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CN-121986165-A - Antisense oligonucleotide targeting SPTBN1 and application thereof

CN121986165ACN 121986165 ACN121986165 ACN 121986165ACN-121986165-A

Abstract

An antisense oligonucleotide aiming at Sptbnl targets and application thereof are provided. The antisense oligonucleotide can be used for knocking down Sptbnl expression, and can be used for treating hyperlipidemia and NASH and also can be used for treating neovascular eye diseases through in-vivo and in-vitro experiments.

Inventors

  • WANG HAISHENG
  • Lai fan
  • Dang Yunkun
  • WANG XIAOLEI
  • YU XIAOHUA
  • WANG YUHANG
  • CONG DEZI

Assignees

  • 思合基因(北京)生物科技有限公司
  • 思合(北京)生物医药有限公司

Dates

Publication Date
20260505
Application Date
20240628

Claims (20)

  1. An antisense oligonucleotide that inhibits SPTBN1 expression, wherein said antisense oligonucleotide targets SPTBN1 nucleic acid and is at least 96%, 97%, 98%, 99% complementary or fully (100%) complementary to SEQ ID No.1 or SEQ ID No. 2.
  2. The antisense oligonucleotide of claim 1, wherein said antisense oligonucleotide has 16-20 nucleotides.
  3. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide has a sequence set forth in SEQ ID NOs 4-16, 18-24, 26-34, 36-43, 48-54, 56-57, 60-68, 70-78.
  4. The antisense oligonucleotide of any one of claims 1-3, wherein at least one nucleoside in the oligonucleotide comprises a modified sugar or modified nucleobase.
  5. The antisense oligonucleotide of claim 4, wherein the modified sugar is a 2' -modified sugar.
  6. The antisense oligonucleotide of claim 5, wherein the 2 '-modified sugar is a 2' -O-methoxyethyl modification and/or a limiting ethyl modification.
  7. The antisense oligonucleotide of claim 4, wherein the modified nucleobase is a 5-methylcytosine.
  8. The antisense oligonucleotide of any one of claims 1-3, wherein at least one internucleoside linkage in the antisense oligonucleotide is a modified internucleoside linkage, preferably wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.
  9. The antisense oligonucleotide of claim 8, wherein each internucleoside linkage in the modified oligonucleotide is a phosphorothioate internucleoside linkage.
  10. The antisense oligonucleotide of any one of claims 1-3, wherein the antisense oligonucleotide comprises a gap comprised of linked deoxynucleosides and 5 'and 3' wing comprised of linked nucleosides, wherein the spacer is positioned between the 5 'and 3' wing, and wherein each nucleoside of each wing comprises a modified sugar.
  11. The antisense oligonucleotide of claim 10, wherein the antisense oligonucleotide comprises a gap consisting of 8-12 linked deoxynucleosides and 5 'and 3' wing segments consisting of 2-6 linked nucleosides, wherein the gap is between the 5 'and 3' wing segments and wherein each nucleoside of each wing segment comprises a modified sugar.
  12. The antisense oligonucleotide of claim 11, wherein the antisense oligonucleotide comprises a gap of 10 linked deoxynucleosides and 5' and 3' wing segments of 5 linked nucleosides, wherein the gap is between the 5' and 3' wing segments and wherein each nucleoside of each wing segment comprises a modified sugar, wherein each nucleoside of each wing segment is 2' -O-methoxyethyl sugar.
  13. The antisense oligonucleotide of claim 11, wherein the antisense oligonucleotide comprises a gap of 10 linked deoxynucleosides and 5 'and 3' wing segments of 3 linked nucleosides, wherein the gap is between the 5 'and 3' wing segments and wherein each nucleoside of each wing segment comprises a modified sugar, wherein each nucleoside of each wing segment is a limiting ethyl sugar.
  14. The antisense oligonucleotide of claim 12, wherein the antisense oligonucleotide consists of nucleosides having the nucleobase sequence of SEQ ID NOs 4-16, 18-24, 26-34, 36-43, and the single stranded modified oligonucleotide comprises a gap consisting of 10 linked deoxynucleosides and 5' and 3' wing consisting of 5 linked nucleosides, respectively, wherein the gap is between the 5' and 3' wing, and wherein each nucleoside of each wing is a 2' -O-methoxyethyl modified nucleoside, the internucleoside linkages in the entire modified oligonucleotide are phosphorothioate linkages, and all cytosines in the entire modified oligonucleotide are 5-methylcytosines.
  15. The antisense oligonucleotide of claim 13, wherein the antisense oligonucleotide consists of nucleosides having the nucleobase sequences of SEQ ID NOs 48-54, 56-57, 60-68, 70-78, and the single stranded modified oligonucleotide comprises a gap consisting of 10 linked deoxynucleosides and 5 'and 3' wing consisting of 3 linked nucleosides, respectively, wherein the gap is between the 5 'and 3' wing, and wherein each nucleoside of each wing is a limiting ethyl sugar, the internucleoside linkage in the entire modified oligonucleotide is a phosphorothioate linkage, and all cytosines in the entire modified oligonucleotide are 5-methylcytosines.
  16. The antisense oligonucleotide of any one of claims 1-15, wherein the antisense oligonucleotide comprises one or more ligands of an N-acetylgalactosamine (GalNAc) derivative.
  17. The antisense oligonucleotide of claim 16, wherein the ligand of the N-acetylgalactosamine (GalNAc) derivative is attached to the oligonucleotide by a linker.
  18. The antisense oligonucleotide of claim 17, wherein the linker is a monovalent, divalent, or trivalent branched linker.
  19. The antisense oligonucleotide of any one of claims 16-18, wherein the ligand-containing antisense oligonucleotide has the structure: wherein N-acetylgalactosamine is attached to the 5' end of the antisense oligonucleotide.
  20. A pharmaceutical composition comprising an antisense oligonucleotide or salt thereof according to any one of claims 1-19, and a pharmaceutically acceptable carrier.

