WO2019129234A1 - Use of aurora kinase inhibitor and alphavirus in preparing anti-tumor drug - Google Patents

Use of aurora kinase inhibitor and alphavirus in preparing anti-tumor drug Download PDF

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WO2019129234A1
WO2019129234A1 PCT/CN2018/125015 CN2018125015W WO2019129234A1 WO 2019129234 A1 WO2019129234 A1 WO 2019129234A1 CN 2018125015 W CN2018125015 W CN 2018125015W WO 2019129234 A1 WO2019129234 A1 WO 2019129234A1
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aurora kinase
kinase inhibitor
alphavirus
tumor
barasertib
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French (fr)
Chinese (zh)
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颜光美
林园
朱文博
张海鹏
梁剑开
蔡静
龚守芳
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广州威溶特医药科技有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A61K35/768—Oncolytic viruses not provided for in groups A61K35/761 - A61K35/766
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention belongs to the field of biomedicine, and particularly relates to the application of the combination of an aurora kinase inhibitor and an alphavirus in preparing an antitumor drug.
  • Oncolytic virus is a type of replicable virus that selectively infects and kills tumor cells without damaging normal cells.
  • Oncolytic virotherapy is an innovative tumor-targeted therapeutic strategy that uses natural or genetically engineered viruses to selectively infect tumor cells and replicate in tumor cells for targeted lysis. Kills the role of tumor cells, but does not damage normal cells.
  • the M1 virus (Alphavirus M1) belongs to the genus Alphavirus, and has a good application effect in the preparation of antitumor drugs.
  • Chinese invention patent application 201410425510.3 discloses that the M1 virus can selectively cause tumor cell death without affecting normal cell survival, and has a very good application prospect in anti-tumor.
  • different tumors have different sensitivities to the M1 virus. For some tumors, when the M1 virus is administered alone, the oncolytic effect is not ideal.
  • M1 when used as an anti-tumor drug, M1 is less effective for colorectal cancer, liver cancer, bladder cancer and breast cancer than pancreatic cancer, nasopharyngeal cancer, prostate cancer and melanoma; Tumor, cervical cancer, and lung cancer are second, while gastric cancer is the least significant.
  • Aurora kinases are serine/threonine kinases whose function is a regulator of many aspects of action in mitosis and cell division. There are three related mammalian aurora kinases, Aurora A, Aurora B, and Aurora C, respectively. These kinases are overexpressed in many cancers and these kinases have now been discovered as new targets for cancer therapy.
  • Aurora A is an important mitosis regulator. They are positioned in the center body and the spindle body, respectively, and participate in the regulation of G2/M conversion, acting on the maturation and separation of the centrosome and the assembly of the spindle in the early mitosis event. Overexpression of Aurora kinase-A is an essential feature of Aurora kinase-A-induced tumorigenesis.
  • Aurora B is an important class of serine/threonine kinases responsible for regulating cell mitosis.
  • Aurora B is involved in several events such as centrosome maturation, spindle assembly and maintenance, chromosome segregation, and cytokinesis.
  • Abnormally expressed Aurora B tends to cause a large number of abnormalities in the process of mitosis.
  • Aurora B may also be involved in tumor formation, and some small molecules targeting aurora have been found to have significant tumor suppressor effects.
  • Aurora C is a chromosome passenger protein, which is concentrated in the early stage of mitosis and chromosomes, and later transferred to the middle of the spindle, and finally located in the intermediate phase during cytokinesis.
  • CN 106535940A reports that Aurora kinase inhibitors are capable of potentiating the oncolytic virus herpes simplex virus HSV1716.
  • Another object of the present invention is to provide an anti-cancer synergist capable of selectively enhancing the killing effect of alphavirus on tumor cells without affecting normal cells.
  • Another object of the present invention is to provide an application of an Aurora kinase inhibitor for the preparation of an alphavirus anti-tumor synergist.
  • Another object of the present invention is to provide an antitumor pharmaceutical composition which allows the alphavirus to exert a better antitumor effect.
  • the inventors found through research and screening that Aurora kinase inhibitors can enhance the oncolysis effect of alphavirus.
  • the Aurora kinase inhibitor is a substance that inhibits Aurora kinase activity, a substance that degrades Aurora kinase, or a genetic tool that lowers Aurora kinase levels, or a combination thereof.
  • the inventors can significantly enhance the oncolysis effect of alphavirus by inhibiting Aurora kinase.
  • the inventors used a compound that inhibits the activity of Aurora kinase, Barasertib, in combination with alphavirus to act on tumor cells. The results showed that Barasertib can synergize with alphavirus to enhance antitumor effects.
  • the present invention has found for the first time that an aurora kinase inhibitor can be used as an antitumor synergist/drug resistance reversal agent for alphavirus.
  • the present invention provides the use of an Aurora kinase inhibitor for the preparation of an alphavirus anti-tumor synergist/drug reversal agent.
  • the drug resistance reversal agent means that when some alphaviruses are used as anti-tumor drugs for treating tumors, some tumors are not sensitive to alphavirus, or these tumors are resistant to alphavirus, and in this case, Recombination of tumor resistance to the alphavirus in combination with an Aurora kinase metabolic pathway inhibitor (as a drug resistance reversal agent) in combination with alphavirus
  • the Aurora kinase inhibitor includes, but is not limited to, a compound selected from the group consisting of or a derivative thereof having apolar light kinase inhibition, or a pharmaceutically acceptable salt, solvate, tautomer, or isomer thereof:
  • a compound that inhibits the activity of an Aurora kinase protein such as Barasertib, Hesperadin or CCT137690.
  • the manner in which the compound is obtained may be selected from, but not limited to, chemical separation or synthesis by itself or from commercial sources.
  • the Aurora kinase protein inhibitor is Barasertib, and the structural formula is as shown in Formula 1:
  • the Aurora kinase protein inhibitor is Hesperadin, and the structural formula is as shown in Formula 2:
  • the Aurora kinase protein inhibitor is CCT137690, and the structural formula is as shown in Formula 3:
  • Aurora kinase inhibitors also include tools for aurora kinase gene expression inhibition tools including, but not limited to, gene interference, gene silencing, and gene editing or gene knockout.
  • the Aurora kinase gene expression inhibition tool is selected from one or more of DNA, RNA, PNA, DNA-RNA-hybrid. They can be single-stranded or double-stranded.
  • Aurora kinase inhibitors may include small inhibitory nucleic acid molecules such as short interfering RNA (siRNA), double stranded RNA (dsRNA), microRNA (miRNA), ribozyme, and small hairpin RNA (shRNA), all of which can be attenuated or Elimination of expression of Aurora kinase protein.
  • siRNA short interfering RNA
  • dsRNA double stranded RNA
  • miRNA microRNA
  • ribozyme ribozyme
  • shRNA small hairpin RNA
  • These small inhibitory nucleic acid molecules may include first and second strands that hybridize to each other to form one or more double-stranded regions, each strand having a length of about 18 to 28 nucleotides, and about 18 to 23 nucleotides. The length, or 18, 19, 20, 21, 22 nucleotides in length.
  • single strands may also comprise regions that are capable of hybridizing to each other to form a double strand, such as in a shRNA molecule.
  • These small inhibitory nucleic acid molecules may include modified nucleotides while retaining this ability to attenuate or eliminate the expression of the Aurora kinase protein.
  • Modified nucleotides can be used to improve in vitro or in vivo properties such as stability, activity and/or bioavailability.
  • These modified nucleotides may contain deoxynucleotides, 2'-methyl nucleotides, 2'-deoxy-2'-fluoronucleotides, 4'-trinucleotides, locked nucleic acid (LNA) nucleosides. Acid and/or 2'-O-methoxyethyl nucleotide and the like.
  • Small inhibitory nucleic acid molecules such as short interfering RNA (siRNA), may also contain 5'- and/or 3'-cap structures to prevent exonuclease degradation.
  • a double-stranded nucleic acid consisting of a small suppressor nucleic acid molecule contains a blunt, or overhanging, nucleotide at both ends.
  • Other nucleotides may include nucleotides that result in misalignment, bulging, cycling, or oscillating base pairs.
  • Small inhibitory nucleic acid molecules can be formulated for administration, for example, by liposome encapsulation, or by incorporation into other carriers such as biodegradable polymer hydrogels, or cyclodextrins.
  • the Aurora kinase inhibitor further comprises one or more of an antibody, an antibody functional fragment, a peptide, and a peptidomimetic.
  • the antibody may be a monoclonal antibody, a polyclonal antibody, a multivalent antibody, a multispecific antibody (eg, a bispecific antibody), and/or an antibody fragment ligated to Aurora kinase.
  • the antibody may be a chimeric antibody, a humanized antibody, a CDR-grafted antibody or a human-type antibody.
  • the antibody fragment may be, for example, Fab, Fab', F(ab')2, Fv, Fd, single-chain Fv (scFv), a disulfide-bonded FV (sdFv), or a VL, VH domain.
  • the antibody may be in a conjugated form, for example, in combination with a label, a detectable label, or a cytotoxic agent.
  • the antibody may be a homotypic IgG (eg, IgGl, IgG2, IgG3, IgG4), IgA, IgM, IgE or IgD.
  • the alphavirus is selected from one or more of the M1 virus and/or the Gaeta virus.
