ASO drugs targeting hnRNPAB gene and their use in preparing drugs for treating Alzheimer's disease

Abstract

The present invention discloses a targeted hnRNPAB ASO drugs targeting genes and their use in the preparation of drugs for treating Alzheimer's disease. Specifically, the applicant identified the role of two protein subtypes of hnRNPAB in regulating the alternative splicing of AD-related genes, and the targeting hnRNPAB Genetic ASO drug screening is used for subsequent AD treatment and has prospects for clinical translation.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology and provides a targeted hnRNPAB Gene-targeting ASO drugs and their use in the preparation of drugs for treating Alzheimer's disease. Background Art

[0002] Alzheimer's disease (AD) is a major global public health challenge in the 21st century. Its core pathological features are the deposition of amyloid beta (Aβ) plaques, neurofibrillary tangles formed by abnormal phosphorylation of tau protein, and neurodegenerative lesions mediated by neuroinflammation. Among them, the pathological mechanism of tau protein is particularly critical: under normal physiological conditions, tau protein participates in key processes such as axonal transport, cell signaling, and neurogenesis by stabilizing neuronal microtubules. However, when the protein encoding tau protein MAPT Gene mutations or abnormal post-transcriptional regulation increase the tendency of tau to aggregate and interfere with its clearance, leading to the abnormal accumulation of tau protein aggregates in neurons. Abnormal tau protein aggregation is a key pathological mechanism that leads to synaptic damage, neuronal death, and ultimately cognitive decline in AD patients.

[0003] More than 90% of genes in eukaryotes produce different protein isoforms through alternative splicing, a process precisely regulated by splicing regulatory elements (SREs) and their associated splicing factors. APOER2 Alternative splicing of exon 19 of the gene (apolipoprotein E receptor 2) directly influences its function: the long isoform retained in exon 19 mediates the Reelin / Psd-95 signaling pathway, promoting synaptic plasticity and exacerbating Aβ pathology; whereas the short isoform generated by exon 19 skipping, due to the deletion of key domains, can block the cascade of Aβ toxicity signals, providing an important paradigm for targeting multiple pathological links in AD through splicing intervention. Similarly, the formation of tau pathology is closely related to the process of RNA alternative splicing. MAPT Alternative splicing of exon 10 of the gene produces 3R-tau and 4R-tau isoforms, promoting MAPT Exon 10 skipping has been found to improve synaptic function and spatial memory deficits. Specific isoforms (such as 1N4R) are more susceptible to Aβ oligomer induction and abnormal tau phosphorylation. Therefore, selectively promoting exon 10 skipping, while preserving the physiological function of normal 3R-tau, while reducing the production of pathological 4R-tau, will provide a specific intervention target for AD treatment.

[0004] Among novel therapies based on RNA splicing regulation, antisense oligonucleotides (ASOs) technology offers unique advantages. ASOs are typically short, single-stranded oligonucleotides that target pre-mRNA through Watson-Crick complementary pairing. ASOs have two mechanisms of action: one type, through DNA:RNA pairing, recruits ribonuclease H (RNase H), which cleaves RNA within the heteroduplex, thereby reducing protein expression. IONIS-MAPTRx exploits this mechanism and has demonstrated potential to reduce tau protein levels in Phase I clinical trials. However, MAPTRx acts on all tau proteins in the brain, including those that affect normal function, and is unable to precisely regulate tau protein. Therefore, there is an urgent need to find a strategy to selectively reduce abnormal tau aggregates without altering normal tau protein for the treatment of AD. Another type of ASO, which precisely modulates alternative splicing to alter the expression levels of different protein isoforms, may offer a more promising approach for AD treatment.

[0005] hnRNPAB protein belongs to the heterogeneous nuclear ribonucleoprotein (hnRNP) family of common splicing factors. SMN2 Alternative splicing of exon 7 plays a classic role in splicing regulation. The human RNA-binding protein hnRNPAB comprises three isoforms: hnRNPAB332, encoding a full-length protein of 332 amino acids; hnRNPAB285, a truncated protein of 285 amino acids resulting from alternative splicing of exon 7; and a protein isoform encoding 327 amino acids. However, the role of different hnRNPAB isoforms in the treatment of AD remains unclear. Summary of the Invention

[0006] In view of this, the object of the present invention is to provide a targeted hnRNPAB Gene-targeting ASO drugs and their use in the preparation of drugs for treating Alzheimer's disease.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] 1. Use of a hnRNPAB332 protein expression promoter in the preparation of a drug for treating Alzheimer's disease. The amino acid sequence of the hnRNPAB332 protein is shown in SEQ ID NO. 44.

