WO2022099543A1 - Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用 - Google Patents

Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用 Download PDF

Info

Publication number
WO2022099543A1
WO2022099543A1 PCT/CN2020/128367 CN2020128367W WO2022099543A1 WO 2022099543 A1 WO2022099543 A1 WO 2022099543A1 CN 2020128367 W CN2020128367 W CN 2020128367W WO 2022099543 A1 WO2022099543 A1 WO 2022099543A1
Authority
WO
WIPO (PCT)
Prior art keywords
eso
tumor
gene
application
gene inhibitor
Prior art date
Application number
PCT/CN2020/128367
Other languages
English (en)
French (fr)
Inventor
万晓春
章桂忠
刘曌
程建
逯晓旭
Original Assignee
深圳先进技术研究院
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 深圳先进技术研究院 filed Critical 深圳先进技术研究院
Priority to PCT/CN2020/128367 priority Critical patent/WO2022099543A1/zh
Publication of WO2022099543A1 publication Critical patent/WO2022099543A1/zh

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/10—Cells modified by introduction of foreign genetic material

Definitions

  • the invention belongs to the technical field of gene therapy, and relates to the application of a NY-ESO-1 gene inhibitor as a sensitizer for anti-tumor chemotherapeutics.
  • Chemotherapy is the abbreviation of chemical drug therapy. As the name suggests, it mainly uses chemical drugs to kill cancer cells to achieve the purpose of tumor treatment. Chemotherapy is one of the most effective ways to treat cancer, and together with surgery and radiotherapy, it is called the three major cancer treatments. Unlike surgery and radiotherapy, chemotherapy is a systemic treatment. No matter what route of administration is used (oral or intravenous, etc.), the drug will spread throughout most organs and tissues in the body along with the blood circulation. Therefore, chemotherapy is the main treatment method for some tumors with a tendency to spread and for intermediate and advanced tumors that have metastasized.
  • Cisplatin is the first-line chemotherapeutic drug for the treatment of various malignant tumors, and it belongs to the most commonly used and basic chemotherapeutic drugs of platinum. It has been widely used in the treatment of testicular cancer, ovarian cancer, uterine cancer, bladder cancer, neck cancer, prostate cancer, brain cancer, etc., with remarkable curative effect. Like other chemotherapeutic drugs, cisplatin is a cytotoxic drug, which causes cross-linking by binding to DNA, thereby destroying the function of DNA and inhibiting cell mitosis. It is a non-specific drug of cells. Therefore, the application process will produce large toxic and negative effects, such as nausea and vomiting, nephrotoxicity, neurotoxicity, bone marrow suppression, hair loss, etc.
  • tumor resistance often occurs during cisplatin treatment, resulting in treatment failure. Therefore, some methods that can enhance the sensitivity of tumor chemotherapy, reduce the toxic and negative effects caused by the dose, or solve the problem of clinical drug resistance are needed clinically.
  • NY-ESO-1 is a well-known cancer-testis antigen (Cancer/Testis Antigen, CTA), encoded by the CTAG1B gene. Re-expressed, including neuroblastoma, myeloma, metastatic melanoma, synovial sarcoma, bladder cancer, esophageal cancer, liver cancer, head and neck cancer, non-small cell lung cancer, ovarian cancer, prostate cancer and breast cancer.
  • the re-expression of NY-ESO-1 in tumor cells can often cause a strong immune response, with strong immunogenicity and unique expression pattern, so it is widely used in the development of tumor immunotherapy.
  • a number of tumor vaccines or TCR-T cell therapies are currently in preclinical or clinical studies. Nonetheless, the biological function of NY-ESO-1 itself is currently unclear.
  • the purpose of the present invention is to provide an application of a NY-ESO-1 gene inhibitor as a sensitizer for anti-tumor chemotherapeutics.
  • the present invention provides the use of a NY-ESO-1 gene inhibitor as a sensitizer for anti-tumor chemotherapeutics.
  • the present invention provides an application of a NY-ESO-1 gene inhibitor in preparing a drug for improving the sensitivity of tumor cells to anti-tumor chemotherapeutic drugs.
  • the present invention provides an application of a NY-ESO-1 gene inhibitor combined with an anti-tumor chemotherapeutic drug in the preparation of a tumor drug.
  • the present invention provides a method for improving the sensitivity of tumor cells to anti-tumor chemotherapeutic drugs, which is characterized by comprising the following steps: using a NY-ESO-1 gene inhibitor to knock down the expression of NY-ESO-1 in tumor cells .
  • the present invention provides a pharmaceutical composition comprising at least one antitumor chemotherapeutic drug and at least one NY-ESO-1 gene inhibitor.
