

The experimental steps are the same as in Example 2.

7) qPCR detection of target gene mRNA expression level

The experimental steps are the same as in Example 2.

8) Result analysis

The results are shown in Table 6.

Table 6 Inhibition results of duplex siRNA on primary mature human adipocytes

The results showed that the above double-stranded siRNA can significantly inhibit the level of IGF-1R mRNA in human primary mature adipocytes.

Example 6 PD study of intraorbital administration in cynomolgus monkeys

Group design

Six female cynomolgus monkeys were divided into two groups. Before administration, a biopsy was performed on the left eye to collect a small amount of eye muscle and periorbital fat. The collected samples were placed separately in homogenized tubes containing 0.5 mL of RNAlater (2 tubes of eye muscle and 1 tube of fat), and placed in a refrigerator at 2°C–8°C for about 22-26 hours, after which the RNAlater was completely removed. The homogenized tubes containing the tissues were temporarily stored on dry ice for quick freezing 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, 10 mg/kg) of the test article Z4 were administered to the right eye.

Test Procedure

1 Clinical observation

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.

2 weight

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

3. Ophthalmological examination

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

4. Intraocular pressure

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.

5 Fundus Photography (FP)

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

6Flash visual evoked potential (fVEP)

All animals will be anesthetized with an intramuscular injection of ketamine (10-30 mg/kg) and xylazine (0.5 to 1.0 mg/kg). The animal's pupils will be dilated with an appropriate mydriatic. Flash VEP is performed as follows:

Select the FVEP protocol and enter the animal information;

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

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

After judging the inspection results, save the inspection report.

7 Cytokine Analysis

Blood samples will be collected from all available animals for analysis of cytokines in serum. The collection time points are once before administration and once 24 hours after administration. The cytokine levels of IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, TNF-α and IFN-γ in serum will be analyzed using a validated flow cytometry method.

End point anatomy

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

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.

IGF1R mRNA QPCR analysis

The mRNA expression of IGF1R was detected in cynomolgus monkey tissues by RT-qPCR using the dye method. The CT values of the target gene (IGF1R) and the reference gene (CypA) were detected to calculate the expression changes of the target gene. Tissue types included: periocular muscle and periocular fat.

1Key primer reagents

2 Key Equipment

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

CyBio-SELMA liquid handling workstation, UV spectrophotometer, TECAN liquid handling workstation

3RNA extraction

1000μL TRNzol Universal RNA Reagent was added to each tissue sample, followed by 3 grinding beads (3mm). The homogenizer program was set to: 65Hz, 45s/cycle, 4 cycles, 30s between cycles (or 5.65m/s, 1min/cycle, 3 cycles, 30s between cycles) to obtain tissue lysate. Each tissue lysate was centrifuged (centrifugation conditions: room temperature, 12000×g, 5min), and all supernatants were transferred to new Ep tubes after centrifugation. Incubate at room temperature for 5 minutes to completely dissociate the nucleoprotein complex. 200μL chloroform was added to each tissue lysate, vortexed to mix, centrifuged instantly, and incubated at room temperature for 3min. Centrifugation was performed (centrifugation conditions: 4℃, 12000×g, 5min). After centrifugation, the tissue lysate was divided into three layers of water phase: upper, middle, and lower. 450μL of the upper water phase was transferred to a new Ep tube for each portion. Add 45μL 3M Sodium Acetate Sol pH 5.2 to each portion, vortex to mix, and incubate at room temperature for 1 min. After incubation, add 500μL isopropanol (pre-cooled at low temperature), vortex to mix, and incubate at -30℃～-10℃ for 30min. After the incubation, centrifuge (centrifugation conditions: 4℃, 12000×g, 15min). After the centrifugation, RNA precipitation can be seen at the bottom of the tube, and the supernatant is discarded. After discarding the supernatant, add 1000μL 75% ethanol solution (pre-cooled at low temperature), vortex to mix, and centrifuge (centrifugation conditions: 4℃, 12000×g, 10min). After centrifugation, discard the supernatant, air-dry at room temperature for 10min, then add 40μL DEPC-treated water (pre-heated at 55℃), vortex to mix, and obtain RNA. Use a UV spectrophotometer to detect A260/A280 to obtain purity and concentration and record them. RNA can be directly used for RT-qPCR or stored at <-60°C. The expiration date of RNA is 1 year.