  • the alphavirus (M1 virus, Gaeta virus) referred to in the present invention may especially refer to the currently existing virus, but does not exclude some natural mutations that may occur or undergo mutations (natural mutations, mandatory mutations, or selectivity).
  • the alphavirus described herein includes a virus that has undergone the above changes. Preferably, the above changes do not affect the action of the alphavirus described herein.
  • the aurora kinase inhibitor is a substance (for example, a compound, or an amino acid sequence, a nucleotide sequence, etc.) or a tool capable of knocking down or affecting the expression of the Aurora kinase gene or reducing the Aurora kinase protein amount or protein activity.
  • a person skilled in the art may modify, substitute, change, etc. the inhibitory compound or the gene tool, but the aurora kinase inhibitor belonging to the present invention belongs to the above-mentioned substance, compound or tool as long as it functions as the above-mentioned inhibitory aurora kinase. Homogeneous replacement.
  • the alphavirus is the M1 virus deposited under the accession number CCTCC V201423 (as deposited with the China Center for Type Culture Collection, deposited on July 17, 2014).
  • Genbank Accession No. EF011023 records a sequence of M1.
  • the Gita virus is a virus having a homology of up to 97.8% (Wen et al. Virus Genes. 2007; 35(3): 597-603) with the M1 virus, and the two have a high identity, and the M1 virus is also somewhat
  • the literature is classified as a Gaetavirus. Both can be expected to have similar efficacy.
  • a single alphavirus strain can also be administered.
  • a variety of strains and/or types of alphaviruses can also be used.
  • the invention also provides a pharmaceutical composition for treating a tumor comprising an Aurora kinase inhibitor and an alphavirus.
  • the invention also provides a pharmaceutical kit for treating a tumor comprising an Aurora kinase inhibitor or a derivative thereof or a combination thereof, and an alphavirus.
  • the drug kit differs from the composition in that the Aurora kinase inhibitor is different from the alpha virus formulation, but is packaged separately (eg, in a pill, or in a capsule, or in a tablet or vial containing an Aurora kinase inhibitor; additional pills) , or capsules, or tablets or ampoules containing alphavirus).
  • a combination of alphavirus, aurora kinase inhibitor, and alphavirus and aurora kinase inhibitor may also contain one or more adjuvants.
  • the adjuvant refers to an ingredient which can assist the therapeutic effect of the drug in the composition of the drug.
  • the drug kit can also contain individually packaged Aurora kinase inhibitors, as well as individually packaged alphaviruses.
  • the aurora kinase inhibitor in the drug kit, and the administration of the alphavirus may be administered simultaneously or in any anterior-posterior sequence, such as administration of an aurora kinase inhibitor prior to the alphavirus, or administration of an aurora kinase inhibitor following the alphavirus, or Both are administered simultaneously.
  • the patient can be a mammal. In some embodiments, the mammal can be a human.
  • the Aurora kinase protein inhibitor includes, but is not limited to, a compound that inhibits Aurora kinase protein activity, such as Barasertib (Formula 1), Hesperadin (Formula 2), or CCT137690 (Formula 3). Or for Aurora kinase gene expression inhibition tools, including but not limited to gene interference, gene silencing, and gene editing or knockout tools.
  • the alphavirus is selected from at least one of an M1 virus and a Gata virus.
  • the laser kinase inhibitor comprises Aurora A inhibitor, Aurora B inhibitor and Aurora C inhibitor.
  • Aurora A inhibitor Aurora A inhibitor
  • Aurora B inhibitor Aurora B inhibitor
  • Aurora C inhibitor Aurora C inhibitor
  • the ratio of Barasertib, Hesperadin or CCT137690 to alphavirus may alternatively be: 0.01 to 200 mg: 10 3 to 10 9 PFU; preferably 0.1 to 200 mg: 10 4 to 10 9 PFU; further preferably 0.1. ⁇ 100mg: 10 5 ⁇ 10 9 PFU.
  • the dosage is: Barasertib, Hesperadin or CCT137690 is used in the range of 0.01 mg/kg to 200 mg/kg, while the alpha virus use titer is MOI from 10 3 to 10 9 (PFU/kg); preferably Barasertib, Hesperadin or CCT137690 is used.
  • the alpha virus uses a titer of MOI from 10 4 to 10 9 (PFU/kg); more preferably Barasertib, Hesperadin or CCT137690 is used in the range of 0.1 mg/kg to 100 mg/kg,
  • the alpha virus uses a titer of MOI from 10 5 to 10 9 (PFU/kg).
  • the tumor is a solid tumor or a hematoma.
  • the solid tumor is liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer, or gastric cancer.
  • the tumor is a tumor that is insensitive to alphavirus.
  • the tumor is a tumor that is insensitive to M1 virus.
  • the Barasertib, Hesperadin or CCT137690 provided by the present invention may be an injection, a tablet, a capsule, a patch, a kit, or the like.
  • the synergistic drug of the present invention is an injection; preferably, intravenous injection can be employed.
  • the present invention has found that Aurora kinase inhibitors, for example, Barasertib, in particular, can increase the anti-tumor effect of alphaviruses in order to increase the therapeutic effectiveness of alphaviruses as antitumor drugs.
  • Aurora kinase inhibitors for example, Barasertib
  • Cytological experiments show that the combination of M1 virus and Barasertib can significantly cause morphological lesions of tumor cells, thereby significantly enhancing the inhibition of tumor cells.
  • the present inventors have found that the combination of Barasertib and alphavirus treatment of tumor cells has a significantly better killing effect on tumor cells than the same concentration of Barasertib alone.
  • MOI 0.001
  • the tumor cell survival rate was 79.9%.
  • Selected from in the specification is connected to the selected object, and can be understood as, for example, "X is selected from: A, B, C, ..., E” or "X is selected from: A, B, C, ... and One or more of E, etc., can be understood to include X, one of A, B, C, ... E, or any combination of the two, or any combination of the plurality. It is not excluded at this time that X also includes some other categories of substances.
  • the inhibitor of the present invention may be selected from aurora kinase inhibitors which are well known in the prior art, or substances which have been found to have aurora kinase inhibition by subsequent studies.
  • Human hepatocellular carcinoma Hep3B (purchased from ATCC), M1 virus (Accession No. CCTCC V201423), high glucose DMEM medium (purchased from Corning), automatic enzyme-linked detection microplate reader.
  • MTT and intracellular succinate dehydrogenase reaction when cultured to 48h, add 20 ⁇ l (5mg/ml) of MTT to each well and continue to incubate for 4 hours. At this time, microscopically formed granular particles can be observed in living cells. Blue-purple nails crystallize.

Abstract

Disclosed is the use of an aurora kinase inhibitor and an alphavirus in preparing an anti-tumor drug. The aurora kinase inhibitor can be used in preparing an alphavirus anti-tumor synergist. Disclosed are a pharmaceutical composition containing the aurora kinase inhibitor and the alphavirus, a drug kit containing the aurora kinase inhibitor and the alphavirus, and the use of the aurora kinase inhibitor and the alphavirus in treating tumors, especially those that are insensitive to the alphavirus.

Description

极光激酶抑制剂和甲病毒在制备抗肿瘤药物的应用Application of Aurora Kinase Inhibitor and Alphavirus in Preparation of Antitumor Drugs 技术领域Technical field
本发明属于生物医药领域，具体涉及极光激酶抑制剂与甲病毒的联合在制备抗肿瘤药物中的应用。The invention belongs to the field of biomedicine, and particularly relates to the application of the combination of an aurora kinase inhibitor and an alphavirus in preparing an antitumor drug.
背景技术Background technique
溶瘤病毒(oncolytic virus)是一类选择性的感染并杀伤肿瘤细胞，而不损伤正常细胞的可复制病毒。溶瘤病毒疗法(oncolytic virotherapy)是一种创新的肿瘤靶向治疗策略，它利用天然的或经基因工程改造的病毒选择性的感染肿瘤细胞，并在肿瘤细胞中复制，达到靶向性溶解、杀伤肿瘤细胞的作用，但是对正常细胞没有损伤。Oncolytic virus is a type of replicable virus that selectively infects and kills tumor cells without damaging normal cells. Oncolytic virotherapy is an innovative tumor-targeted therapeutic strategy that uses natural or genetically engineered viruses to selectively infect tumor cells and replicate in tumor cells for targeted lysis. Kills the role of tumor cells, but does not damage normal cells.
M1病毒(Alphavirus M1)属于甲病毒属(Alphavirus)，其在制备抗肿瘤药物方面具有较好的应用效果。例如中国发明专利申请201410425510.3公开了M1病毒能选择性引起肿瘤细胞死亡而不影响正常细胞存活，其在抗肿瘤方面具有非常好的应用前景。然而，不同肿瘤对M1病毒的敏感性不一，对于某些肿瘤，M1病毒单独用药时，溶瘤作用还不够理想。例如中国发明专利申请201410425510.3所记载的，M1作为抗肿瘤药物使用时，对于结直肠癌、肝癌、膀胱癌和乳腺癌的效果不如胰腺癌、鼻咽癌、***癌和黑色素瘤明显；而胶质瘤、***、肺癌则更其次；而胃癌则最不显著。The M1 virus (Alphavirus M1) belongs to the genus Alphavirus, and has a good application effect in the preparation of antitumor drugs. For example, Chinese invention patent application 201410425510.3 discloses that the M1 virus can selectively cause tumor cell death without affecting normal cell survival, and has a very good application prospect in anti-tumor. However, different tumors have different sensitivities to the M1 virus. For some tumors, when the M1 virus is administered alone, the oncolytic effect is not ideal. For example, as described in the Chinese invention patent application 201410425510.3, when used as an anti-tumor drug, M1 is less effective for colorectal cancer, liver cancer, bladder cancer and breast cancer than pancreatic cancer, nasopharyngeal cancer, prostate cancer and melanoma; Tumor, cervical cancer, and lung cancer are second, while gastric cancer is the least significant.