  • the present invention provides a sensitizer, including NY-ESO-1 gene inhibitor.
  • the NY-ESO-1 gene inhibitor is designed according to the transcript NM_001327.3 of the gene CTAG1B encoding the human NY-ESO-1 protein.
  • the NY-ESO-1 gene inhibitor is a double-stranded shRNA that suppresses the expression of the NY-ESO-1 gene
  • the double-stranded shRNA that suppresses the expression of the NY-ESO-1 gene is such as SEQ ID NO: 1 and SEQ ID NO: 2, or SEQ ID NO: 3 and SEQ ID NO: 4.
  • anti-tumor chemotherapeutic drugs are selected from cisplatin, mitomycin and other drugs that can cause cross-linking damage between DNA strands.
  • the tumor or tumor cells are selected from solid tumors such as liver cancer, lung cancer, breast cancer, and melanoma.
  • the present invention discloses a method for enhancing the chemosensitivity of tumors to anti-tumor chemotherapeutic drugs.
  • Using shRNA to knock down the expression of NY-ESO-1 in tumor cells can effectively improve cancer cells' sensitivity to chemotherapeutic drugs such as cisplatin.
  • the sensitivity of NY-ESO-1 enhances the anti-tumor effect of such chemotherapeutic drugs, indicating that NY-ESO-1 can help tumor cells respond to the killing of chemotherapeutic drugs, and is a target for enhancing the sensitivity of tumor chemotherapy, providing a potential choice for tumor clinical treatment.
  • the disclosure of the invention provides new ideas and experimental basis for the optimization of clinical chemotherapy regimens, and has important theoretical value and potential application prospects.
  • Fig. 1 is the construction diagram of the NY-ESO-1 silent cell line in Example 3 of the present invention, wherein A is a flow-sorted GFP-positive A375 cell, and B is a fluorescent photo of a stable cell line after screening;
  • Fig. 2 is a verification diagram of the silencing effect of NY-ESO-1 in Example 3 of the present invention, wherein A is the expression of mRNA level of NY-ESO-1 in shRNA-silencing A375 cells, and B is shRNA-silencing NY-ESO-1 in A375 cells expression at the protein level;
  • Fig. 3 is a graph showing tumor cell death induced by CDDP induced by NY-ESO-1 silencing in Example 4 of the present invention, wherein the left graph is a flow chart, and the right graph is a statistical analysis result, **P ⁇ 0.01, the difference is extremely significant;
  • Figure 4 is a graph showing the in vivo anti-tumor effect of CDDP enhanced by silencing of NY-ESO-1 in Example 5 of the present invention, wherein A is a graph of tumor volume change after treatment, B is a representative graph of tumor, and C is a graph of tumor weight statistics; ns: Not significant , the difference is not significant; *P ⁇ 0.05, the difference is significant; **P ⁇ 0.01, the difference is extremely significant.
  • shRNAs Two shRNAs, shNY-1 and shNY-2, were designed for the transcript of human NY-ESO-1 protein-encoding gene CTAG1B (NM_001327.3).
  • Each shRNA includes a sense strand and an antisense strand, and its base sequence is as follows:
  • Antisense strand
  • Antisense strand
  • shNY-1 and shNY-2 were cloned into the lentiviral expression vector pLL3.7-GFP by conventional enzyme digestion and enzyme ligation methods to construct recombinant plasmids pLL3.7-GFP-shNY-1 and pLL3. 7-GFP-shNY-2.
  • recombinant plasmids to package lentivirus: Take the accurately identified lentiviral recombinant plasmids pLL3.7-GFP-shNY-1, pLL3.7-GFP-shNY-2 and control pLL3.7-GFP, respectively, and the packaging plasmids pMD2.G, PsPAX2 was co-transfected into HEK293T cells at a molar ratio of 1:1:1.
  • the transfection method refers to the operation instructions of Lipofectamine 3000 reagent.
  • the virus-containing cell supernatant was collected at 48-72h after transfection, centrifuged at 3000rpm for 10min at room temperature, and then aliquoted into 1.5mL centrifuge tubes, 0.5mL/tube, and stored at -80°C for later use or directly used for cell culture. Infect.
  • Embodiment 3 Cell infection, sorting and interference effect verification
  • Example 5 Nude mouse xenograft model to verify the effect of shRNA on the anti-tumor effect of cisplatin in vivo
  • mice with transplanted tumor size ⁇ 50mm 3 were selected and randomly divided into control group (Ctrl group) and cisplatin treatment group (CDDP group). A total of 5 injections.
  • the control group was injected with an equal volume of normal saline according to the same scheme.
  • the human shRNA involved in the present invention can efficiently silence the expression of NY-ESO-1 in tumor cells, significantly improve the sensitivity of cancer cells to cisplatin, and enhance the anti-tumor therapeutic effect of cisplatin in vitro and in vivo, and has potential clinical application value .