4QPCR analysis

The RNA was run through the genomic DNA removal program to obtain the "RNA template (after removing DNA), and then run through the "PCR thermal cycler program to obtain the cDNA product as the "qPCR template". All cDNA samples were tested by qPCR using a 384-well plate, and all were analyzed in triplicate. Relative expression data will be collected using QuantStudio7 Flex PCR, and the collected data include: Ct average, ΔCt, ΔΔCt. All IGF1R mRNA KD% was calculated after normalization with the biopsy sample.

Experimental results:

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

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

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

Example 7: Acute exophthalmos model in mice

Objective: To evaluate the efficacy of mouse IGF1R siRNA Z8, Z9 delivered intraorbitally in the BALB/c mouse thyroid eye disease model.

1 Group dosing regimen

On Day 5, after the animals were anesthetized with Zotal (25-50 mg/kg, i.p.) and Xylazine Hydrochloride (5 mg/kg, i.p.), a single injection of mouse IGF1R siRNA was given into both eye orbits, 100 μL/eye.

2 Modeling

On Day 0 and Day 6, the animals were anesthetized with Zotal (25-50 mg/kg, i.p.) and Xylazine Hydrochloride (5 mg/kg, i.p.), and mouse IGF-1 protein was injected periocularly in both eyes for modeling, 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.

3. Testing indicators

3.1 Clinical observation

Observe the animal's condition, mental state, behavioral activities, eating habits, etc. once a day beside the cage.

3.2 GO (Graves' Ophthalmopathy) score

GO scores were performed on Day 0 (before modeling), Day 3, Day 6 (before modeling), and Day 9, and the symptoms of bulging eyes were continuously observed.

3.3 Eye examination

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 recorded by photograph.

3.4 Collection of materials

On Day 9, 4 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 preservation solution.

3.5 qPCR detection

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

3.6 Pathology

On Day 9, eyeballs (with periocular tissues) of 4 mice were collected from each group for fixation and paraffin embedding, and the proliferation and cell infiltration of periocular tissues were quantified (H&E staining, UCPI and CD3 staining).

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

Experimental Example 7: Rat surgically induced osteoarthritis model

At 10 weeks after birth, 24 rats underwent surgical transection of the anterior cruciate ligament, medial collateral ligament, and medial meniscocondylar ligament (ACLT+pMMx model). 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. Sections from the anterior part (5 μm thickness, 100 μm interval between each section to ensure reproducibility) were taken at different levels and stained with safranin fast green. At least 12 sections were taken from each rat and imaged using a light microscope.

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

Histological evaluation was performed by two blinded observers. Images were scored according to the OARSI cartilage histological scoring system by assessing the extent of cartilage damage (depth of cartilage damage) and stage (extent of joint involvement). Briefly, femurs and tibiae were evaluated separately, and each site was scored according to the grade of cartilage damage (0–6 points, 0 is intact surface and 6 is severe deformation) and the stage of cartilage damage (0–4 points, 0 is normal joint and 4 is more than 50% joint damage). The total score was the product of grade and stage (0 represents normal joint; 24 represents severe osteoarthritis). Twelve sections were scored for each rat, and no rats were excluded. After scoring, the study was unblinded, and the four lowest-scoring sections (representing the least cartilage damage) of each rat were excluded for 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.

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 involved in the present invention can effectively treat osteoarthritis in humans.

Example 8: Spinal Nerve Ligation (SNL) Model

Animals were first anesthetized with 5% isoflurane and maintained at 2% isoflurane concentration. 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 group, the L4 spinal nerve was exposed but not ligated. Under isoflurane anesthesia, mouse IGF1R siRNA Z8, Z9 or control solution was injected by lumbar puncture and subsequent behavioral tests were performed.

Behavioral testing

50% Paw Retraction Mechanical Threshold (PWT):

Acclimation period: The rats were allowed to acclimate to the test area for at least three days before measurements were taken. The lateral plantar aspect was stimulated vertically using von Frey fibers of different 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 taken one day before surgery and on the 4th, 7th, 10th, and 14th days after surgery. The 4.0 g fiber was selected first, and then the 50% PWT threshold of the rats was calculated according to the “up-down method”. The proportion of paw withdrawal in the rats to increasing mechanical stimulation was measured. The three groups of rats were evaluated sequentially, and five measurements were taken on each side (5 min intervals between each). Finally, the five paw withdrawal response rates, i.e., the proportion of paw withdrawal produced by the rats to the 6.0 g mechanical stimulation, were calculated.