筛选增加溶瘤病毒肿瘤治疗效果的化合物有望增加溶瘤病毒的抗瘤谱及抗瘤强度。发明人此前申请的专利201510990705.7中，将大黄酚及其衍生生物作为溶瘤病毒的抗瘤增效剂，二者组合可以将肿瘤细胞的存活率降低至39.6％，但其抗癌强度存在很大的进步空间，此外，这种联合应用的作用机制尚不明确。Compounds that screen to increase the therapeutic effect of oncolytic virus tumors are expected to increase the antitumor profile and antitumor strength of oncolytic viruses. In the patent application 201510990705.7 previously filed by the inventor, chrysophanol and its derivative organisms are used as antitumor synergists of oncolytic viruses, and the combination of the two can reduce the survival rate of tumor cells to 39.6%, but the anticancer strength is large. In addition, the mechanism of action of this joint application is not clear.
研究特定溶瘤病毒的增效途径并不简单。尽管已有众多已被报道的对某些 溶瘤病毒具有抗肿瘤增效作用的物质。但是，不同属的溶瘤病毒往往在增效机制上表现各异。简单的挪用难以获得理想的效果。例如，作为被证实能协同增效溶瘤棒状病毒的HDAC抑制剂(Nguye,T.L.,et al.,Chemical targeting of the innate antiviral response by histone deacetylase inhibitors renders refractory cancers sensitive to viral oncolysis.Proceedings of the national academy of sciences,2008.105(39):p.14981-14986.；Shulak,L.,et al.,Histone Deacetylase Inhibitors Potentiate Vesicular Stomatitis Virus Oncolysis in Prostate Cancer Cells by Modulating NF-kB-Dependent Autophagy.Journal of Virology,2014.88(5):p.2927-2940.；Bridle,B.W.,et al.,HDAC Inhibition Suppresses Primary Immune Responses,Enhances Secondary Immune Responses,and Abrogates Autoimmunity During Tumor Immunotherapy.Molecular therapy,2013.21(4):p.887-894.)，发明人却发现，其被用于与甲病毒联用后，却没有获得类似的增效效果。这也是溶瘤病毒增效剂开发难度大的其中一个原因。It is not straightforward to study the synergistic pathways of specific oncolytic viruses. Despite the numerous reported substances that have anti-tumor synergistic effects on certain oncolytic viruses. However, oncolytic viruses of different genera often behave differently in terms of synergistic mechanisms. Simple appropriation is difficult to achieve the desired results. For example, as a HDAC inhibitor (Nguye, TL, et al., Chemical targeting of the innate antiviral response by histone deacetylase inhibitors renders refractory cancers sensitive to viral oncolysis. Proceedings of the national academy) Of sciences, 2008.105(39): p.14981-14986.; Shulak, L., et al., Histone Deacetylase Inhibitors Potentiate Vesicular Stomatitis Virus Oncolysis in Prostate Cancer Cells by Modulating NF-kB-Dependent Autophagy. Journal of Virology, 2014.88 (5): p.2927-2940.; Bridle, BW, et al., HDAC Inhibition Suppresses Primary Immune Responses, Enhances Secondary Immune Responses, and Abrogates Autoimmunity During Tumor Immunotherapy. Molecular therapy, 2013.21(4): p.887- 894.), the inventors found that it was used in combination with alphavirus, but did not achieve similar synergistic effects. This is one of the reasons why the development of oncolytic virus synergists is difficult.
极光激酶是丝氨酸/苏氨酸激酶，其功能是作用在有丝***和细胞***中的多方面的调节剂。有三种相关的哺乳动物极光激酶，分别是极光激酶-A(Aurora A)、极光激酶-B(Aurora B)和极光激酶-C(Aurora C)。这些激酶在许多癌症中过度表达，目前这些激酶已经被人发现作为癌症治疗的新靶点。Aurora kinases are serine/threonine kinases whose function is a regulator of many aspects of action in mitosis and cell division. There are three related mammalian aurora kinases, Aurora A, Aurora B, and Aurora C, respectively. These kinases are overexpressed in many cancers and these kinases have now been discovered as new targets for cancer therapy.
其中，极光激酶-A(Aurora A)是一个重要的有丝***调节因子。分别先后定位于中心体和纺锤体极，参与G2/M转换的调节，在早期有丝***事件中作用于中心体的成熟与分离及纺锤体的组装等。极光激酶-A的过度表达是极光激酶-A诱导肿瘤发生的必要特征。Among them, Aurora A is an important mitosis regulator. They are positioned in the center body and the spindle body, respectively, and participate in the regulation of G2/M conversion, acting on the maturation and separation of the centrosome and the assembly of the spindle in the early mitosis event. Overexpression of Aurora kinase-A is an essential feature of Aurora kinase-A-induced tumorigenesis.
其中，极光激酶-B(Aurora B)是负责调控细胞有丝***的一类重要的丝氨酸/苏氨酸激酶。近年来，随着Aurora B相关研究的不断深入，人们逐渐认识到Aurora B在细胞有丝***以及肿瘤形成中的重要作用。在细胞有丝***中，Aurora B参与了诸如中心体成熟分离、纺锤体组装和维持、染色体分离以及胞质***等多个事件。异常表达的Aurora B往往会导致细胞在有丝***的过程中出现大量的异常现象。此外，Aurora B还可能参与了肿瘤形成，已经发现一些 靶向作用于极光的小分子具有显著的抑癌作用。Among them, Aurora B is an important class of serine/threonine kinases responsible for regulating cell mitosis. In recent years, with the deepening of Aurora B related research, people have gradually recognized the important role of Aurora B in cell mitosis and tumor formation. In cell mitosis, Aurora B is involved in several events such as centrosome maturation, spindle assembly and maintenance, chromosome segregation, and cytokinesis. Abnormally expressed Aurora B tends to cause a large number of abnormalities in the process of mitosis. In addition, Aurora B may also be involved in tumor formation, and some small molecules targeting aurora have been found to have significant tumor suppressor effects.
其中，极光激酶-C(Aurora C)是染色体过客蛋白(chromosome passenger protein),它在有丝***早期与染色体集合，后期则转移到纺锤体中间区，最后在胞质***期定位于中间体。Among them, Aurora C is a chromosome passenger protein, which is concentrated in the early stage of mitosis and chromosomes, and later transferred to the middle of the spindle, and finally located in the intermediate phase during cytokinesis.
CN 106535940A中报道了极光激酶抑制剂能够增效溶瘤病毒单纯疱疹病毒HSV1716。CN 106535940A reports that Aurora kinase inhibitors are capable of potentiating the oncolytic virus herpes simplex virus HSV1716.
发明内容Summary of the invention
本发明的目的在于提供一种甲病毒抗肿瘤增效剂。It is an object of the present invention to provide an alphavirus anti-tumor synergist.
本发明的另一个目的在于提供能够选择性的增强甲病毒对肿瘤细胞的杀伤作用，而不影响正常细胞的抗癌增效剂。Another object of the present invention is to provide an anti-cancer synergist capable of selectively enhancing the killing effect of alphavirus on tumor cells without affecting normal cells.
本发明的另一个目的在于提供一种极光激酶抑制剂在制备甲病毒抗瘤增效剂方面的应用。Another object of the present invention is to provide an application of an Aurora kinase inhibitor for the preparation of an alphavirus anti-tumor synergist.
本发明的另一个目的在于提供一种抗瘤药物组合物，其可以使得甲病毒发挥更好的抗瘤效果。Another object of the present invention is to provide an antitumor pharmaceutical composition which allows the alphavirus to exert a better antitumor effect.
本发明的进一步目的在于提供一种针对甲病毒不敏感的肿瘤，安全有效的甲病毒增效药物。It is a further object of the present invention to provide a safe and effective alpha virus potentiating drug against a tumor that is insensitive to alphavirus.
发明通过以下技术方案实现上述目的：The invention achieves the above objects by the following technical solutions:
发明人通过研究、筛选发现，极光激酶抑制剂可以增强甲病毒的溶瘤效果。The inventors found through research and screening that Aurora kinase inhibitors can enhance the oncolysis effect of alphavirus.
所述的极光激酶抑制剂为抑制极光激酶活性的物质、或降解极光激酶的物质、或降低极光激酶水平的基因工具、或它们的组合。The Aurora kinase inhibitor is a substance that inhibits Aurora kinase activity, a substance that degrades Aurora kinase, or a genetic tool that lowers Aurora kinase levels, or a combination thereof.