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Veterinary Medicine (AREA)
  • Microbiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Cell Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

一种NY-ESO-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用，NY-ESO-1基因抑制剂为抑制NY-ESO-1基因表达的双链shRNA，抑制NY-ESO-1基因表达的双链shRNA如SEQ ID NO：1和SEQ ID NO：2，或SEQ ID NO：3和SEQ ID NO：4所示。这为临床上化疗方案优化提供了新的思路和实验依据，具有重要的理论价值及潜在的应用前景。

Description

NY-ESO-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用 技术领域
本发明属于基因治疗技术领域，涉及一种NY-ESO-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用。
背景技术
尽管随着生物医药技术的发展，人们在肿瘤治疗方面的选择越来越多，但是化疗依然是肿瘤临床治疗的主要手段之一。化疗是化学药物治疗的简称，顾名思义，主要通过使用化学药物杀灭癌细胞以达到肿瘤治疗的目的。化疗是治疗癌症最有效的手段之一，和手术、放疗一起并称癌症的三大治疗手段。与手术和放疗不同，化疗是一种全身性的治疗手段，无论采用什么途径给药(口服或静脉注射等)，药物都会随着血液循环遍布全身多数器官和组织。因此，对一些有扩散倾向的肿瘤及已经转移的中晚期肿瘤，化疗都是主要的治疗手段。
顺铂是治疗多种恶性肿瘤的一线化疗药物，属于铂类最常用、最基础的化疗药物。已普遍用于治疗睾丸癌、卵巢癌、子宫癌、膀胱癌、颈部癌、前列腺癌、脑癌等，疗效显著。同其他化疗药物一样，顺铂属于细胞毒药物，通过与DNA结合引起交叉联结，从而破坏DNA的功能、抑制细胞有丝分裂，为一种细胞非特异性药物。因此，应用过程中会产生较大的毒负作用，如恶心呕吐、肾毒性、神经毒性、骨髓抑制、脱发等。此外，由于肿瘤细胞的异质性和不稳定性，顺铂治疗过程中也时常发生肿瘤耐药，导致治疗失败。因此，临床上需要一些能够增强肿瘤化疗敏感性的方法，降低剂量导致的毒负作用，或解决临床耐药问题。
NY-ESO-1是一个著名的癌-睾丸抗原(Cancer/Testis Antigen,CTA)，由CTAG1B基因编码，在正常组织中仅表达在睾丸生殖细胞和胎盘滋养细胞上，但在多种癌症类型中重新表达，包括神经母细胞瘤、骨髓瘤、转移性黑色素瘤、滑膜肉瘤、膀胱癌、食管癌、肝癌、头颈癌、非小细胞肺癌、卵巢癌、前列腺癌和乳腺癌。NY-ESO-1在肿瘤细胞中重新表达往往能够引起强烈的免疫反应，具有较强的免疫原性和独特的表达模式，因而被广泛用于肿瘤免疫治疗的开发。目前已有多项肿瘤疫苗或TCR-T细胞疗法处于临床前或临床研究中。尽管如此，NY-ESO-1本身的生物学功能目前还不清楚。
发明内容
为了解决上述背景技术中所提出的问题，本发明的目的在于提供一种NY-ESO-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用。
为达到上述目的，本发明所采用的技术方案为：
一方面，本发明提供了一种NY-ESO-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用。
另一方面，本发明提供了一种NY-ESO-1基因抑制剂在制备提高肿瘤细胞对抗肿瘤化疗药物敏感性的药物中的应用。
另一方面，本发明提供了一种NY-ESO-1基因抑制剂与抗肿瘤化疗药物联用在制备治疗肿瘤药物中的应用。
