Application of IU1 in preparation of drugs for treating p53 defective tumors
阅读说明：本技术 IU1在制备治疗p53缺陷型肿瘤的药物中的应用 (Application of IU1 in preparation of drugs for treating p53 defective tumors ) 是由 符达 马雨水 于 2019-11-15 设计创作，主要内容包括：本发明提供了IU1在制备治疗p53缺陷型肿瘤的药物中的应用,属于肿瘤药物技术领域。具有如式I所示的结构的IU1在制备治疗p53缺陷型肿瘤的药物中的应用。应用小分子抑制剂IU1对去泛素酶USP14的特异性抑制导致p53缺陷小鼠体内淋巴瘤和肉瘤的持久性肿瘤消退,而不影响正常组织,治疗反应与COPS5泛素增加有关。通过USP14的COPS5去泛素和p53依赖和独立调节机制,IU1特异性抑制USP14导致了持久的肿瘤消退。(The invention provides application of IU1 in preparation of a drug for treating p53 defective tumors, and belongs to the technical field of tumor drugs. Application of IU1 with a structure shown as a formula I in preparation of drugs for treating p53 deficient tumors. Specific inhibition of deubiquitinase USP14 with the small molecule inhibitor IU1 resulted in persistent tumor regression of lymphomas and sarcomas in p53 deficient mice without affecting normal tissues, and the therapeutic response was associated with increased COPS5 ubiquitin. Specific inhibition of USP14 by IU1 results in sustained tumor regression through COPS5 deubiquitinating and p53 dependent and independent regulatory mechanisms of USP 14.)
1. Application of IU1 with a structure shown as a formula I in preparation of drugs for treating p53 deficient tumors.
2. The use according to claim 1, characterized in that IU1 is for the purpose of treating p53 deficient tumors by specifically inhibiting the deubiquitinating activity of USP 14.
Technical Field
The invention belongs to the technical field of tumor drugs, and particularly relates to application of IU1 in preparation of a drug for treating p53 defective tumors.
Background
Oncogenes, which are a mutant form of normal genes (also called proto-oncogenes) that control cell growth and division, can cause normal cells to become cancerous, and cancer suppressor genes play an important role in the development of cancer. The proteins encoded by oncogenes mainly include several major types, such as growth factors, growth factor receptors, molecules in signal transduction pathways, gene transcription regulators, and cell cycle regulatory proteins. Cancer suppressor genes are negative regulators of normal cell proliferation and encode proteins that often act to arrest cycle progression at cell cycle checkpoints. The development of cancer is the result of accumulation of genetic mutations. In 1979, David Lane and Lionel Crawford were studying an oncogenic virus:
Normally, regulation of the cell cycle initiation p53 protein will monitor the "physical condition" of the cell. When cellular DNA is damaged (DDR), p53 protein initiates a cell cycle arrest (cell arrest) state, which in turn induces cell senescence or apoptosis. When cells have abnormal mitosis, such as amplification of centrosomes or telomere dysfunction, p53 can eliminate these abnormal cells without leaving a feeling, so as to limit the generation of chromosome instability. p53 also controls many "non-canonical" pathways such as autophagy activity, altered metabolism and cellular plasticity. In addition to inducing cell cycle arrest, senescence and apoptosis in response to acute DNA damage, p53 also regulates other aspects of cell behavior. p53 can maintain genomic stability. Based on the role of p53 in acute DNA damage response, p53 has been called "genomic guardian". In addition to this role, recent studies have shown that p53 can maintain genome integrity through other mechanisms. First, p53 transactivates various DNA repair genes and directly controls different forms of DNA repair, including mismatch repair, base excision repair, and nucleotide excision repair. In addition p53 has also been described as a daemon of the epigenome. p53 inhibits DNA methyltransferases Dnmt3a and 3b and activates Tetl and Tet2, promoting DNA demethylation. p53 inhibits glycolysis by inhibiting genes such as Glut1 and Glut4 glucose transporters, and by activating Sco2 and the like to promote mitochondrial oxidative phosphorylation. During tumor suppression, p53 does respond to some type of DNA damage. p53 may also activate tumor suppressive effects through extracellular microenvironment stress such as hypoxia and nutritional starvation.
Because the gene coding the protein p53 has multiple functions of controlling cell cycle, apoptosis, controlling neovascularization and the like, p53 becomes an attractive drug target in cancer research. There are two main aspects of drug research on p53 gene mutation: directly acting on mutant p53 gene (restoring wild-type activity of p53 and inducing degradation of mutant p53 protein) and indirectly acting on mutant p53 protein.
Because the p53 gene has strong ability to inhibit cell growth and initiate apoptosis, people can think whether to copy a wild p53 gene to the p53 defective tumor cell, thereby achieving the purpose of treating tumor. Roth et al, 993, proposed a clinical protocol for the treatment of non-small cell lung cancer using p53 or the like. Since more and more reports on p53 used in various tumor studies, the overall conclusion is that: the p53 gene therapy has the characteristic of less toxic and side effects. The effect of controlling tumor growth generated after the cancer suppressor gene leads human tumor cells is easily observed on a cellular level. Animal tumor experiments for restoring the function of the defective p53 by using wild p53 have more reports, for example, the survival period of animals is obviously prolonged by using in-situ cancer and metastatic cancer for introducing p53 into the lung by using liposome as a carrier, and the apoptosis of tumor cells is induced by using virus as a carrier to express the wild p53 in a p53 mutant mouse. However, the conventional method cannot introduce cancer suppressor genes into most cells in tumors grown in vivo, and thus it is difficult to obtain a good clinical effect.