发明人通过抑制极光激酶可以显著增强甲病毒的溶瘤效应。发明人采用了抑制极光激酶活性的化合物Barasertib协同甲病毒作用于肿瘤细胞，实验结果发现，Barasertib可以协同及甲病毒增强抗肿瘤效应。The inventors can significantly enhance the oncolysis effect of alphavirus by inhibiting Aurora kinase. The inventors used a compound that inhibits the activity of Aurora kinase, Barasertib, in combination with alphavirus to act on tumor cells. The results showed that Barasertib can synergize with alphavirus to enhance antitumor effects.
本发明首次发现，极光激酶抑制剂可以作为甲病毒的抗瘤增效剂/耐药逆转剂。The present invention has found for the first time that an aurora kinase inhibitor can be used as an antitumor synergist/drug resistance reversal agent for alphavirus.
本发明提供了极光激酶抑制剂在制备甲病毒抗瘤增效剂/耐药逆转剂方面的应用。The present invention provides the use of an Aurora kinase inhibitor for the preparation of an alphavirus anti-tumor synergist/drug reversal agent.
耐药逆转剂是指，当采用一些甲病毒作为抗肿瘤药物用于***时，存在着一些肿瘤对甲病毒并不太敏感，或者说这些肿瘤对甲病毒具有抗性，此时，可以采用与极光激酶代谢通路抑制剂(作为耐药逆转剂)联用甲病毒的方式，以逆转肿瘤对所述甲病毒的抗性The drug resistance reversal agent means that when some alphaviruses are used as anti-tumor drugs for treating tumors, some tumors are not sensitive to alphavirus, or these tumors are resistant to alphavirus, and in this case, Recombination of tumor resistance to the alphavirus in combination with an Aurora kinase metabolic pathway inhibitor (as a drug resistance reversal agent) in combination with alphavirus
所述的极光激酶抑制剂包括但不限于选自以下化合物或其具有极光激酶抑制作用的衍生物、或其药学上可接受的盐、溶剂化物、互变异构体、同分异构体：Barasertib、Hesperadin或CCT137690等抑制极光激酶蛋白活性的化合物。化合物的获取方式可选但不限于：自己化学分离或合成或者从商业途径购买。The Aurora kinase inhibitor includes, but is not limited to, a compound selected from the group consisting of or a derivative thereof having apolar light kinase inhibition, or a pharmaceutically acceptable salt, solvate, tautomer, or isomer thereof: A compound that inhibits the activity of an Aurora kinase protein, such as Barasertib, Hesperadin or CCT137690. The manner in which the compound is obtained may be selected from, but not limited to, chemical separation or synthesis by itself or from commercial sources.
在本发明一优选的实施例中，极光激酶蛋白抑制剂为Barasertib，其结构式如式1所示：In a preferred embodiment of the invention, the Aurora kinase protein inhibitor is Barasertib, and the structural formula is as shown in Formula 1:
Figure PCTCN2018125015-appb-000001
Figure PCTCN2018125015-appb-000001
在本发明另一优选的实施例中，极光激酶蛋白抑制剂为Hesperadin，其结构式如式2所示：In another preferred embodiment of the present invention, the Aurora kinase protein inhibitor is Hesperadin, and the structural formula is as shown in Formula 2:
Figure PCTCN2018125015-appb-000002
Figure PCTCN2018125015-appb-000002
在本发明另一优选的实施例中，极光激酶蛋白抑制剂为CCT137690，其结构式如式3所示：In another preferred embodiment of the present invention, the Aurora kinase protein inhibitor is CCT137690, and the structural formula is as shown in Formula 3:
Figure PCTCN2018125015-appb-000003
Figure PCTCN2018125015-appb-000003
在本发明一些优选的实施例中，极光激酶抑制剂还包括针对极光激酶基因表达抑制工具，包括但不限于基因干扰、基因沉默、以及基因编辑或基因敲除等工具手段。In some preferred embodiments of the invention, Aurora kinase inhibitors also include tools for aurora kinase gene expression inhibition tools including, but not limited to, gene interference, gene silencing, and gene editing or gene knockout.
作为一种可选的实施方式，所述极光激酶基因表达抑制工具选自DNA、RNA、PNA、DNA-RNA-杂合体中的一种或几种。它们可以是单链的或双链的。As an alternative embodiment, the Aurora kinase gene expression inhibition tool is selected from one or more of DNA, RNA, PNA, DNA-RNA-hybrid. They can be single-stranded or double-stranded.
极光激酶抑制剂可包括一些小的抑制核酸分子，例如短干扰RNA(siRNA)，双链RNA(dsRNA)，microRNA(miRNA)，核酶，以及小发夹RNA(shRNA)，这些都能减弱或消除极光激酶蛋白的表达。Aurora kinase inhibitors may include small inhibitory nucleic acid molecules such as short interfering RNA (siRNA), double stranded RNA (dsRNA), microRNA (miRNA), ribozyme, and small hairpin RNA (shRNA), all of which can be attenuated or Elimination of expression of Aurora kinase protein.
这些小的抑制核酸分子可能包括第一、第二链，二者杂交彼此形成一个或多个双链区，每条链大约18～28个核苷酸的长度，大约18～23个核苷酸的长度，或者18，19，20，21，22个核苷酸的长度。另外，单链也可能包含能够相互杂交形成双链的区域，例如在shRNA分子中。These small inhibitory nucleic acid molecules may include first and second strands that hybridize to each other to form one or more double-stranded regions, each strand having a length of about 18 to 28 nucleotides, and about 18 to 23 nucleotides. The length, or 18, 19, 20, 21, 22 nucleotides in length. In addition, single strands may also comprise regions that are capable of hybridizing to each other to form a double strand, such as in a shRNA molecule.
这些小的抑制核酸分子在保持这种减弱或消除极光激酶蛋白的表达的能力时，可能包括修饰性核苷酸。修饰性核苷酸可用于改善体外或体内特性，如稳定性、活性和/或生物利用度。这些修饰性核苷酸可能含有脱氧核苷酸、2’-甲基核苷酸、2’-脱氧-2’-氟核苷酸、4’-三核苷酸、锁核酸(LNA)核苷酸和/或2’-O-甲氧乙基核苷酸等。小的抑制核酸分子，如短干扰RNA(siRNA)，也可能含有5’-和/或3’-帽结构，以此来防止核酸外切酶对其降解。These small inhibitory nucleic acid molecules may include modified nucleotides while retaining this ability to attenuate or eliminate the expression of the Aurora kinase protein. Modified nucleotides can be used to improve in vitro or in vivo properties such as stability, activity and/or bioavailability. These modified nucleotides may contain deoxynucleotides, 2'-methyl nucleotides, 2'-deoxy-2'-fluoronucleotides, 4'-trinucleotides, locked nucleic acid (LNA) nucleosides. Acid and/or 2'-O-methoxyethyl nucleotide and the like. Small inhibitory nucleic acid molecules, such as short interfering RNA (siRNA), may also contain 5'- and/or 3'-cap structures to prevent exonuclease degradation.
在一些实施例中，小抑制核酸分子组成的双链核酸含有两端钝、或悬垂的核苷酸。其他核苷酸可能包括会导致错位、凸起、循环、或摆动碱基对的核苷酸。小抑制核酸分子可以设计配方以便施用，例如，通过脂质体包裹，或掺入其他载体(如可生物降解聚合物水凝胶，或环糊精)。In some embodiments, a double-stranded nucleic acid consisting of a small suppressor nucleic acid molecule contains a blunt, or overhanging, nucleotide at both ends. Other nucleotides may include nucleotides that result in misalignment, bulging, cycling, or oscillating base pairs. Small inhibitory nucleic acid molecules can be formulated for administration, for example, by liposome encapsulation, or by incorporation into other carriers such as biodegradable polymer hydrogels, or cyclodextrins.
在本发明另一些优选的实施例中，所述的极光激酶抑制剂还包括抗体、抗体功能性片段、肽类、和拟肽类中的一种或几种。其中，所述的抗体可能是单克隆抗体，多克隆抗体，多价抗体，多特异性抗体(例如：双特异性抗体)，和/或连接在极光激酶上的抗体片段。该抗体可以是嵌合抗体、人源化抗体、CDR移植抗体或人型抗体。抗体片段可以是，例如，Fab，Fab’，F(ab’)2，Fv，Fd，单链Fv(scFv)，具二硫键的FV(sdFv)，或VL、VH结构域。抗体可能是一个共轭的形式，例如，结合一个标签、一个可检测标记，或一种细胞毒性剂。抗体可能是同型IgG(例如：IgG1、IgG2、IgG3、IgG4)、IgA、IgM、IgE或IgD。In still other preferred embodiments of the invention, the Aurora kinase inhibitor further comprises one or more of an antibody, an antibody functional fragment, a peptide, and a peptidomimetic. Wherein, the antibody may be a monoclonal antibody, a polyclonal antibody, a multivalent antibody, a multispecific antibody (eg, a bispecific antibody), and/or an antibody fragment ligated to Aurora kinase. The antibody may be a chimeric antibody, a humanized antibody, a CDR-grafted antibody or a human-type antibody. The antibody fragment may be, for example, Fab, Fab', F(ab')2, Fv, Fd, single-chain Fv (scFv), a disulfide-bonded FV (sdFv), or a VL, VH domain. The antibody may be in a conjugated form, for example, in combination with a label, a detectable label, or a cytotoxic agent. The antibody may be a homotypic IgG (eg, IgGl, IgG2, IgG3, IgG4), IgA, IgM, IgE or IgD.