再一方面，本发明提供了一种提高肿瘤细胞对抗肿瘤化疗药物敏感性的方法，其特征在于，包括以下步骤：采用NY-ESO-1基因抑制剂敲减肿瘤细胞NY-ESO-1的表达。
再一方面，本发明提供了一种药物组合物，包含至少一种抗肿瘤化疗药物，以及至少一种NY-ESO-1基因抑制剂。
再一方面，本发明提供了一种增敏剂，包括NY-ESO-1基因抑制剂。
进一步地，所述NY-ESO-1基因抑制剂根据人NY-ESO-1蛋白的编码基因CTAG1B的转录本NM_001327.3设计。
进一步地，所述NY-ESO-1基因抑制剂为抑制NY-ESO-1基因表达的双链shRNA，所述抑制NY-ESO-1基因表达的双链shRNA如SEQ ID NO：1和SEQ ID NO：2，或SEQ ID NO：3和SEQ ID NO：4所示。
进一步地，所述抗肿瘤化疗药物选自顺铂、丝裂霉素及其他能够引起DNA链间交联损伤的药物。
进一步地，所述肿瘤或肿瘤细胞选自肝癌、肺癌、乳腺癌、黑色素瘤等实体瘤。
本发明的有益效果是：本发明公开了一种增强肿瘤对抗肿瘤化疗药物化疗敏感性的方法，利用shRNA敲减肿瘤细胞NY-ESO-1的表达，能够有效提高癌细胞对顺铂等化疗药物的敏感性，增强该类化疗药物的抗肿瘤效果，说明NY-ESO-1能够帮助肿瘤细胞应对化疗药物杀伤，是增强肿瘤化疗敏感性的靶点，为肿瘤临床治疗提供了潜在的选择。本发明的公开为临床上化疗方案优化提供了新的思路和实验依据，具有重要的理论价值及潜在的应用前景。
附图说明
图1是本发明实施例3中NY-ESO-1沉默细胞株构建图，其中A为流式分选GFP阳性的A375细胞，B为筛选后稳定细胞株的荧光照片；
图2是本发明实施例3中NY-ESO-1沉默效果验证图，其中A为shRNA沉默A375细胞中NY-ESO-1的mRNA水平的表达，B为shRNA沉默A375细胞中NY-ESO-1的蛋白水平的 表达；
图3是本发明实施例4中NY-ESO-1沉默促进CDDP诱导的肿瘤细胞死亡图，其中左图为流式图，右图为统计分析结果，**P<0.01，差异极显著；
图4是本发明实施例5中NY-ESO-1沉默增强CDDP体内抗肿瘤效果图，其中A为治疗后肿瘤体积变化图，B为肿瘤代表图，C为瘤重统计图；ns:Not significant，差异不显著；*P<0.05，差异显著；**P<0.01，差异极显著。
具体实施方式
以下通过具体实施例对本发明作详细说明，以下实施例仅用于说明本发明，而不用于限定本发明的范围。下述实施例中的试验方法，如无特殊说明，均为常规方法，所用实验材料均为市售产品。
实施例1、shRNA设计与构建
根据shRNA设计的基本原则，针对人NY-ESO-1蛋白的编码基因CTAG1B的转录本(NM_001327.3)设计2条shRNA，即shNY-1和shNY-2。每条shRNA分别包括正义链和反义链，其碱基序列如下：
shNY-1
正义链:
5’-TGCCGGCAACATACTGACTATTTCAAGAGAATAGTCAGTATGTTGCCGGCTTTTTTC-3’,SEQ ID NO:1
反义链：
5’-TCGAGAAAAAAGCCGGCAACATACTGACTATTCTCTTGAAATAGTCAGTATGTTGCCGGCA-3’,SEQ ID NO:2
shNY-2
正义链:
5’-TGCTTCAGGGCTGAATGGATTTCAAGAGAATCCATTCAGCCCTGAAGCTTTTTTC-3’,SEQ ID NO:3
反义链：
5’-TCGAGAAAAAAGCTTCAGGGCTGAATGGATTCTCTTGAAATCCATTCAGCCCTGAAGCA-3’,SEQ ID NO:4。
利用常规的酶切、酶连的方法将shNY-1和shNY-2的编码序列克隆至慢病毒表达载体pLL3.7-GFP上，以构建重组质粒pLL3.7-GFP-shNY-1和pLL3.7-GFP-shNY-2。
实施例2、慢病毒包装