[0009] 2. Use of an expression promoter of the gene encoding hnRNPAB332 protein in the preparation of a drug for treating Alzheimer's disease. The gene encoding hnRNPAB332 protein ( hnRNPABThe nucleotide sequence of the gene is shown in SEQ ID NO.47.

[0010] SEQ ID NO.47:

[0011] ATGTCGGAAGCGGGCGAGGAGCAGCCCATGGAGACGACGGGCGCCACCGAGAACGGACATGAGGCCGTCCCCGAAGGCGAGTCGCCGGCCGGGGCTGGCACGGGCGCCGCGGCGGGGGCTGGAGGCGCGACCGCGGCGCCCCCGAGCGGGAATCAGAACGGCGCCGAGGGCGACCAGATCAACGCCAGCAAGAACGAGGAGGACGCGGGAAAAATGTTCGTTGGTGGCCTGAGCTGGGATACTAGCAAAAAAGATTTAAAAGACTATTTTACTAAATTTGGAGAGGTCGTTGACTGTACAATAAAAATGGATCCCAACACTGGACGGTCAAGAGGGTTTGGGTTTATCCTGTTCAAAGATGCAGCCAGTGTGGAGAAGGTCCTAGACCAGAAGGAGCACAGGCTGGATGGCCGTGTCATTGACCCTAAAAAGGCCATGGCTATGAAGAAGGACCCGGTGAAGAAAATCTTCGTTGGGGGTCTGAATCCTGAAGCCACTGAGGAAAAGATCAGGGAGTACTTTGGCGAGTTTGGGGAGATTGAGGCCATTGAATTGCCAATGGATCCAAAGTTGAACAAAAGACGAGGTTTTGTGTTTATCACCTTTAAAGAAGAAGAACCCGTGAAGAAGGTTCTGGAGAAAAAGTTCCATACTGTCAGTGGAAGCAAGTGTGAGATCAAGGTGGCCCAGCCCAAAGAAGTCTATCAGCAGCAGCAGTATGGCTCTGGGGGCCGTGGAAACCGCAACCGAGGGAACCGAGGCAGCGGAGGTGGTGGTGGAGGTGGAGGTCAGAGTCAGAGTTGGAATCAGGGCTACGGCAACTACTGGAACCAGGGCTACGGCTACCAGCAGGGCTACGGGCCTGGCTATGGCGGCTACGACTACTCGCCCTATGGCTATTACGGCTACGGCCCCGGCTACGACTACAGTCAGGGTAGTACAAACTACGGCAAGAGCCAGCGACGTGGTGGCCATCAGAATAACTACAAGCCATACTGA。

[0012] 3. Targeting hnRNPAB ASOs of genes, including:

[0013] ASO-29: gcccgtggaccatttagcat, as shown in SEQ ID NO. 29;

[0014] ASO-30: ggaccatttagcatacgcag, as shown in SEQ ID NO. 30;

[0015] ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO. 31;

[0016] ASO-32: atacgcaggagctaggatgg, as shown in SEQ ID NO. 32;

[0017] ASO-36: gatggggcctccctctctcc, as shown in SEQ ID NO.36.

[0018] Preferably, when the final concentration of ASO is 100 nM, the ASO is selected from any one of the following:

[0019] ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO. 31;

[0020] ASO-32: atacgcaggagctaggatgg, as shown in SEQ ID NO. 32;

[0021] ASO-36: gatggggcctccctctctcc, as shown in SEQ ID NO.36.

[0022] Preferably, when the final concentration of ASO is 50 nM, the ASO is selected from any one of the following:

[0023] ASO-29: gcccgtggaccatttagcat, as shown in SEQ ID NO. 29;

[0024] ASO-30: ggaccatttagcatacgcag, as shown in SEQ ID NO. 30;

[0025] ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO.31.

[0026] More preferably, selected from any one of the following:

[0027] ASO-30: ggaccatttagcatacgcag, as shown in SEQ ID NO. 30;

[0028] ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO.31.

[0029] 4. A pharmaceutical composition comprising the aforementioned ASO.

[0030] Preferably, in the pharmaceutical composition, the final concentration of ASO is 100 nM, and the ASO is selected from any one of the following:

[0031] ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO. 31;

[0032] ASO-32: atacgcaggagctaggatgg, as shown in SEQ ID NO. 32;

[0033] ASO-36: gatggggcctccctctctcc, as shown in SEQ ID NO.36.