所述的甲病毒病毒选自M1病毒和/或盖塔病毒中的一种或多种。The alphavirus is selected from one or more of the M1 virus and/or the Gaeta virus.
本发明所说的甲病毒(M1病毒、盖塔病毒)可以尤其地指目前已有的病毒，但也不排除一些可能发生的自然变异或者进行了突变(自然突变、强制性突变、 或选择性突变)、基因修饰、序列增加或删除或部分替换的病毒。这里所述的甲病毒包括已经进行了上述改变的病毒。最好是上述改变并不影响所说的甲病毒发挥本发明所述的作用。所说的极光激酶抑制剂为能起到敲低或影响极光激酶基因表达或者降低极光激酶蛋白量或蛋白活性的物质(例如化合物、或氨基酸序列、核苷酸序列等)或工具等。本领域技术人员可以对其抑制化合物或者基因工具进行修饰、替换、改变等，但只要起到上述抑制极光激酶作用的，则属于本发明的极光激酶抑制剂，属于上述物质、化合物或工具等的同质替换。The alphavirus (M1 virus, Gaeta virus) referred to in the present invention may especially refer to the currently existing virus, but does not exclude some natural mutations that may occur or undergo mutations (natural mutations, mandatory mutations, or selectivity). A mutation, a genetic modification, a sequence increase or deletion or partial replacement of a virus. The alphavirus described herein includes a virus that has undergone the above changes. Preferably, the above changes do not affect the action of the alphavirus described herein. The aurora kinase inhibitor is a substance (for example, a compound, or an amino acid sequence, a nucleotide sequence, etc.) or a tool capable of knocking down or affecting the expression of the Aurora kinase gene or reducing the Aurora kinase protein amount or protein activity. A person skilled in the art may modify, substitute, change, etc. the inhibitory compound or the gene tool, but the aurora kinase inhibitor belonging to the present invention belongs to the above-mentioned substance, compound or tool as long as it functions as the above-mentioned inhibitory aurora kinase. Homogeneous replacement.
在一些实施例中，甲病毒是保藏编号CCTCC V201423(保藏于中国典型培养物保藏中心，保藏日期2014年7月17日)的M1病毒。作为很可能来源于同一毒株的病毒，Genbank Accession No.EF011023记录了一株M1的序列。盖塔病毒作为与M1病毒具有高达97.8％(Wen et al.Virus Genes.2007；35(3):597-603)同源性的病毒，两者具有很高的同一性，M1病毒也被一些文献归类为类盖塔病毒。可以预期二者具有相似的功效。In some embodiments, the alphavirus is the M1 virus deposited under the accession number CCTCC V201423 (as deposited with the China Center for Type Culture Collection, deposited on July 17, 2014). As a virus most likely derived from the same strain, Genbank Accession No. EF011023 records a sequence of M1. The Gita virus is a virus having a homology of up to 97.8% (Wen et al. Virus Genes. 2007; 35(3): 597-603) with the M1 virus, and the two have a high identity, and the M1 virus is also somewhat The literature is classified as a Gaetavirus. Both can be expected to have similar efficacy.
单个甲病毒株也可以施用。在其他实施方案中，也可使用多种菌株和/或类型的甲病毒。A single alphavirus strain can also be administered. In other embodiments, a variety of strains and/or types of alphaviruses can also be used.
本发明还提供一种用于***的药物组合物，其包含极光激酶抑制剂以及甲病毒。本发明还提供用于***的药品套装，其包含极光激酶抑制剂或其衍生物或它们的组合，以及甲病毒。药品套装区别于组合物的地方在于，极光激酶抑制剂不同于甲病毒的剂型，而是独立包装(例如：药丸、或胶囊、或药片或安剖瓶中，含有极光激酶抑制剂；另外的药丸、或胶囊、或药片或安剖瓶中，含有甲病毒)。在一些实施例中，甲病毒、极光激酶抑制剂，以及甲病毒和极光激酶抑制剂的组合，也可含一种或多种佐剂。所述的佐剂是指在药物组成中，可辅助药物疗效的成分。药品套装也可以包含独立包装的极光激酶抑制剂，以及独立包装的甲病毒。药物套装中极光激酶抑制剂，以及甲病毒的施用，可以是同时施用或者是以任意的前后顺序施用，例如在甲病毒之前施用极光激酶抑制剂，或者在甲病毒之后施用极光激酶抑制剂，或者两者同时施用。在各 种实施例中，患者可以是哺乳动物。在一些实施例中，哺乳动物可以是人。The invention also provides a pharmaceutical composition for treating a tumor comprising an Aurora kinase inhibitor and an alphavirus. The invention also provides a pharmaceutical kit for treating a tumor comprising an Aurora kinase inhibitor or a derivative thereof or a combination thereof, and an alphavirus. The drug kit differs from the composition in that the Aurora kinase inhibitor is different from the alpha virus formulation, but is packaged separately (eg, in a pill, or in a capsule, or in a tablet or vial containing an Aurora kinase inhibitor; additional pills) , or capsules, or tablets or ampoules containing alphavirus). In some embodiments, a combination of alphavirus, aurora kinase inhibitor, and alphavirus and aurora kinase inhibitor may also contain one or more adjuvants. The adjuvant refers to an ingredient which can assist the therapeutic effect of the drug in the composition of the drug. The drug kit can also contain individually packaged Aurora kinase inhibitors, as well as individually packaged alphaviruses. The aurora kinase inhibitor in the drug kit, and the administration of the alphavirus, may be administered simultaneously or in any anterior-posterior sequence, such as administration of an aurora kinase inhibitor prior to the alphavirus, or administration of an aurora kinase inhibitor following the alphavirus, or Both are administered simultaneously. In various embodiments, the patient can be a mammal. In some embodiments, the mammal can be a human.
所述的极光激酶蛋白抑制剂包括但不限于Barasertib(式1)、Hesperadin(式2)或CCT137690(式3)这一类的抑制极光激酶蛋白活性的化合物。或者针对极光激酶基因表达抑制工具，包括但不限于基因干扰、基因沉默以及基因编辑或敲除等工具手段。The Aurora kinase protein inhibitor includes, but is not limited to, a compound that inhibits Aurora kinase protein activity, such as Barasertib (Formula 1), Hesperadin (Formula 2), or CCT137690 (Formula 3). Or for Aurora kinase gene expression inhibition tools, including but not limited to gene interference, gene silencing, and gene editing or knockout tools.
所述的甲病毒选自M1病毒和盖塔病毒的至少一种。The alphavirus is selected from at least one of an M1 virus and a Gata virus.
需要说明的是，本发明中，所述的激光激酶抑制剂包括极光激酶-A(Aurora A)抑制剂、极光激酶-B(Aurora B)抑制剂和极光激酶-C(Aurora C)抑制剂中的一种或几种。It should be noted that, in the present invention, the laser kinase inhibitor comprises Aurora A inhibitor, Aurora B inhibitor and Aurora C inhibitor. One or several.
在组合物或药品套装中，Barasertib、Hesperadin或CCT137690与甲病毒的配比可选地为：0.01～200mg:10 3～10 9PFU；优选0.1～200mg:10 4～10 9PFU；进一步优选0.1～100mg:10 5～10 9PFU。 In the composition or the pharmaceutical kit, the ratio of Barasertib, Hesperadin or CCT137690 to alphavirus may alternatively be: 0.01 to 200 mg: 10 3 to 10 9 PFU; preferably 0.1 to 200 mg: 10 4 to 10 9 PFU; further preferably 0.1. ~100mg: 10 5 ~ 10 9 PFU.
优选使用剂量为：Barasertib、Hesperadin或CCT137690使用范围为0.01mg/kg至200mg/kg，同时甲病毒使用滴度为MOI从10 3至10 9(PFU/kg)；优选Barasertib、Hesperadin或CCT137690使用范围为0.1mg/kg至200mg/kg，同时甲病毒使用滴度为MOI从10 4至10 9(PFU/kg)；更优选Barasertib、Hesperadin或CCT137690使用范围为0.1mg/kg至100mg/kg，同时甲病毒使用滴度为MOI从10 5至10 9(PFU/kg)。 Preferably, the dosage is: Barasertib, Hesperadin or CCT137690 is used in the range of 0.01 mg/kg to 200 mg/kg, while the alpha virus use titer is MOI from 10 3 to 10 9 (PFU/kg); preferably Barasertib, Hesperadin or CCT137690 is used. From 0.1 mg/kg to 200 mg/kg, while the alpha virus uses a titer of MOI from 10 4 to 10 9 (PFU/kg); more preferably Barasertib, Hesperadin or CCT137690 is used in the range of 0.1 mg/kg to 100 mg/kg, The alpha virus uses a titer of MOI from 10 5 to 10 9 (PFU/kg).
在一个实施方式中，所述肿瘤为实体瘤或血液瘤。在一个实施方式中，所述实体瘤为肝癌、结直肠癌、膀胱癌、乳腺癌、***、***癌、胶质瘤、黑色素瘤、胰腺癌、鼻咽癌、肺癌、或胃癌。在优选的实施方式中，所述肿瘤为对甲病毒不敏感的肿瘤。在更优选的实施方式中，所述肿瘤为对M1病毒不敏感的肿瘤。In one embodiment, the tumor is a solid tumor or a hematoma. In one embodiment, the solid tumor is liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer, or gastric cancer. In a preferred embodiment, the tumor is a tumor that is insensitive to alphavirus. In a more preferred embodiment, the tumor is a tumor that is insensitive to M1 virus.