采用重组质粒包装慢病毒：取鉴定准确的慢病毒重组质粒pLL3.7-GFP-shNY-1，pLL3.7-GFP-shNY-2和对照pLL3.7-GFP，分别与包装质粒pMD2.G、PsPAX2按摩尔比1:1:1的比例共转染到HEK293T细胞中。转染方法参照Lipofectamine 3000试剂的操作说明。于转染后48-72h收集含病毒的细胞上清液，室温下于3000rpm离心10min，然后分装至1.5mL离心管中，0.5mL/管，于-80℃保存备用或直接用于细胞的感染。
实施例3、细胞感染、分选及干扰效果验证
用实施例2中制备得到的含病毒的细胞上清液感染处于对数生长期的人恶性黑色素瘤细胞A375。感染后细胞继续传代培养3-5代，使病毒基因与细胞基因组完全整合。然后，利用流式细胞仪分选以获得稳定细胞株(shRNA与GFP共表达，故可用GFP作为分选依据)，分别命名为shNY-1细胞、shNY-2细胞和pLL3.7细胞，实验结果如图1所示，图1A流式分选GFP阳性的A375细胞；图1B则是筛选后稳定细胞株的荧光显微照片，从图中可以看出，筛选后的细胞均为GFP阳性，说明稳定细胞株构建成功。最后采用荧光定量PCR法和免疫印迹法分别检测胞内NY-ESO-1的mRNA和蛋白质水平，以评价所述shRNA抑制NY-ESO-1基因表达的效果，实验结果如图2所示，从图2可以看出，本发明涉及的shRNA可高效沉默A375细胞中NY-ESO-1的mRNA和蛋白水平的表达。
实施例4、流式检测shRNA对顺铂诱导的肿瘤细胞死亡的影响
(1)取处于对数生长期的shNY-1细胞、shNY-2细胞和pLL3.7细胞，0.25％胰酶消化后制成单细胞悬液；
(2)收集细胞悬液，室温下1000rpm离心3-5min，去上清；然后加适量新鲜培养基重悬，计数；
(3)以1-2×10 5cells/well铺12孔板，过夜培养后，实验组(CDDP)加入顺铂(CDDP，10μM)，对照组(Ctrl)则加等体积生理盐水；
(4)继续培养18-24h，收集细胞(包括上清中的细胞)，PBS洗涤后PI染色，流式细胞术检测细胞死亡水平(PI +％)，实验结果如图3所示，从图3可以看出对照组NY-ESO-1沉默不影响细胞死亡，而实验组中NY-ESO-1沉默明显促进细胞死亡，说明NY-ESO-1沉默促进CDDP诱导的肿瘤细胞死亡。
实施例5、裸鼠移植瘤模型验证shRNA对顺铂体内抗肿瘤作用的影响
(1)构建A375细胞皮下移植瘤裸鼠模型
取处于对数生长期的shNY-1细胞和pLL3.7细胞，0.25％胰酶消化后制成单细胞悬液，计数并调整细胞浓度为2×10 6/100μL，接种于Balb/c nude裸鼠右大腿背侧皮下，建立裸鼠移植瘤模型。
(2)顺铂对裸鼠移植瘤的实验性治疗
接种10天后，选取移植瘤大小～50mm 3小鼠，随机分为对照组(Ctrl组)和顺铂治疗组(CDDP组)，顺铂组给予腹腔注射2mg/kg的CDDP，每3天一次，共注射5次。对照组则按同样的方案注射等体积生理盐水。观察移植瘤生长情况，按公式计算瘤的体积(V＝(length x width^2)/2)，结果如图4A所示，从图中可以看出，对照组内shNY-1和pLL3.7移植瘤生长没有明显差别，而顺铂治疗组内shNY-1移植瘤明显比PLL3.7移植瘤小。第18天处死小鼠，分离移植瘤，拍照并对瘤重进行分析，实验结果如图4B和4C所示，从图中可以看出，对照组内shNY-1和PLL3.7移植瘤重量没有明显差别，而顺铂治疗组内shNY-1移植瘤的重量明显比PLL3.7移植瘤小，说明NY-ESO-1敲减不影响对照组移植瘤的重量，但是显著增强CDDP的抗肿瘤效果。
综上，本发明涉及的人shRNA能够高效沉默肿瘤细胞NY-ESO-1的表达，显著提高癌细胞对顺铂的的敏感性，增强顺铂体内外抗肿瘤治疗效果，具有潜在的临床应用价值。
以上所述仅为本发明的具体实施方式，不是全部的实施方式，本领域普通技术人员通过阅读本发明说明书而对本发明技术方案采取的任何等效的变换，均为本发明的权利要求所涵盖。