Disclosure of Invention
In view of the above, the present invention aims to develop a pharmacological strategy for recovering p53 function and an application thereof in targeted therapy of p53 deficient mice, and therefore provides an application of a small molecule inhibitor IU1 in preparation of a drug for treating p53 deficient tumors.
The invention provides application of IU1 with a structure shown as a formula I in preparation of a drug for treating p53 defective tumors.
Preferably, said IU1 achieves the purpose of treating p53 deficient tumors by specifically inhibiting the deubiquitinating activity of USP 14.
The invention provides application of IU1 with a structure shown in formula I in preparation of a drug for treating p53 defective tumors. Specific inhibition of deubiquitinase USP14 with the small molecule inhibitor IU1 resulted in persistent tumor regression of lymphomas and sarcomas in p53 deficient mice without affecting normal tissues, and the therapeutic response was associated with increased COPS5 ubiquitin. Inhibition of USP14 resulted in sustained tumor regression through COPS5 deubiquitinating and p53 dependent and independent regulatory mechanisms of USP 14.
Drawings
FIG. 1 is a graph showing the results of tumor persistence degeneration in mice deficient in p53 gene by IU1 treatment, and FIG. 1-A is a KAPLAN-MEIER survival assay for evaluating the effect of IU1 on wild-type, heterozygous and homozygous mouse OS; FIGS. 1-B-1-E show the effect of IU1 on the whole body (FIG. 1-B), tumor detection time (FIG. 1-C), and tumor volume (FIGS. 1-D and 1-E) of wild type, p53 heterozygous knockout mice, and p53 homozygous knockout mice; CTRL, control; MOCK, untreated mouse; ND, not detected; WT, wild type; data shown are mean ± SDS. Statistical analysis was performed using one-way anova (P <0.05 and P <0.01 compared to control);
FIG. 2 shows X-ray, MICRO-CT and MRI analyses and typing of primary tumors of p53 gene homozygous defective mice, FIG. 2-A shows X-ray, MICRO-CT and MRI analyses of MLT of p53 gene homozygous defective mice, and FIG. 2-B shows X-ray, MICRO-CT and MRI analyses of STS of p53 gene homozygous defective mice; FIG. 2-C shows the results of X-ray, MICRO-CT and MRI analyses of OSA in mice homozygous defective for p53 gene; FIG. 2-D is the results of the effect of IU1 on the number of cancer types in wild-type mice, p53 heterozygous knockout mice and p53 homozygous knockout mice; FIG. 2-E is the results of the number of mice with MLT or OSA in p53 heterozygote knockout mice and p53 homozygote knockout mice; CTRL, control; MLT, thymic malignant lymphoma; MOCK, untreated mouse; NA, not applicable; OSA, osteosarcoma; STS, soft tissue sarcoma; WT, wild type;
FIG. 3 is a graph showing the effect of IU1 on the regulation of p 53-dependent mechanism of USP 14; FIG. 3-A is the protein level of a marker associated with Western blotting used to detect cell cycle, senescence and apoptosis in p53 heterozygous knockout mice; FIGS. 3-B and 3-C are graphs showing the results of flow cytometry analysis of the effect of p53 heterozygote knockout mouse IU1 on cell cycle (FIG. 3-B) and distribution (FIG. 3-C); data shown are mean ± SDS, statistical analysis was performed using one-way anova (P <0.05 and P <0.01 compared to control);
FIG. 4 is a graph of the results of the indirect upregulation of p53 by IU1 through inhibition of USP14
FIG. 5 is a graph of the results of IU1 effect on COPS5 induced downstream effectors in vitro and in vivo; fig. 5-a and 5-B are graphs of the expression of p53, USP14 and COSP5 and their correlation results in primary tumor tissue from p53 homozygous knockout mice treated with dimethylsulfoxide (fig. 5-a, CTRL, N ═ 28) or IU1 (fig. 5-B, N ═ 28); FIG. 5-C is a graph showing the results of Western blotting detecting protein levels of downstream effectors of USP14, COSP5 and COSP5 in p53 homozygous knockout mice; data shown are mean ± SDS. Statistical analysis was performed using one-way anova (P <0.05 and P <0.01 compared to control).
Detailed Description
The invention provides application of IU1 with a structure shown as a formula I in preparation of a drug for treating p53 defective tumors.
In the present invention, the IU1 is a cell permeable, reversible proteosome (proteasome) selective inhibitor that specifically inhibits the deubiquitinating activity of USP 14. The molecular weight of the IU1 is 300.37, and the IU1 is not particularly limited in the invention, and the IU1 is well known in the field. In the present examples, IU1 was purchased from Selleck (Shanghai blue Wood chemical Co., Ltd., China, a subsidiary of the Shanghai Selleck Chemicals, USA).
In the present invention, said IU1 preferably achieves the purpose of treating p53 deficient tumors by specifically inhibiting the deubiquitinating activity of USP 14. The results of the study indicate that the effect of IU1 can rescue p53 protein levels, block p53 ubiquitin degradation by USP14 dependent deubiquitinating of COPS5 and modulating p 53. Given that IU1 has been in clinical evaluation of advanced solid malignancies, preclinical findings may also be applicable to human clinics for patient use.
The following examples are provided to illustrate the application of IU1 of the present invention in the preparation of drugs for treating p53 deficient tumors, but they should not be construed as limiting the scope of the present invention.