作为可选的实施方案，本发明所提供的Barasertib、Hesperadin或CCT137690可以是注射剂、片剂、胶囊、贴剂、试剂盒等。作为优选的实施方案，本发明的增效药物是注射剂；优选地，可采用静脉注射。As an alternative embodiment, the Barasertib, Hesperadin or CCT137690 provided by the present invention may be an injection, a tablet, a capsule, a patch, a kit, or the like. As a preferred embodiment, the synergistic drug of the present invention is an injection; preferably, intravenous injection can be employed.
作为本发明进一步优选的实施方案：As a further preferred embodiment of the invention:
本发明发现了极光激酶抑制剂，例如、尤其是Barasertib可以增加甲病毒的抗肿瘤效应，以提高甲病毒作为抗肿瘤药物时的治疗有效性。细胞学实验证明M1病毒和Barasertib联合应用，可显著引起肿瘤细胞的形态学病变，从而显著增强对肿瘤细胞的抑制作用。The present invention has found that Aurora kinase inhibitors, for example, Barasertib, in particular, can increase the anti-tumor effect of alphaviruses in order to increase the therapeutic effectiveness of alphaviruses as antitumor drugs. Cytological experiments show that the combination of M1 virus and Barasertib can significantly cause morphological lesions of tumor cells, thereby significantly enhancing the inhibition of tumor cells.
我们联合Barasertib和M1病毒作用于人肝细胞癌Hep3B株，出人意料的发现Barasertib和M1病毒联合应用时，显著增加肿瘤细胞形态病变，显著降低肿瘤细胞生存率。例如在本发明的一个实施例中，当M1病毒(MOI＝0.001)单独处理肝癌细胞时，肿瘤细胞存活率为79.9％，而当以0.4μM的Barasertib与同样MOI的M1病毒联用时，肿瘤细胞存活率大幅下降至33.2％。与单用M1病毒的抗肿瘤效果相比，Barasertib与M1联用时，溶瘤效果显著提升。We combined Barasertib and M1 against human hepatocellular carcinoma Hep3B strain, and unexpectedly found that when Barasertib and M1 virus were combined, it significantly increased tumor cell morphology and significantly reduced tumor cell survival. For example, in one embodiment of the present invention, when the M1 virus (MOI=0.001) treats the liver cancer cells alone, the tumor cell survival rate is 79.9%, and when the 0.4 μM Barasertib is combined with the M1 virus of the same MOI, the tumor cells are used. The survival rate dropped significantly to 33.2%. Compared with the anti-tumor effect of the M1 virus alone, the oncolytic effect was significantly improved when Barasertib was combined with M1.
发明人此前将大黄酚及其衍生物作为M1病毒的抗癌增效剂，经试验发现，50μM的大黄酚与(MOI＝0.001)M1病毒联用后，肿瘤细胞的存活率下降至39.6％，而本发明发现，将0.4μM的Barasertib与M1病毒联用后，肿瘤细胞的存活率显著下降至33.2％。与大黄酚及其衍生物相比，本发明的M1抗肿瘤增效剂显著提高了肿瘤的杀伤率。并且，在Barasertib联合M1病毒实现了上述的杀伤率时，其所需要的药物有效剂量上仅为大黄酚的不足百分之一，具备显著优越性。The inventors previously used chrysophanol and its derivatives as anti-cancer synergists for M1 virus. It was found by experiments that when 50 μM of chrysophanol was combined with (MOI=0.001) M1 virus, the survival rate of tumor cells decreased to 39.6%. The present inventors have found that the tumor cell survival rate is significantly reduced to 33.2% after the combination of 0.4 μM of Barasertib and the M1 virus. Compared with chrysophanol and its derivatives, the M1 anti-tumor synergist of the present invention significantly increases the tumor killing rate. Moreover, when the Barasertib combined with the M1 virus achieves the above-mentioned killing rate, the effective dose of the drug required is only less than one percent of the chrysophanol, and has remarkable advantages.
本发明发现，Barasertib与甲病毒联合应用处理肿瘤细胞，对肿瘤细胞杀伤作用显著优于单用相同浓度的Barasertib，例如当同样例如以0.4μM的Barasertib处理肿瘤细胞时，肿瘤细胞存活率仍高达84.3％；M1病毒(MOI＝0.001)单独处理肝癌细胞时，肿瘤细胞存活率为79.9％。而当以0.4μM的Barasertib与M1(MOI＝0.001)病毒联用时，肿瘤细胞存活率大幅下降至33.2％。可见，Barasertib与M1联用时大幅提升的溶瘤效果，是得益于Barasertib与M1病毒之间的协同性机制，并非简单地通过Barasertib的抗肿瘤机制发挥作用。The present inventors have found that the combination of Barasertib and alphavirus treatment of tumor cells has a significantly better killing effect on tumor cells than the same concentration of Barasertib alone. For example, when tumor cells are treated with, for example, Barasertib at 0.4 μM, the tumor cell survival rate is still as high as 84.3. %; M1 virus (MOI = 0.001) When treating liver cancer cells alone, the tumor cell survival rate was 79.9%. When the 0.4 μM Barasertib was combined with the M1 (MOI=0.001) virus, the tumor cell survival rate dropped significantly to 33.2%. It can be seen that the greatly enhanced oncolytic effect of Barasertib in combination with M1 is due to the synergistic mechanism between Barasertib and M1 virus, not simply through the anti-tumor mechanism of Barasertib.
附图说明DRAWINGS
图1Barasertib与M1病毒联合处理显著降低人肝细胞癌株Hep3B生存率。Figure 1 Barasertib combined with M1 virus significantly reduced the survival rate of human hepatocellular carcinoma Hep3B.
具体实施方式Detailed ways
以下实施方式是对本发明作进一步说明，但本发明的实施方式不局限于以下的实施例介绍，凡依照本发明的原理或理念所作的等同的变化或变通都应视为本发明保护的范畴。The invention is further described in the following embodiments, but the embodiments of the present invention are not limited to the following embodiments, and equivalent changes or modifications made in accordance with the principles and concepts of the present invention are considered to be within the scope of the present invention.
在没有特别指明的情况下，本发明采用的材料及实验方法为常规材料及方法。The materials and experimental methods employed in the present invention are conventional materials and methods, unless otherwise specified.
说明书中的“选自”连接着所选对象，可以理解为，例如：“X选自：A、B、C、……、E”或“X选自：A、B、C、……和E中的一种或多种”，等等，均可理解为，X包括了A、B、C、……E中的一种、或者两者的任意组合、或者多者的任意组合。此时不排除X还包括了一些其他类别的物质。"Selected from" in the specification is connected to the selected object, and can be understood as, for example, "X is selected from: A, B, C, ..., E" or "X is selected from: A, B, C, ... and One or more of E, etc., can be understood to include X, one of A, B, C, ... E, or any combination of the two, or any combination of the plurality. It is not excluded at this time that X also includes some other categories of substances.
除了上述提及的极光激酶抑制剂，本发明的抑制剂还可以选自现有技术中已经公知的极光激酶抑制剂、或者经后续研究发现具备极光激酶抑制作用的物质。In addition to the above-mentioned aurora kinase inhibitors, the inhibitor of the present invention may be selected from aurora kinase inhibitors which are well known in the prior art, or substances which have been found to have aurora kinase inhibition by subsequent studies.
实施例1 Barasertib与M1病毒联合处理显著降低人肝细胞癌株Hep3B生存率Example 1 Barasertib combined with M1 virus significantly reduced the survival rate of human hepatocellular carcinoma Hep3B
材料：material:
人肝细胞癌Hep3B(购于ATCC)，M1病毒(保藏编号CCTCC V201423)，高糖DMEM培养基(购于Corning)，自动酶联检测酶标仪。Human hepatocellular carcinoma Hep3B (purchased from ATCC), M1 virus (Accession No. CCTCC V201423), high glucose DMEM medium (purchased from Corning), automatic enzyme-linked detection microplate reader.
方法：method:
a)接种细胞、给药处理：选择对数生长期细胞，DMEM完全培养液(含10％胎牛血清、1％双抗)制成细胞悬液，以每孔4×10 3/孔的密度接种在96孔培养板内。12小时后见细胞完全贴壁，实验分对照组，单独Barasertib组，M1感染组和Barasertib/M1联用组。所用剂量为：所用剂量为：M1病毒(MOI＝0.001) 感染细胞；Barasertib为0.4μM。 a) Inoculation of cells, administration: Select logarithmic growth phase cells, DMEM complete medium (containing 10% fetal bovine serum, 1% double antibody) to make cell suspension, with a density of 4 × 10 3 /well per well Inoculate in a 96-well culture plate. After 12 hours, the cells were fully adherent, and the experiment was divided into control group, Barasertib group, M1 infection group and Barasertib/M1 combination group. The doses used were: M1 virus (MOI = 0.001) infected cells; Barasertib 0.4 μM.
b)MTT与细胞内的琥珀酸脱氢酶反应：培养至48h时，每孔加入MTT 20μl(5mg/ml)，继续孵育4小时，此时镜检可观察到、活细胞内形成的颗粒状蓝紫色甲臜结晶。b) MTT and intracellular succinate dehydrogenase reaction: when cultured to 48h, add 20μl (5mg/ml) of MTT to each well and continue to incubate for 4 hours. At this time, microscopically formed granular particles can be observed in living cells. Blue-purple nails crystallize.
c)溶解甲臜颗粒：小心吸去上清，加DMSO 100μl/孔溶解形成的结晶，在微型振荡器上震荡5min，然后在酶联检测仪上用波长570nm检测各孔的光密度(OD值)。细胞存活率＝药物处理组OD值/对照组OD值×100％。c) Dissolving the formazan granules: carefully aspirate the supernatant, add DMSO 100 μl/well to dissolve the crystals, shake on a micro-vibrator for 5 min, and then measure the optical density (OD value) of each well with a wavelength of 570 nm on an enzyme-linked detector. ). Cell viability = OD value of the drug-treated group / OD value of the control group × 100%.