Claims (10)

  1. NY-ESO-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用。
  2. NY-ESO-1基因抑制剂在制备提高肿瘤细胞对抗肿瘤化疗药物敏感性的药物中的应用。
  3. NY-ESO-1基因抑制剂与抗肿瘤化疗药物联用在制备治疗肿瘤药物中的应用。
  4. 一种提高肿瘤细胞对抗肿瘤化疗药物敏感性的方法，其特征在于，包括以下步骤：采用NY-ESO-1基因抑制剂敲减肿瘤细胞NY-ESO-1的表达。
  5. 一种药物组合物，其特征在于，包含至少一种抗肿瘤化疗药物，以及至少一种NY-ESO-1基因抑制剂。
  6. 一种增敏剂，其特征在于，包括NY-ESO-1基因抑制剂。
  7. 根据权利要求1-3任一项所述的应用、权利要求4所述的方法、权利要求5所述的药物组合物或权利要求6所述的增敏剂，其特征在于，所述NY-ESO-1基因抑制剂根据人NY-ESO-1蛋白的编码基因CTAG1B的转录本NM_001327.3设计。
  8. 根据权利要求7所述的应用、方法、药物组合物或增敏剂，其特征在于，所述NY-ESO-1基因抑制剂为抑制NY-ESO-1基因表达的双链shRNA，所述抑制NY-ESO-1基因表达的双链shRNA如SEQ ID NO：1和SEQ ID NO：2，或SEQ ID NO：3和SEQ ID NO：4所示。
  9. 根据权利要求1-3任一项所述的应用、权利要求4所述的方法或权利要求5所述的药物组合物，其特征在于，所述抗肿瘤化疗药物选自顺铂、丝裂霉素及其他能够引起DNA链间交联损伤的药物。
  10. 根据权利要求1-3任一项所述的应用、权利要求4所述的方法或权利要求5所述的药物组合物，其特征在于，所述肿瘤或肿瘤细胞选自肝癌、肺癌、乳腺癌、黑色素瘤。
PCT/CN2020/128367 2020-11-12 2020-11-12 Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用 WO2022099543A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2020/128367 WO2022099543A1 (zh) 2020-11-12 2020-11-12 Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2020/128367 WO2022099543A1 (zh) 2020-11-12 2020-11-12 Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用

Publications (1)

Publication Number Publication Date
WO2022099543A1 true WO2022099543A1 (zh) 2022-05-19