[0034] Preferably, in the pharmaceutical composition, the final concentration of ASO is 50 nM, and the ASO is selected from any one of the following:

[0035] ASO-29: gcccgtggaccatttagcat, as shown in SEQ ID NO. 29;

[0036] ASO-30: ggaccatttagcatacgcag, as shown in SEQ ID NO. 30;

[0037] ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO.31.

[0038] More preferably, selected from any one of the following:

[0039] ASO-30: ggaccatttagcatacgcag, as shown in SEQ ID NO. 30;

[0040] ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO.31.

[0041] 5. The aforementioned targeting hnRNPAB Use of an ASO drug or drug composition targeting a gene in the preparation of a drug for treating Alzheimer's disease.

[0042] The beneficial effects of the present invention are:

[0043] The present invention provides a targeted hnRNPAB ASO drugs targeting genes and their use in the preparation of drugs for treating Alzheimer's disease. Specifically, the applicant identified the role of two protein subtypes of hnRNPAB in regulating the alternative splicing of AD-related genes, and the targeting hnRNPAB Genetic ASO drug screening is used for subsequent AD treatment and has prospects for clinical translation.

[0044] First, the expression of hnRNPAB332 and hnRNPAB285 protein isoforms in the hippocampus of 9-month-old 3×Tg transgenic AD mouse models was detected. Full-length transcriptome sequencing was used to analyze differentially spliced ​​genes, and combined with cell transfection, the expression of the two protein isoforms in AD-related genes was identified. MAPT and ApoER2 Regulatory role of alternative splicing. Studies have found that hnRNPAB332 expression is downregulated in AD mice. hnRNPAB332 can inhibit MAPT Exon 10 alternative splicing and promotion APOER2 Exon 19 is alternatively spliced, while hnRNPAB285 does not show obvious splicing regulatory function, so by targeting hnRNPAB Exon 7 ASO drug design promotes hnRNPAB Variable splicing and upregulation of hnRNPAB332 expression will be another important strategy for treating AD.

[0045] By constructing T7-tag-hnRNPAB332 and T7-tag-hnRNPAB285 expression vectors and co-transfecting them into HeLa cells with T7 empty vector, full-length transcriptome sequencing was performed to analyze the differentially spliced ​​genes, and RT-PCR was used to identify the differentially spliced ​​genes. MAPT Exon 10 and ApoER2 Alternative splicing of exon 19 confirms the important role of hnRNPAB protein in AD treatment.

[0046] The ASO screening of the present invention adopts the ASO stepping method. hnRNPAB The 141 bp exon 7 sequence of the gene and its 62 bp upstream intron 6 sequence and 63 bp downstream intron 7 sequence were screened using a 20-base per ASO, with a step shift of 6 bases. The ASOs were modified with 2'MOE and PS. First, they were transfected into HeLa cells at a concentration of 100 nM, and the ASO regulation was detected by non-deformation polyacrylamide gel electrophoresis after RT-PCR. hnRNPAB The role of alternative splicing of exon 7. hnRNPAB ASO drugs with alternative splicing.

[0047] Other advantages, objects, and features of the present invention will be described in part in the following description and, in part, will be apparent to those skilled in the art upon examination of the following description or may be learned from practice of the present invention. The objects and other advantages of the present invention may be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be described in detail below with reference to the accompanying drawings, in which:

[0049] Figure 1 The expression of two hnRNPAB protein isoforms in AD transgenic mice, among which A is the expression of hnRNPAB332 isoform in 3×Tg mice, and B is the expression of hnRNPAB285 isoform in 3×Tg mice;

[0050] Figure 2 is the ratio of ΔPSI>0 and ΔPSI<0 among the four alternative splicing types;

[0051] Figure 3 is the ratio of the three alternative splicing types;

[0052] Figure 4 The distribution of ΔPSI of various alternative splicing types;

[0053] Figure 5 The regulatory effect of different hnRNPAB subtypes on the alternative splicing of AD-related genes is analyzed, where A represents the regulatory effect of different hnRNPAB subtypes on the alternative splicing of AD-related genes. ApoER2 Regulatory effect of alternative splicing of exon 19, B is the effect of different hnRNPAB isoforms on MAPT The regulatory role of exon 10 alternative splicing;