Description

Antisense oligonucleotide targeting SPTBN1 and application thereof Technical Field The invention belongs to the field of biological medicine, and in particular relates to an antisense oligonucleotide for inhibiting SPTBN1 expression and application thereof in resisting neovascular eye diseases, hyperlipidemia and nonalcoholic fatty liver (NASH). Background SPTBN1 is the most common subtype of non-erythrocyte ghosting proteins, a cytoskeletal protein that is found in all nucleated cells and is critical for the development of many organs, such as nerves, epithelium, inner ear, osteoporosis, liver and heart. The function of SPTBN1 includes not only the establishment and maintenance of cellular architecture, but also the regulation of a variety of cellular functions such as apoptosis, cell adhesion, cell diffusion, and cell cycle regulation. In an increasing number of studies, various roles of SPTBN1 in the disease have been successively discovered, such as bone structural development and fracture healing, development and maintenance of the initial segments of central nervous system axons and langerhans joints, congenital arrhythmias, acquired and congenital heart failure, and possibly sudden cardiac death, hearing development, tumors, etc. Furthermore, recently, it has been pointed out (WO 2022/104141A 1) that the use of siRNA molecules inhibiting the expression of SPTBN1 can treat obesity, non-alcohol-related fatty liver disease, non-alcohol steatohepatitis or hepatocellular carcinoma, etc. Non-alcoholic steatohepatitis (NASH) is an inflammatory subtype of non-alcoholic fatty liver disease (NAFLD), with evidence of liver steatosis, hepatocyte injury (balloon-like lesions) and inflammation, with or without liver fibrosis. Over time, NASH may progress to cirrhosis, end-stage liver disease, or require liver transplantation. In China, nearly half of chronic liver disease patients are NAFLD. Wherein NASH comprises about 45% -50% of NAFLD. Antisense oligonucleotide (ASO) therapeutics typically consist of 15-30 chemically modified nucleotides, with the nucleotides being linked primarily by phosphorothioate linkages. ASO medicine is combined with complementary target mRNA through base complementary pairing principle after entering cells, and target mRNA is degraded under the action of ribonuclease H1, so that the effect of inhibiting target gene expression is achieved. In addition, the mechanism of action of ASO drugs includes inhibition of translation, shear regulation, increased protein translation, etc. The incidence of neovascular ocular disease increases year by year, with the increasing degree of aging of the population, the increasing number of patients has become an important blinding cause, traditional single anti-VEGF therapy has been limited, about 40-60% of patients are effective, and one third of nAMD patients have A significant vision decline after receiving 7 years of VEGF-A blocking therapy, and each patient has A central retinal atrophy. Therefore, the new target is searched, and the development of new therapeutic drugs has important clinical significance. Disclosure of Invention In view of the shortcomings of the prior art, the inventors designed antisense oligonucleotides directed against the Sptbn1 target for knockdown of Sptbn1 expression and found that antisense oligonucleotides that inhibited SPTBN1 expression were useful for the treatment of hyperlipidemia and NASH by in vivo and in vitro experiments, and more surprisingly found that such antisense oligonucleotides were also useful for the treatment of neovascular eye diseases. Based on the above, the invention provides the following technical scheme: In one aspect, the invention provides an antisense oligonucleotide that inhibits the expression of SPTBN1, said antisense oligonucleotide targeting an SPTBN1 nucleic acid. Among them, the SPTBN1 nucleic acids include, but are not limited to, transcripts (mRNAs) disclosed in NCBI, NM-003128.3 (designated herein as SEQ ID NO: 1) and NM-178313.3 (designated herein as SEQ ID NO: 2). Preferably, the antisense oligonucleotide is at least 96%, 97%, 98%, 99% complementary or fully (100%) complementary to SEQ ID NO. 1 or SEQ ID NO. 2. Preferably, the antisense oligonucleotide has 16-20 nucleotides. Further preferably, the antisense oligonucleotide has a sequence as shown in SEQ ID NO. 4-16, 18-24, 26-34, 36-43, 48-54, 56-57, 60-68, 70-78. Further preferably, the antisense oligonucleotide has the sequence shown in SEQ ID NO. 78. Preferably, at least one nucleoside in the antisense oligonucleotide comprises a modified sugar. Preferably, the modified sugar is a 2' -modified sugar. Preferably, the 2 '-modified sugar is a 2' -O-methoxyethyl modification and/or a limiting ethyl modification. Preferably, at least one nucleoside in the antisense oligonucleotide comprises a modified nucleobase. Preferably, the modified nucleobase is 5-methylcytosine. Preferably, at least one internucleoside linkage in the antisens