Cold hyperalgesia threshold:

To assess the sensitivity of the foot to cold, the rat's foot response to acetone was measured. Precooled acetone (100 μL) was applied to the surface of the sole of the foot while the rat was in a quiet state. The time of positive reactions (lifting the foot, shaking, licking, etc.) was recorded for 30 seconds. Each foot was tested three times (5 minutes apart each time), and the average of the three measurements was recorded as the reaction time of that side.

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

Claims (100)

An RNA inhibitor for inhibiting the expression of insulin-like growth factor 1 receptor (IGF-1R) gene, comprising an antisense chain, wherein the antisense chain 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, and more preferably 17-23 nucleotide pairs in length. The RNA inhibitor according to claim 1, wherein the antisense strand and the mRNA encoding IGF-1R (SEQ ID NO: 828) form a complementary region starting from any of the following positions counting from the 5' end: Nucleotide No. 1118, Nucleotide No. 1120, Nucleotide No. 1154, Nucleotide No. 1367, Nucleotide No. 1407, Nucleotide No. 1415, Nucleotide No. 1532, Nucleotide No. 1625, Nucleotide No. 1627, Nucleotide No. 1628, Nucleotide No. 1631, Nucleotide No. 3356, Nucleotide No. 3357, Nucleotide No. 3359, Nucleotide No. 3792, Nucleotide No. 4198, Nucleotide No. 4200, Nucleotide No. 4208, Nucleotide No. 4685, Nucleotide No. 5246, Nucleotide No. 5392, Nucleotide No. 5393, Nucleotide No. 6329, Nucleotide No. 6332, Nucleotide No. 6333, Nucleotide No. 6334 nucleotide No. 4, 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. 53 nucleotide at position 80, nucleotide at position 5381, nucleotide at position 5663, nucleotide at position 5700, nucleotide at position 5711, nucleotide at position 5712, nucleotide at position 5721, nucleotide at position 5896, nucleotide at position 5898, nucleotide at position 6203, nucleotide at position 6236, nucleotide at position 6237, nucleotide at position 6240, nucleotide at position 6245, nucleotide at position 6288, nucleotide at position 6299, nucleotide at position 6305, nucleotide at position 6309, nucleotide at position 6310, nucleotide at position 6343, nucleotide at position 6363, nucleotide at position 6365, nucleotide at position 6366, nucleotide at position 6373, nucleotide at position 6379, nucleotide at position 6381 Acid, 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, nucleotide No. 8975, nucleotide No. 8977, nucleotide No. 9163, nucleotide No. 9 nucleotide No. 175, 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. 10266, nucleotide No. 10490, nucleotide No. 10526 , nucleotide No. 10528, nucleotide No. 10534, nucleotide No. 10535, nucleotide No. 10537, nucleotide No. 10540, nucleotide No. 10541, nucleotide No. 10679, nucleotide No. 10694, nucleotide No. 10717, nucleotide No. 10728, nucleotide No. 10729, nucleotide No. 10730, nucleotide No. 10754, nucleotide No. 10763, nucleotide No. 10766, nucleotide No. 10942, nucleotide No. 11034, nucleotide No. 11078, nucleotide No. 11126, nucleotide No. 11156, nucleotide No. 11159, nucleotide No. 11160, nucleotide No. 11292, Nucleotide at position 11340, nucleotide at position 11344, nucleotide at position 11383, nucleotide at position 11395, nucleotide at position 11789, nucleotide at position 11864, nucleotide at position 11865, nucleotide at position 11939, nucleotide at position 12092, nucleotide at position 12116, nucleotide at position 12130, nucleotide at position 12154, nucleotide at position 12156, nucleotide at position 12177, nucleotide at position 12195, nucleotide at position 1164, nucleotide at position 1163, nucleotide at position 1162, nucleotide at position 1161, nucleotide at position 1160, nucleotide at position 1165, nucleotide at position 1166, nucleotide at position 1167, and nucleotide at position 1168. An RNA inhibitor according to any one of claims 1-2, wherein the antisense strand and the mRNA encoding IGF-1R (SEQ ID NO:828) form a complementary region between the 1160th nucleotide and the 1188th nucleotide, between the 6235th nucleotide and the 6465th nucleotide, or between the 6329th nucleotide and the 6455th nucleotide, counting from the 5' end. The RNA inhibitor according to any one of claims 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. The RNA inhibitor according to any one of claims 1 to 4, wherein the RNA inhibitor is an antisense oligonucleotide (ASO), shRNA, miRNA or siRNA. The RNA inhibitor according to any one of claims 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, the duplex complementary region having 0, 1, 2, 3, 4 or 5 mismatches, and preferably the duplex complementary region has a length of 15-30 nucleotide pairs, more preferably 17-23 nucleotide pairs. 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. 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, and preferably the RNA inhibitor is shRNA. The RNA inhibitor according to any one of claims 6 to 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. 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. 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. 