[0054] Figure 6 for hnRNPAB ASO screening on exon 7, where A is hnRNPAB ASO screening pattern on exon 7, B is ASO1-22 pair hnRNPAB Regulatory effect of ASO1-22 on alternative splicing of exon 7. hnRNPAB Statistical analysis of the regulatory role of exon 7 alternative splicing;

[0055] Figure 7 for hnRNPAB The partial sequence of exon 7 and the ASO screening results on intron 6 and intron 7, where A is hnRNPAB Partial sequence of exon 7 and ASO screening pattern on intron 6 and intron 7. B is ASO23-42 pair hnRNPABRegulatory effect of ASO23-42 on alternative splicing of exon 7. hnRNPAB Statistical analysis of the regulatory role of exon 7 alternative splicing. DETAILED DESCRIPTION

[0056] The present invention will be further described below in conjunction with specific embodiments.

[0057] The ASOs used in the examples of the present invention all have three modifications at the sugar site: 2′-O-methoxyethyl, phosphorothioate bond, and 5-methylcytosine (5mC) modification.

[0058] The HeLa cells involved in the embodiments of the present invention are cells disclosed in non-patent literature, available to the public, and purchased from Wuhan Punosai Biotechnology Co., Ltd.

[0059] Example 1

[0060] Differential expression of hnRNPAB332 and hnRNPAB285 in AD transgenic mice and wild-type mice

[0061] This example examined the expression of hnRNPAB332 and hnRNPAB285 in the hippocampus of 9-month-old 3×Tg transgenic mice and wild-type mice. After harvesting mouse hippocampal tissue, protein lysis buffer was added to extract total protein. Western blot analysis revealed a significant decrease in the expression of hnRNPAB332 in the hippocampus of 3×Tg transgenic mice, while an increase in hnRNPAB285 expression was observed. This suggests that upregulating hnRNPAB332 may be involved in the treatment of AD. Figure 1 shown.

[0062] Example 2

[0063] hnRNPAB332 is involved in regulating the alternative splicing of AD-related genes

[0064] (1) Construction of hnRNPAB332 / AB285 / AB327 expression vector

[0065] The amino acid sequences of hnRNPAB332, hnRNPAB327, and hnRNPAB285 described in the examples of the present invention are shown in SEQ ID NOs. 44-46. The target fragments were amplified from HeLa cell cDNA when constructing the expression vectors. The expression vectors in this section were cloned using seamless ligation near the multiple cloning site of the pCGT7 expression vector, obtained from Cold Spring Harbor Laboratory in the United States. This vector contains an 11-amino acid T7 tag (MASMTGGQQMG). Western blotting can be used to verify expression of the constructed expression vectors using a T7 primary antibody (purchased from Abcam).

[0066] The amino acid sequences of hnRNPAB332 and hnRNPAB285 are as follows:

[0067] SEQ ID NO.44 (hnRNPAB332):

[0068] MSEAGEEQPMETTGATENGHEAVPEGESPAGAGTGAAAGAGGATAAPPSGNQNGAEGDQINASKNEEDAGKMFVGGLSWDTSKKDLKDYFTKFGEVVDCTIKMD PNTGRSRGFGFILFKDAASVEKVLDQKEHRLDGRVIDPKKAMAMKKDPVKKIFVGGLNPEATEEKIREYFGEFGEIEAIELPMDPKLNKRRGFVFITFKEEEPVKKVLEKKFHT VSGSKCEIKVAQPKEVYQQQQYGSGGRGNRNRGNRGSGGGGGGGGQSQSWNQGYGNYWNQGYGYQQGYGPGYGGYDYSPYGYYGYGPGYDYSQGSTNYGKSQRRGGHQNNYKPY.

[0069] SEQ ID NO.45 (hnRNPAB327):

[0070] MSEAGEEQPMETTGATENGHEAVPEGESPAGAGTGAAAGAGGATAAPPSGNQNGAEGDQINASKNEEDAGKMFVGGLSWDTSKKDLKDYFTKFGEVVDCTIKMDPNTGRSRGFGFILFKDAASVEKVLDQKEHRLDGRVIDPKKAMAMKKDPVKKIFVGGLNPE ATEEKIREYFGEFGEIEAIELPMDPKLNKRRGFVFITFKEEEPVKKVLEKKFHTVSSGSKCEIKVAQPKEVYQQQQYGSGGRGNRNRGNRGSGGGQSQSWNQGYGNYWNQGYGYQQGYGPGYGGYDYSPYGYYGYGPGYDYSQGSTNYGKSQRRGGHQNNYKPY.