结果:result:
如图1所示，M1病毒(MOI＝0.001)单独处理对肿瘤细胞Hep3B具有较小的生存率抑制作用，肿瘤细胞存活率达到79.9％，0.4μM的Barasertib处理组肿瘤细胞存活率仍高达84.3％，然而，当同样的0.4μM的Barasertib与M1病毒(MOI＝0.001)联用(Barasertib+M1)时，肿瘤细胞存活率大幅下降至33.2％。As shown in Figure 1, M1 virus (MOI=0.001) treatment alone had a lower survival rate inhibition effect on tumor cell Hep3B, tumor cell survival rate reached 79.9%, and the survival rate of tumor cells in the 0.4μM Barasertib treatment group was still as high as 84.3%. However, when the same 0.4 μM Barasertib was combined with the M1 virus (MOI=0.001) (Barasertib+M1), the tumor cell survival rate dropped significantly to 33.2%.

Claims (10)

  1. 极光激酶抑制剂在制备甲病毒抗肿瘤增效剂或耐药逆转剂方面的应用；。The use of an Aurora kinase inhibitor in the preparation of an alphavirus anti-tumor synergist or a drug resistance reversal agent;
  2. 根据权利要求1所述的应用，其特征在于，所述的甲病毒选自M1病毒和盖塔病毒中的至少一种。The use according to claim 1, wherein the alphavirus is selected from at least one of an M1 virus and a Gata virus.
  3. 如权利要求1或2任一所述的应用，其特征在于，所述的极光激酶抑制剂为抑制极光激酶活性的物质、或降解极光激酶的物质、降低极光激酶水平的基因工具、或它们的任意组合；The use according to any one of claims 1 or 2, wherein the Aurora kinase inhibitor is a substance which inhibits aurora kinase activity, a substance which degrades Aurora kinase, a gene tool which lowers Aurora kinase level, or a random combination;
    优选地，所述的极光激酶抑制剂选自化合物；Preferably, the Aurora kinase inhibitor is selected from the group consisting of compounds;
    更优选地，所述的极光激酶抑制剂选自以下化合物或其具有极光激酶抑制作用的衍生物、或其药学上可接受的盐、溶剂化物、互变异构体、同分异构体：Barasertib、Hesperadin或CCT137690；More preferably, the Aurora kinase inhibitor is selected from the group consisting of a compound thereof or a derivative thereof having apolar light kinase inhibition, or a pharmaceutically acceptable salt, solvate, tautomer, isomer thereof: Barasertib, Hesperadin or CCT137690;
    更优选地，所述的Barasertib的结构式如式1所示：More preferably, the structural formula of the Barasertib is as shown in Equation 1:
    Figure PCTCN2018125015-appb-100001
    Figure PCTCN2018125015-appb-100001
    更优选地，所述的Hesperadin的结构式如式2所示：More preferably, the structural formula of the Hesperadin is as shown in Formula 2:
    Figure PCTCN2018125015-appb-100002
    Figure PCTCN2018125015-appb-100002
    更优选地，所述的CCT137690的结构式如式3所示：More preferably, the structural formula of the CCT 137690 is as shown in Equation 3:
    Figure PCTCN2018125015-appb-100003
    Figure PCTCN2018125015-appb-100003
    或者优选地，所述的极光激酶抑制剂选自基因干扰、基因编辑、基因沉默或基因敲除材料；Or preferably, said Aurora kinase inhibitor is selected from the group consisting of gene interference, gene editing, gene silencing or gene knockout materials;
    或者优选地，所述的极光激酶抑制剂选自DNA、RNA、PNA、DNA-RNA-杂合体中的一种或几种；Or preferably, said Aurora kinase inhibitor is selected from one or more of DNA, RNA, PNA, DNA-RNA-hybrid;
    更优选地，所述的极光激酶抑制剂选自siRNA、dsRNA、miRNA、shRNA、核酶中的一种或几种；More preferably, the Aurora kinase inhibitor is selected from one or more of siRNA, dsRNA, miRNA, shRNA, ribozyme;
    更优选地，所述的极光激酶抑制剂为肿瘤靶向的极光激酶抑制剂。More preferably, the Aurora kinase inhibitor is a tumor-targeted Aurora kinase inhibitor.
  4. 一种***的药物组合物，包含：A pharmaceutical composition for treating a tumor comprising:
    (a)极光激酶抑制剂；(a) an aurora kinase inhibitor;
    优选地，所述的极光激酶抑制剂为抑制极光激酶活性的物质、或降解极光激 酶的物质、或降低极光激酶水平的基因工具；Preferably, the Aurora kinase inhibitor is a substance that inhibits Aurora kinase activity, or a substance that degrades aurora kinase, or a genetic tool that lowers Aurora kinase levels;
    优选地，所述的极光激酶抑制剂选自化合物；Preferably, the Aurora kinase inhibitor is selected from the group consisting of compounds;
    更优选地，所述的极光激酶抑制剂选自以下化合物或其具有极光激酶抑制作用的衍生物、或其药学上可接受的盐、溶剂化物、互变异构体、同分异构体：Barasertib、Hesperadin或CCT137690；More preferably, the Aurora kinase inhibitor is selected from the group consisting of a compound thereof or a derivative thereof having apolar light kinase inhibition, or a pharmaceutically acceptable salt, solvate, tautomer, isomer thereof: Barasertib, Hesperadin or CCT137690;
    更优选地，所述的Barasertib的结构式如式1所示：More preferably, the structural formula of the Barasertib is as shown in Equation 1:
    Figure PCTCN2018125015-appb-100004
    Figure PCTCN2018125015-appb-100004
    更优选地，所述的Hesperadin的结构式如式2所示：More preferably, the structural formula of the Hesperadin is as shown in Formula 2:
    Figure PCTCN2018125015-appb-100005
    Figure PCTCN2018125015-appb-100005
    更优选地，所述的CCT137690的结构式如式3所示：More preferably, the structural formula of the CCT 137690 is as shown in Equation 3:
    Figure PCTCN2018125015-appb-100006
    Figure PCTCN2018125015-appb-100006
    或者优选地，所述的极光激酶抑制剂选自基因干扰、基因编辑、基因沉默或基因敲除材料；Or preferably, said Aurora kinase inhibitor is selected from the group consisting of gene interference, gene editing, gene silencing or gene knockout materials;
    或者优选地，所述的极光激酶抑制剂选自DNA、RNA、PNA或DNA-RNA-杂合体；Or preferably, said Aurora kinase inhibitor is selected from the group consisting of DNA, RNA, PNA or DNA-RNA-hybrid;
    更优选地，所述的极光激酶抑制剂选自siRNA、dsRNA、miRNA、shRNA或核酶；More preferably, the Aurora kinase inhibitor is selected from the group consisting of siRNA, dsRNA, miRNA, shRNA or ribozyme;
    更优选地，所述的极光激酶抑制剂为肿瘤靶向的极光激酶抑制剂。More preferably, the Aurora kinase inhibitor is a tumor-targeted Aurora kinase inhibitor.
    (b)甲病毒；优选地，所述的甲病毒选自M1病毒和盖塔病毒中的至少一种；优选地，所述药物组合物还包含药学上可接受的载体；(b) an alphavirus; preferably, the alphavirus is selected from at least one of an M1 virus and a Gatavirus; preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier;
    更优选地，所述载体优选地选自冻干粉针、注射剂、片剂、胶囊、试剂盒或贴剂。More preferably, the carrier is preferably selected from the group consisting of lyophilized powders, injections, tablets, capsules, kits or patches.