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. The RNA according to any one of claims 1-12, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO: 199-400. An RNA inhibitor according to any one of claims 6-13, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO: 1-198. The RNA inhibitor according to any one of claims 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, 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, d s-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-n6 5. 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-n 79. 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-n 130, ds-n131, ds-n132, ds-n133, ds-n134, ds-n135, ds-n136, ds-n137, ds-n138, ds-n139, ds-n140, ds-n141, ds-n 142, ds-n143, ds-n144, ds-n145, ds-n146, ds-n147, ds-n148, ds-n149, ds-n150, ds-n151, ds-n152, ds-n153, ds-n1 54. ds-n155, ds-n156, ds-n157, ds-n158, ds-n159, ds-n160, ds-n161, ds-n162, ds-n163, ds-n164, ds-n165, ds-n1 66. ds-n167, ds-n168, ds-n169, ds-n170, ds-n171, ds-n172, ds-n173, ds-n174, ds-n175, ds-n176, ds-n177, ds-n17 8. 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. 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-n9 8. 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. The RNA inhibitor according to 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-n 83. 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. The RNA inhibitor according to claim 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. The RNA inhibitor according to any one of claims 1 to 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. The RNA inhibitor according to claim 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. The RNA inhibitor according to any one of claims 16 to 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. The RNA inhibitor according to claim 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. 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. The RNA inhibitor according to any one of claims 19 to 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, respectively, the Z is G or modified G, and the N comprises at least one nucleotide. The RNA inhibitor according to any one of claims 19 to 24, wherein X2, Y, and Z are AAG, AUG, UUG, or UAG in sequence from 3' to 5', or partially or fully modified versions of the above sequences. The RNA inhibitor according to any one of claims 21 to 25, wherein X2, Y, and Z are AAG, AUG, UUG, or UAG in sequence from 3' to 5', or partially or fully modified versions of the above sequences. The RNA inhibitor according to any one of claims 21 to 26, wherein N is C or modified C. The RNA inhibitor according to any one of claims 21 to 27, wherein 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. An RNA inhibitor according to any one of claims 16-28, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:401-413. The RNA inhibitor according to any one of claims 1 to 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. The RNA inhibitor according to any one of claims 1 to 30, wherein at least one nucleotide in the RNA inhibitor is a modified nucleotide. 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. The RNA inhibitor according to any one of claims 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, such as 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. The RNA inhibitor according to any one of claims 31 to 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. The RNA inhibitor of claim 34, wherein the phosphate mimic comprises 5'-(E)-vinylphosphonate, 5'-methylphosphonate, (S)-5'-C-methyl analogs and 5'-phosphorothioate (5'-PS). The RNA inhibitor according to any one of claims 31 to 35, wherein the 5'-end of the antisense strand comprises (M06) modification: The RNA inhibitor according to any one of claims 31 to 36, wherein the 3' or 5' end of the antisense strand is modified with invAB, or the 3' or 5' end of the sense strand is modified with invAB. The RNA inhibitor according to any one of claims 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. The RNA inhibitor according to any one of claims 31 to 38, having the following modification combination a) the positive 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; 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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides. The RNA inhibitor according to claim 39, wherein the length of the first sequence is preferably 16nt, 17nt, 18nt, 19nt, 20nt, 21nt, preferably 21nt. The RNA inhibitor according to claim 39, wherein the length of the second sequence is preferably 16nt, 17nt, 18nt, 19nt, 20nt, 21nt, preferably 21nt. The RNA inhibitor according to any one of claims 39 to 41, having the following modification combination a) The forward strand is 21 nt long and includes the following modifications: Counting from the 5' end, nucleotides at positions 9, 10, and 11 have 2'-F modifications; 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 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 the 2nd, 3rd, 4th, 12th, 14th, and 16th nucleotides. An RNA inhibitor according to any one of claims 28-42, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:624-812. An RNA inhibitor according to any one of claims 28-42, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:414-602. The RNA inhibitor according to any one of claims 28 to 44, which 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-m1 4. 