[0071] SEQ ID NO.46 (hnRNPAB285):

[0072] MSEAGEEQPMETTGATENGHEAVPEGESPAGAGTGAAAGAGGATAAPPSGNQNGAEGDQINASKNEEDAGKMFVGGLSWDTSKKDLKDYFTKFGEVVDCTIKMDPNTGRSRGFGFILFKDAASVEKVLDQKEHRLDGRVIDPK KAMAMKKDPVKKIFVGGLNPEATEEKIREYFGEFGEIEELPMDPKLNKRRGFVFITFKEEEPVKKVLEKKFHTVSSGSKCEIKVAQPKEVYQQQQYGSGGRGNRNRGNRGSGGGGGGGGQGSTNYGKSQRRGGHQNNYKPY.

[0073] (2) Full-length transcriptome sequencing to detect the alternative splicing process regulated by hnRNPAB332

[0074] The pCGT7-hnRNPAB332 expression vector and the pCGT7-hnRNPAB285 expression vector were transiently transfected into HeLa cells, and the pCGT7 empty vector was used as a control. After 48 h, the cells were collected and the whole-field transcriptome sequencing was performed to analyze the differential genes in the alternative splicing process, such as Figures 2-4Overexpression of the AB332 protein isoform is closely associated with various types of alternative splicing processes, resulting in changes in 1020 alternative splicing events, including 341 genes with exon skipping and 130 genes with intron retention. There are 360 ​​alternative splicing events with ΔPSI>0 and 660 alternative splicing events with ΔPSI<0, including classic therapeutic targets for AD. MAPT and ApoER2 Gene.

[0075] (3) Verification of the regulation of alternative splicing of AD-related genes by multiple hnRNPAB protein subtypes

[0076] The pCGT7-hnRNPAB332 or pCGT7-hnRNPAB285 expression vector was transiently transfected into HeLa cells, and RNA was extracted. Specific primers were used to amplify endogenous MAPT and ApoER2 mRNA products of gene expression, and analysis of the effects of different hnRNPAB protein isoforms on MAPT and ApoER2 Regulatory role of alternative splicing, including detection of endogenous MAPT mRNA products were detected using 30 cycles. ApoER2 The mRNA product was subjected to 28 cycles, and the RT-PCR results were as follows. Figure 5 As shown, hnRNPAB332 strongly inhibits MAPT Alternative splicing of exon 10, while promoting ApoER2 The difference between hnRNPAB332 and hnRNPAB285 is that hnRNPAB Exon 7 of the gene undergoes alternative splicing, thus enhancing hnRNPAB Alternative splicing of exon 7, selectively promoting the expression of hnRNPAB332 protein isoforms, is a new strategy for treating AD.

[0077] Example 3

[0078] ASO walking method was used to screen antisense oligonucleotides that promoted alternative splicing of hnRNPAB exon 7 and enhanced the expression of AB332.

[0079] The present invention is centered around hnRNPAB 22 ASOs were designed for screening based on the sequence of exon 7. Each ASO was 20 nt long and there was a 6 nt overlap between two adjacent ASOs. Figure 6 As shown. ASO sequences and target sequences are complementary based on the Watson-Crick principle and named according to the target location. ASOs with a final concentration of 100 nM were transfected into HeLa cells using transient transfection to verify the affinity of each ASO. hnRNPAB Effects of exon 7 inclusion were evaluated using ASO-free (Buffer) and ASO-00 (Control) as controls. Cells were harvested 30 hours later and total RNA was extracted. Fluorescence RT-PCR analysis hnRNPAB The splicing changes of exon 7 were analyzed using the following primers:

[0080] Upstream primer (hnRNPAB-F): GGTGGAGGTGGAGGTCAGAG, as shown in SEQ ID NO. 48;

[0081] Downstream primer (hnRNPAB-R): TATCCAAACAAAGCATGTGTGCG, as shown in SEQ ID NO.49.

[0082] The results showed that ASO1-22 had different degrees of splicing inhibition. Figure 6 shown.

[0083] The present invention continues to focus on hnRNPAB Twenty ASOs were designed for screening in the intron sequence flanking exon 7. Each ASO was 20 nt long, and there was a 6 nt overlap between two adjacent ASOs. Figure 7 After transfection with ASO at a final concentration of 100 nM, the results of fluorescent RT-PCR showed that multiple ASOs exhibited splicing promotion effects, among which ASO31, 32 and 36 were significant. Figure 7 shown.