Family

ID=81601976

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/128367 WO2022099543A1 (zh) 2020-11-12 2020-11-12 Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用

Country Status (1)

Country Link
WO (1) WO2022099543A1 (zh)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008137705A1 (en) * 2007-05-04 2008-11-13 Boris Skurkovich Prevention of cancers by immunization
CN101312741A (zh) * 2005-09-21 2008-11-26 牛津生物医学（英国）有限公司 化学免疫治疗方法
CN103483441A (zh) * 2012-06-10 2014-01-01 复旦大学 恒河猴ny-eso-1蛋白、其编码基因及应用
WO2020025642A1 (en) * 2018-08-03 2020-02-06 Ludwig Institute For Cancer Research Ltd. Viral vectors encoding cancer/testis antigens for use in a method of prevention or treatment of cancer
CN111337678A (zh) * 2020-02-21 2020-06-26 杭州凯保罗生物科技有限公司 与肿瘤免疫治疗效果相关的生物标志物及其应用
WO2020178738A1 (en) * 2019-03-04 2020-09-10 University Health Network T cell receptors and methods of use thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101312741A (zh) * 2005-09-21 2008-11-26 牛津生物医学（英国）有限公司 化学免疫治疗方法
WO2008137705A1 (en) * 2007-05-04 2008-11-13 Boris Skurkovich Prevention of cancers by immunization
CN103483441A (zh) * 2012-06-10 2014-01-01 复旦大学 恒河猴ny-eso-1蛋白、其编码基因及应用
WO2020025642A1 (en) * 2018-08-03 2020-02-06 Ludwig Institute For Cancer Research Ltd. Viral vectors encoding cancer/testis antigens for use in a method of prevention or treatment of cancer
WO2020178738A1 (en) * 2019-03-04 2020-09-10 University Health Network T cell receptors and methods of use thereof
CN111337678A (zh) * 2020-02-21 2020-06-26 杭州凯保罗生物科技有限公司 与肿瘤免疫治疗效果相关的生物标志物及其应用

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
CHÜEH ANDERLY C., LIEW MUN-SEM, RUSSELL PRUDENCE A., WALKIEWICZ MARZENA, JAYACHANDRAN APARNA, STARMANS MAUD H.W., BOUTROS PAUL C.,: "Promoter hypomethylation of NY-ESO-1, association with clinicopathological features and PD-L1 expression in non-small cell lung cancer", ONCOTARGET, vol. 8, no. 43, 26 September 2017 (2017-09-26), pages 74036 - 74048, XP055931703, DOI: 10.18632/oncotarget.18198 *
DEB S, JOHN T, CHEN Y T, AZAD A, STARMANS M H W, WALKIEWICZ M, RUSSELL P, ALTORKI N K, BOUTROS P C, MITCHELL P, CEBON J S, ZHANG X: "NY-ESO-1 AS A PREDICTIVE MARKER FOR NEOADJUVANT CHEMOTHERAPY IN STAGE 3A NON- SMALL-CELL LUNG CANCER (NSCLC) ", PATHOLOGY, vol. 44, no. s1, 1 January 2012 (2012-01-01), pages s110, XP055931696 *
LI, FEI: "Study of NY-ESO-1 as A Therapeutic Target for Multiple Myeloma", CHINESE MASTER’S THESES FULL-TEXT DATABASE, MEDICINE & HEALTH SCIENCES, 6 June 2018 (2018-06-06), pages 1 - 60, XP055931708, [retrieved on 20220615] *
MA, MING: "Advances in NY-ESO-1 Expression in Tumors and the Clinical Significance Thereof", INTERNATIONAL JOURNAL OF LABORATORY MEDICINE, vol. 36, no. 21, 1 November 2015 (2015-11-01), pages 1 - 4, XP055931706 *
PAGOTTO ANNA, CABALLERO OTAVIA L., VOLKMAR NORBERT, DEVALLE SYLVIE, SIMPSON ANDREW J. G., LU XIN, CHRISTIANSON JOHN C.: "Centrosomal Localisation of the Cancer/Testis (CT) Antigens NY-ESO-1 and MAGE-C1 Is Regulated by Proteasome Activity in Tumour Cells", PLOS ONE, vol. 8, no. 12, 1 December 2013 (2013-12-01), pages e83212, XP055931698, DOI: 10.1371/journal.pone.0083212 *
TSUJI TAKEMASA, MATSUZAKI JUNKO, CABALLERO OTAVIA L., JUNGBLUTH ACHIM A., RITTER GERD, ODUNSI KUNLE, OLD LLOYD J., GNJATIC SACHA: "Heat Shock Protein 90-Mediated Peptide-Selective Presentation of Cytosolic Tumor Antigen for Direct Recognition of Tumors by CD4 + T Cells", THE JOURNAL OF IMMUNOLOGY, WILLIAMS & WILKINS CO., US, vol. 188, no. 8, 15 April 2012 (2012-04-15), US , pages 3851 - 3858, XP055931695, ISSN: 0022-1767, DOI: 10.4049/jimmunol.1103269 *
WANG HUIYU, CHEN DATIAN, WANG RUNJIE, QUAN WEN, XIA DANDAN, MEI JIE, XU JUNYING, LIU CHAOYING: "NY-ESO-1 expression in solid tumors predicts prognosis : A systematic review and meta-analysis", MEDICINE, WILLIAMS AND WILKINS, BALTIMORE., US, vol. 98, no. 48, 1 January 2019 (2019-01-01), US , pages e17990, XP055931702, ISSN: 0025-7974, DOI: 10.1097/MD.0000000000017990 *