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, d s-m113, ds-m114, ds-m115, ds-m116, ds-m117, ds-m118, ds-m119, ds-m120, ds-m121, ds-m122, ds-m123, d s-m124, ds-m125, ds-m126, ds-m127, ds-m128, ds-m129, ds-m130, ds-m131, ds-m132, ds-m133, ds-m134, d s-m135, ds-m136, ds-m137, ds-m138, ds-m139, ds-m140, ds-m141, ds-m142, ds-m143, ds-m144, ds-m145, d s-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. The RNA inhibitor according to any one of claims 28 to 39, wherein 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 connection between the 1st and 2nd nucleotides of the sense strand, there is a phosphorothioate connection between the 2nd and 3rd nucleotides, there is a 2'-F modification at the 9th, 10th, 11th, and 18th nucleotides of the sense strand, and the rest are 2'-OMe modified; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides; b) the antisense strand comprises a fourth sequence with a length of 19-26 nt, wherein the fourth sequence includes the following modifications: counting from the 5' end, the nucleotides at positions 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 connection between the 4th and 5th nucleotides; counting from the 3' end, there is a phosphorothioate connection between the 1st and 2nd nucleotides, and there is a phosphorothioate connection between the 2nd and 3rd nucleotides. 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, preferably 21 nt. The RNA inhibitor according to claim 47, wherein the length of the fourth sequence is preferably 19nt, 20nt, 21nt, 22nt, 23nt, 24nt, 25nt, 26nt, preferably 26nt. The RNA inhibitor according to any one of claims 46 to 48, wherein a) The length of the sense strand is 21 nt. Counting from the 5' end, there is a phosphorothioate linkage between the 1st and 2nd nucleotides of the sense strand, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides. The 9th, 10th, 11th, and 18th nucleotides 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 the 1st and 2nd nucleotides, and there is a phosphorothioate linkage between the 2nd and 3rd nucleotides; b) The antisense chain is 26 nt in length. Counting from the 5' end, the nucleotides at positions 1, 2, 5, 9, 17, and 19 of the antisense chain are modified with 2'-F, and the rest are modified with 2'-OMe. 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. An RNA inhibitor according to any one of claims 31-49, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:813-827. An RNA inhibitor according to any one of claims 31-50, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO: 603-617. The RNA inhibitor according to any one of claims 31 to 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-m 197, ds-m198, ds-m199, ds-m200, ds-m201, ds-m202, ds-m203 and ds-m204. 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 enables the RNA inhibitor to reach the target RNA of the target tissue to produce a gene silencing effect. The RNA inhibitor according to claim 53, wherein the target tissue is eye tissue, joint tissue, central nervous tissue, peripheral nervous tissue, tumor, liver tissue, kidney tissue, muscle tissue, and adipose tissue. According to the RNA inhibitor of claim 54, wherein the eye tissue is an optic nerve, a trabecular meshwork, a proximal canal tissue, a ganglion, an episcleral vein, a Schlemm's canal or an eye peripheral 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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue. The RNA inhibitor according to claim 55, wherein the eye tissue is retinal ganglion, endothelial cells, peripheral ocular muscle or peripheral ocular fat. The RNA inhibitor according to any one of claims 53 to 56, wherein the delivery system is independently conjugated to one or more internal positions of the double-stranded RNA. The RNA inhibitor of claim 57, wherein the internal position is on a nucleobase, a sugar ring, a methylphosphonate bond, a phosphorothioate diester bond, or a phosphodiester bond. The RNA inhibitor according to any one of claims 53 to 58, wherein the delivery system is a lipophilic structure, 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. The RNA inhibitor according to claim 59, wherein the lipophilic structure is selected from aliphatic, alicyclic and polyaliphatic compounds. The RNA inhibitor according to claim 60, wherein the lipophilic structure contains saturated or unsaturated C16 or C22. The RNA inhibitor according to any one of claims 59 to 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. The RNA inhibitor according to claims 53-62, wherein the delivery system is 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. 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. 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 the antisense nucleic acid fragment, and the ligand is conjugated to the 3' end of the sense strand or the sense nucleic acid fragment, and the two ligands are the same or different. 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 the antisense nucleic acid fragment, and the ligand is conjugated to the 5' end of the sense strand or the sense nucleic acid fragment, and the two ligands are the same or different. 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 the antisense nucleic acid fragment, and the ligand is conjugated to the 5' end of the sense strand or the sense nucleic acid fragment, and the two ligands are the same or different. 