[0084] Next, ASO23-42 was analyzed at a low dose by transfecting HeLa cells at a final concentration of 50 nM and analyzing the results using the above method. hnRNPAB Splicing changes of exon 7. It was found that at a concentration of 50 nM, multiple ASOs also exhibited splicing-promoting effects, among which ASO29, 30, and 31 had significant splicing-promoting effects, and the effect of ASO29 was slightly weaker than that of ASO30 and ASO31.

[0085] The ASO sequences are shown below (in order, SEQ ID NO. 1 to 43):

[0086] ASO-1:TCCAACTCTGACTCTGACct

[0087] ASO-2: CCTGATTCCAACTCTGACTC

[0088] ASO-3:CGTAGCCCTGATTCCAACTC

[0089] ASO-4:AGTTGCCGTAGCCCTGATTC

[0090] ASO-5:TCCAGTAGTTGCCGTAGCCC

[0091] ASO-6:CCTGGTTCCAGTAGTTGCCG

[0092] ASO-7:CGTAGCCCTGGTTCCAGTAG

[0093] ASO-8:GGTAGCCGTAGCCCTGGTTC

[0094] ASO-9:CCTGCTGGTAGCCGTAGCCC

[0095] ASO-10:CGTAGCCCTGCTGGTAGCCG

[0096] ASO-11:CAGGCCCGTAGCCCTGCTGG

[0097] ASO-12:CATAGCCAGGCCCGTAGCCC

[0098] ASO-13:AGCCGCCATAGCCAGGCCCG

[0099] ASO-14:AGTCGTAGCCGCCATAGCCA

[0100] ASO-15:GCGAGTAGTCGTAGCCGCCA

[0101] ASO-16:CATAGGGCGAGTAGTCGTAG

[0102] ASO-17:AATAGCCATAGGGCGAGTAG

[0103] ASO-18:AGCCGTAATAGCCATAGGGC

[0104] ASO-19:GGCCGTAGCCGTAATAGCCA

[0105] ASO-20:AGCCGGGGCCGTAGCCGTAA

[0106] ASO-21:AGTCGTAGCCGGGGCCGTAG

[0107] ASO-22:tacTGTAGTCGTAGCCGGGG

[0108] ASO-23:TCTGACTCTGACctgtgggg

[0109] ASO-24:TCTGACctgtggggggagca

[0110] ASO-25:ctgtggggggagcagggcac

[0111] ASO-26:ggggagcagggcacaggggc

[0112] ASO-27:cagggcacaggggcccgtgg

[0113] ASO-28:acaggggcccgtggaccatt

[0114] ASO-29:gcccgtggaccatttagcat

[0115] ASO-30:ggaccatttagcatacgcag

[0116] ASO-31:tttagcatacgcaggagcta

[0117] ASO-32:atacgcaggagctaggatgg

[0118] ASO-33:cctacttacTGTAGTCGTAG

[0119] ASO-34:ctctctcctacttacTGTAG

[0120] ASO-35:gcctccctctctcctactta

[0121] ASO-36:gatggggcctccctctctcc

[0122] ASO-37:tgagcggatggggcctccct

[0123] ASO-38:gaggggtgagcggatggggc

[0124] ASO-39:ggggacgaggggtgagcgga

[0125] ASO-40:tcccctggggacgaggggtg

[0126] ASO-41:cctgcctcccctggggacga

[0127] ASO-42:cactgtcctgcctcccctgg

[0128] ASO-00: CACCTTTGATACAACTACCG.

[0129] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions, which should all be included in the scope of the claims of the present invention.

Claims

1. Targeting hnRNPAB ASO of a gene, characterized in that include: ASO-29: gcccgtggaccatttagcat, as shown in SEQ ID NO. 29; ASO-30: ggaccatttagcatacgcag, as shown in SEQ ID NO. 30; ASO-31: tttagcatacgcaggagcta, as shown in SEQ ID NO. 31; ASO-32: atacgcaggagctaggatgg, as shown in SEQ ID NO. 32; ASO-36: gatggggcctccctctctcc, as shown in SEQ ID NO.

36.

2. A pharmaceutical composition comprising the ASO according to claim 1.

3. The targeting agent according to claim 1 hnRNPAB Use of an ASO targeting a gene or the pharmaceutical composition according to claim 2 in the preparation of a drug for treating Alzheimer's disease.

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