  5. 一种药品套装，包含：A medicine kit containing:
    (a)极光激酶抑制剂；(a) an aurora kinase inhibitor;
    所述的极光激酶抑制剂为抑制极光激酶活性的物质、或降解极光激酶的物质、或降低极光激酶水平的基因工具、或它们的任意组合；The Aurora kinase inhibitor is a substance that inhibits Aurora kinase activity, or a substance that degrades Aurora kinase, or a gene tool that lowers Aurora kinase level, or any combination thereof;
    优选地，所述的极光激酶抑制剂选自化合物；Preferably, the Aurora kinase inhibitor is selected from the group consisting of compounds;
    更优选地，所述的极光激酶抑制剂选自以下化合物或其具有极光激酶抑制作用的衍生物、或其药学上可接受的盐、溶剂化物、互变异构体、同分异构体： Barasertib、Hesperadin或CCT137690；More preferably, the Aurora kinase inhibitor is selected from the group consisting of a compound thereof or a derivative thereof having apolar light kinase inhibition, or a pharmaceutically acceptable salt, solvate, tautomer, isomer thereof: Barasertib, Hesperadin or CCT137690;
    更优选地，所述的Barasertib的结构式如式1所示：More preferably, the structural formula of the Barasertib is as shown in Equation 1:
    Figure PCTCN2018125015-appb-100007
    Figure PCTCN2018125015-appb-100007
    更优选地，所述的Hesperadin的结构式如式2所示：More preferably, the structural formula of the Hesperadin is as shown in Formula 2:
    Figure PCTCN2018125015-appb-100008
    Figure PCTCN2018125015-appb-100008
    更优选地，所述的CCT137690的结构式如式3所示：More preferably, the structural formula of the CCT 137690 is as shown in Equation 3:
    Figure PCTCN2018125015-appb-100009
    Figure PCTCN2018125015-appb-100009
    或者优选地，所述的极光激酶抑制剂选自基因干扰、基因编辑、基因沉默或基因敲除材料；Or preferably, said Aurora kinase inhibitor is selected from the group consisting of gene interference, gene editing, gene silencing or gene knockout materials;
    或者优选地，所述的极光激酶抑制剂选自DNA、RNA、PNA、DNA-RNA-杂合体中的一种或多种；Or preferably, said Aurora kinase inhibitor is selected from one or more of DNA, RNA, PNA, DNA-RNA-hybrid;
    更优选地，所述的极光激酶抑制剂选自siRNA、dsRNA、miRNA、shRNA、核酶中的一种或多种；More preferably, the Aurora kinase inhibitor is selected from one or more of siRNA, dsRNA, miRNA, shRNA, ribozyme;
    更优选地，所述的极光激酶抑制剂为肿瘤靶向极光激酶抑制剂；More preferably, the Aurora kinase inhibitor is a tumor-targeted Aurora kinase inhibitor;
    (b)甲病毒；(b) alphavirus;
    优选地，所述的甲病毒选自M1病毒和盖塔病毒中的至少一种；Preferably, the alphavirus is selected from at least one of an M1 virus and a Gata virus;
    优选地，在所述的药品套装中，极光激酶抑制剂和甲病毒被分开包装。Preferably, in the kit of drugs, the Aurora kinase inhibitor and the alphavirus are packaged separately.
  6. 极光激酶抑制剂及甲病毒的组合在制备***药物中的应用；The use of a combination of an Aurora kinase inhibitor and an alphavirus in the preparation of a medicament for treating tumors;
    优选地，所述的甲病毒选自M1病毒和盖塔病毒中的至少一种；Preferably, the alphavirus is selected from at least one of an M1 virus and a Gata virus;
    所述的极光激酶抑制剂为抑制极光激酶蛋白活性的物质、或降解极光激酶蛋白的物质、或降低极光激酶蛋白水平的基因工具、或它们的任意组合；The Aurora kinase inhibitor is a substance which inhibits the activity of the Aurora kinase protein, or a substance which degrades the Aurora kinase protein, or a gene tool which lowers the level of the Aurora kinase protein, or any combination thereof;
    优选地，所述的极光激酶抑制剂选自化合物；Preferably, the Aurora kinase inhibitor is selected from the group consisting of compounds;
    更优选地，所述的极光激酶抑制剂选自以下化合物或其具有极光激酶抑制作用的衍生物、或其药学上可接受的盐、溶剂化物、互变异构体、同分异构体：Barasertib、Hesperadin或CCT137690；More preferably, the Aurora kinase inhibitor is selected from the group consisting of a compound thereof or a derivative thereof having apolar light kinase inhibition, or a pharmaceutically acceptable salt, solvate, tautomer, isomer thereof: Barasertib, Hesperadin or CCT137690;
    更优选地，所述的Barasertib的结构式如式1所示：More preferably, the structural formula of the Barasertib is as shown in Equation 1:
    Figure PCTCN2018125015-appb-100010
    Figure PCTCN2018125015-appb-100010
    更优选地，所述的Hesperadin的结构式如式2所示：More preferably, the structural formula of the Hesperadin is as shown in Formula 2:
    Figure PCTCN2018125015-appb-100011
    Figure PCTCN2018125015-appb-100011
    更优选地，所述的CCT137690的结构式如式3所示：More preferably, the structural formula of the CCT 137690 is as shown in Equation 3:
    Figure PCTCN2018125015-appb-100012
    Figure PCTCN2018125015-appb-100012
    或者优选地，所述的极光激酶抑制剂为基因干扰、基因编辑、基因沉默或基因敲除材料；Or preferably, said Aurora kinase inhibitor is a gene interference, gene editing, gene silencing or gene knockout material;
    或者优选地，所述的极光激酶抑制剂选自DNA、RNA、PNA、DNA-RNA-杂合体中的一种或多种；Or preferably, said Aurora kinase inhibitor is selected from one or more of DNA, RNA, PNA, DNA-RNA-hybrid;
    更优选地，所述的极光激酶抑制剂选自siRNA、dsRNA、miRNA、shRNA、核酶中的一种或多种；More preferably, the Aurora kinase inhibitor is selected from one or more of siRNA, dsRNA, miRNA, shRNA, ribozyme;
    更优选地，所述的极光激酶抑制剂为肿瘤靶向的极光激酶抑制剂。More preferably, the Aurora kinase inhibitor is a tumor-targeted Aurora kinase inhibitor.
  7. 如权利要求4或5任一所述的组合物/药品套装，其特征在于所述的Barasertib、Hesperadin或CCT137690与甲病毒的配比为：0.01～200mg:10 3～10 9PFU；优 选0.1～200mg:10 4～10 9PFU；进一步优选0.1～100mg:10 5～10 9PFU； The composition/pharmaceutical kit according to any one of claims 4 or 5, wherein the ratio of the Barasertib, Hesperadin or CCT137690 to the alphavirus is 0.01 to 200 mg: 10 3 to 10 9 PFU; preferably 0.1 to 200 mg: 10 4 to 10 9 PFU; further preferably 0.1 to 100 mg: 10 5 to 10 9 PFU;
    进一步优选地，使用剂量为：Barasertib、Hesperadin或CCT137690使用范围为0.01mg/kg至200mg/kg，同时甲病毒使用滴度为MOI从10 3至10 9(PFU/kg)； Further preferably, the dosage is: Barasertib, Hesperadin or CCT137690 is used in the range of 0.01 mg/kg to 200 mg/kg, while the alpha virus using titer is MOI from 10 3 to 10 9 (PFU/kg);
    优选Barasertib、Hesperadin或CCT137690使用范围为0.1mg/kg至200mg/kg，同时甲病毒使用滴度为MOI从10 4至10 9(PFU/kg)；更优选Barasertib、Hesperadin或CCT137690使用范围为0.1mg/kg至100mg/kg，同时甲病毒使用滴度为MOI从10 5至10 9(PFU/kg)。 Preferably Barasertib, Hesperadin or CCT137690 is used in the range of 0.1 mg/kg to 200 mg/kg, while the alpha virus uses a titer of MOI from 10 4 to 10 9 (PFU/kg); more preferably Barasertib, Hesperadin or CCT137690 is used in a range of 0.1 mg. /kg to 100 mg/kg, while the alpha virus uses a titer of MOI from 10 5 to 10 9 (PFU/kg).
  8. 如权利要求1-7任一所述的应用/组合物/药品套装，其特征在于所述的极光激酶蛋白抑制剂为Barasertib、Hesperadin或CCT137690中的一种或几种。The use/composition/drug set according to any one of claims 1 to 7, characterized in that the Aurora kinase inhibitor is one or more of Barasertib, Hesperadin or CCT137690.
  9. 如权利要求1-8任一所述的应用/组合物/药品套装，其特征在于所述的肿瘤为实体瘤或血液瘤；优选地，所述的实体瘤为肝癌、结直肠癌、膀胱癌、乳腺癌、***、***癌、胶质瘤、黑色素瘤、胰腺癌、鼻咽癌、肺癌或胃癌；或者优选地，所述的肿瘤为对甲病毒不敏感的肿瘤；The use/composition/drug set according to any one of claims 1 to 8, wherein the tumor is a solid tumor or a hematoma; preferably, the solid tumor is liver cancer, colorectal cancer, bladder cancer Breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal carcinoma, lung cancer or gastric cancer; or preferably, the tumor is a tumor that is insensitive to alphavirus;
    更优选地，所述肿瘤为对甲病毒不敏感的肝癌、结直肠癌、膀胱癌、乳腺癌、***、***癌、胶质瘤、黑色素瘤、胰腺癌、鼻咽癌、肺癌或胃癌。More preferably, the tumor is liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer or gastric cancer which are insensitive to alphavirus.
  10. 根据权利要求1-9任一所述的应用/组合物/药品套装，其中，所述的极光激酶抑制剂选自极光激酶-A抑制剂、极光激酶-B抑制剂和极光激酶-C抑制剂中的一种或几种。The use/composition/drug set according to any one of claims 1 to 9, wherein said Aurora kinase inhibitor is selected from the group consisting of an Aurora kinase-A inhibitor, an Aurora kinase-B inhibitor, and an Aurora kinase-C inhibitor One or several of them.
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