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 the antisense nucleic acid fragment, and the ligand is conjugated to the 3' end of the sense strand or the sense nucleic acid fragment, and the two ligands are the same or different. According to any one of claims 53-69, the number of delivery systems is 1, 2, 3, 4, 5 or 6. An RNA inhibitor according to any one of claims 53-70, wherein the antisense strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO:822-827. An RNA inhibitor according to any one of claims 53-71, wherein the positive strand comprises at least 15 consecutive nucleotides in a sequence as described in any one of SEQ ID NO: 618-623. The RNA inhibitor according to any one of claims 53 to 72, which is selected from: Z1, Z2, Z3, Z4, Z5, and Z6. A pharmaceutical composition comprising the RNA inhibitor according to any one of claims 1 to 73, and/or a physiologically acceptable excipient and/or carrier and/or diluent. The composition according to claim 74 is characterized in that the pharmaceutically acceptable carrier includes or is selected from aqueous carriers, liposomes, high molecular polymers or polypeptides. 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. The use according to claim 76, wherein the IGF-1R related disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels. The use according to any one of claims 76-77, wherein the IGF-1R related diseases or pathologies include thyroid eye disease, osteoarthritis, and neuropathic pain. The use according to any one of claims 76 to 78, wherein the RNA inhibitor and the pharmaceutical composition thereof are the RNA inhibitor according to any one of claims 1 to 73 or the pharmaceutical composition according to any one of claims 74 to 75. Use of the RNA inhibitor according to any one of claims 1 to 73 or the pharmaceutical composition according to any one of claims 74 to 75 in the preparation of a medicament for preventing or treating a disease or pathology or reducing the risk of a disease or symptom. The use according to claim 80, wherein the disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels. The use according to any one of claims 80-81, wherein the disease or pathology comprises thyroid eye disease, osteoarthritis, and neuropathic pain. A method for preventing or treating an IGF-1R-related disease or symptom comprises administering an effective amount of an RNA inhibitor and a pharmaceutical composition thereof to a subject in need thereof. The method according to claim 83, wherein the RNA inhibitor and the pharmaceutical composition thereof are the RNA inhibitor according to any one of claims 1-73 or the pharmaceutical composition according to any one of claims 74-75. The method of claims 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. The method according to any one of claims 83-85, comprising administering to a local or focal area tissue of a subject in need thereof. The method according to claim 86, wherein the local or focal 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. The method according to claim 87, wherein the eye tissue is an optic nerve, a trabecular meshwork, a proximal canal tissue, a ganglion, an episcleral vein, a 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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue. The method of claim 88, wherein the ocular tissue is retinal ganglion, endothelial cells, peripheral ocular muscle or peripheral ocular fat. The method according to any one of claims 83-89, wherein the IGF-1R related disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels. The method according to any one of claims 83-90, wherein the IGF-1R related diseases or pathologies include thyroid eye disease, osteoarthritis, and neuropathic pain. A method for inhibiting IGF-1R gene expression in a cell, tissue or subject, comprising administering an effective amount of the RNA inhibitor of any one of claims 1 to 73 or the pharmaceutical composition of any one of claims 74 to 75 to the cell, tissue or subject. The method of claim 92, wherein the disease or pathology comprises a disease or symptom associated with elevated IGF-1R levels. The method of claim 93, wherein the disease or pathology comprises thyroid eye disease, osteoarthritis, neuropathic pain. A method for preventing or treating a disease or symptom, the method comprising administering to a subject in need thereof 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. The method of claim 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. The method according to any one of claims 95-96, comprising administering to a local or focal area tissue of a subject in need thereof. The method according to claim 97, wherein the local or focal 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. The method according to claim 98, wherein the eye tissue is an optic nerve, a trabecular meshwork, a proximal canal tissue, a ganglion, an episcleral vein, a 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 nervous tissue and muscle nervous tissue; preferably, the adipose tissue comprises subcutaneous adipose tissue and visceral adipose tissue. The method of claim 99, wherein the ocular tissue is retinal ganglion, endothelial cells, peripheral ocular muscle or peripheral ocular fat.