Similar Documents

Publication Publication Date Title
CN111315873B (zh) 分离的重组溶瘤痘病毒、药物组合物及其在治疗肿瘤和/或癌症的药物中的用途
CN111317820B (zh) 剪接因子prpf31抑制剂用于制备药物的用途
CN115772232A (zh) 一种靶向gpc3的嵌合抗原受体单核/巨噬细胞及构建方法和应用
CN110570922B (zh) 一种评估hr缺陷模型及应用
WO2017162185A1 (zh) 对非小细胞肺癌具有抑制作用的核糖核酸适配体及包含其的药物组合物
WO2022099543A1 (zh) Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用
US10302629B2 (en) Compositions and methods for treating cancer by rational targeting of protein translation
CN112316149A (zh) 一种治疗tp53突变的晚期难治性实体瘤的药物及应用
WO2022188491A1 (zh) 一种肿瘤化疗药物组合物
CN114470206A (zh) Ny-eso-1基因抑制剂作为抗肿瘤化疗药物增敏剂的应用
JP7311177B2 (ja) Ａ－ＮＯＲ－５αアンドロスタン薬物と抗がん薬物との併用
WO2022252139A1 (zh) Ny-eso-1基因抑制剂在制备抑制肿瘤转移的药物中的应用
CN111419832A (zh) 药物组合物及其在制备治疗肿瘤药物中的用途
CN113633654A (zh) 一种靶向药物及其制备方法和应用
CN108324707B (zh) 非诺贝特单药及联合化疗药在制备抗肿瘤药物中的应用
WO2019201192A1 (zh) 一种用于治疗肿瘤的柯萨奇b组病毒
Ali et al. Recent developments in targeting breast cancer stem cells (BCSCs): a descriptive review of therapeutic strategies and emerging therapies
CN110960546A (zh) MicroRNAs在制备索拉非尼治疗肝癌的增强剂中的应用
CN113402630B (zh) 一种壳聚糖衍生物递药载体及其制备方法和应用
CN115429880A (zh) Ny-eso-1基因抑制剂在制备抑制肿瘤转移的药物中的应用
CN116036094B (zh) 一种Rab13基因和EGFR蛋白表达的抑制剂及应用
CN116421590B (zh) 二乙酸氯己定在制备预防或/和治疗肝癌的药物中的用途
WO2022206805A1 (zh) 一种用于治疗肺癌的rna质粒递送系统
WO2022206778A1 (zh) 一种用于治疗肺纤维化的rna质粒递送系统
KR102212699B1 (ko) 유방암 예방 또는 치료용 조성물

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20961100

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20961100

Country of ref document: EP

Kind code of ref document: A1