Use of yg1702 in the preparation of a drug for resisting ewing's sarcoma

Abstract

The application discloses application of YG1702 in preparation of a drug for resisting Ewing's sarcoma, and YG1702 can effectively inhibit Ewing's sarcoma proliferation, self-renewal and in-vivo tumorigenicity by inhibiting expression of an Ewing's sarcoma ALDH18A1 gene, and can inhibit expression of an EWS-FLI1 fusion gene of the Ewing's sarcoma and downstream target genes EZH2, ID2, PTPL1, CCND1 and VEGFA, and can become a potential target therapy candidate small molecule, and the application has potential clinical value for finding a new Ewing's sarcoma treatment target and improving a combined treatment effect of Ewing's sarcoma patients.

Description

Technical Field

[0001] This invention relates to biomedical technology, specifically to the application of YG1702 in the preparation of drugs for treating Ewing's sarcoma. Background Technology

[0002] Ewing's sarcoma is a malignant bone tumor that commonly affects children and adolescents, with a slight male-to-female prevalence. The standard treatment regimen is high-intensity chemotherapy combined with surgery / radiotherapy. Currently, after receiving cytotoxic therapy, the 5-year survival rate for patients with standard-risk locally acquired Ewing's sarcoma can reach 70-80%, but the 5-year overall survival rate for patients with primary disseminated Ewing's sarcoma remains around 13-30%. More regrettably, approximately 30-40% of patients with locally acquired primary Ewing's sarcoma and 60-80% of patients with primary disseminated Ewing's sarcoma will relapse, and the 5-year survival rate for those who relapse within 24 months of initial diagnosis is less than 10%.

[0003] EWS-FLI1, as an oncogenic factor in Ewing sarcoma, drives extensive epigenetic remodeling and abnormal transcription, but as a disordered protein, it is difficult to target. Therefore, in-depth exploration of the epigenetic and transcriptional regulatory mechanisms of Ewing sarcoma and the search for new therapeutic targets are of significant clinical importance. In recent years, the tumor metabolism-epigenetic axis has attracted much attention due to its potential to provide new strategies for targeted therapy in various malignant tumors. Proline metabolism plays an important role in the tumor metabolism-epigenetic axis due to its involvement in numerous prolyl hydroxylation-mediated epigenetic modifications. ALDH18A1, as a key enzyme in proline metabolism, plays an important "parametrogenic" role in the P5C-proline cycle formed by its catalytic product P5C with proline. High expression of ALDH18A1 is closely related to the self-renewal capacity of Ewing sarcoma.

[0004] Therefore, it is of great significance to explore whether the small molecule inhibitor YG1702 of ALDH18A1 can be used to treat Ewing sarcoma. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide the application of YG1702 in the preparation of a drug for treating Ewing's sarcoma.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] 1. Application of YG1702 in the preparation of drugs for Ewing's sarcoma.

[0008] The preferred application of YG1702 in the preparation of drugs that inhibit the expression of the ALDH18A1 gene in Ewing's sarcoma is described in this invention.

[0009] The preferred application of YG1702 in the preparation of a drug that inhibits the cell proliferation of Ewing's sarcoma cells is described in this invention.

[0010] The preferred application of YG1702 in the preparation of a drug that inhibits the ability of Ewing sarcoma cells to form plate colonies is described in this invention.

[0011] The preferred application of YG1702 in the preparation of drugs that inhibit the spheroidization ability of Ewing sarcoma cells is described in this invention.

[0012] The present invention preferably relates to the use of YG1702 in the preparation of drugs that inhibit the expression of the EWS-FLI1 fusion gene and its downstream target genes in Ewing sarcoma.

[0013] Preferably, the downstream target genes of this invention are EZH2, ID2, PTPL1, CCND1, and VEGFA.

[0014] The preferred application of YG1702 in the preparation of drugs that inhibit the tumorigenicity of Ewing sarcoma cells is described in this invention.

[0015] The beneficial effects of this invention are as follows: This invention discloses the application of YG1702 in the preparation of drugs against Ewing's sarcoma. YG1702 can effectively inhibit the proliferation, self-renewal and tumorigenic capacity of Ewing's sarcoma by inhibiting the expression of the ALDH18A1 gene in Ewing's sarcoma. It can also inhibit the expression of the EWS-FLI1 fusion gene and its downstream target genes EZH2, ID2, PTPL1, CCND1 and VEGFA in Ewing's sarcoma, and may become a potential candidate small molecule for targeted therapy. This invention has potential clinical value for finding new therapeutic targets for Ewing's sarcoma and improving the combined treatment effect for Ewing's sarcoma patients. Attached Figure Description

[0016] To make the objectives, technical solutions, and beneficial effects of this invention clearer, the following figures are provided for illustration:

[0017] Figure 1 In vitro drug sensitivity of YG1702 in Ewing sarcoma cells ( Figure 1 (A) and the effect of YG1702 on the proliferative capacity of Ewing sarcoma cells ( Figure 1 ,B);

[0018] Figure 2 The effect of YG1702 on the plate colony-forming ability of Ewing sarcoma cells ( Figure 2 ,AB);

[0019] Figure 3 The effect of YG1702 on the spheroidization ability of Ewing sarcoma cells ( Figure 3 ,AB);

[0020] Figure 4 The effect of YG1702 on the expression of downstream target genes of EWS-FLI1 in Ewing sarcoma cells. Figure 4,AB);

[0021] Figure 5 The toxic effects of different doses of YG1702 on Ewing sarcoma tissues (heart, liver, kidney, lung, and spleen);

[0022] Figure 6 The effect of YG1702 on the in vivo tumorigenicity of Ewing sarcoma ( Figure 6 AC). Detailed Implementation

[0023] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0024] The structure of YG1702 in this invention is the same as that of YG1702 in the invention patent with application number 201810529687.6.

[0025] Example 1: YG1702's ability to inhibit the cell proliferation of Ewing sarcoma cells

[0026] The IC50 values ​​of YG1702 in A673 and SK-N-MC cells were detected using the CCK8 cell viability assay, and were 0.10 μM ± 0.01 μM and 0.06 μM ± 0.03 μM, respectively. Figure 1 (A). Further growth curves were plotted at the IC50 drug concentration. The results showed that, compared to the control group, the absorbance value of Ewing sarcoma cells at the IC50 concentration of YG1702 was significantly reduced. Figure 1 (B). The above results indicate that the ALDH18A1 inhibitor YG1702 can significantly reduce the in vitro proliferation ability of Ewing sarcoma cells.

[0027] Example 2: YG1702 reduces the plate colony formation ability of Ewing sarcoma cells.

[0028] The colony formation assay was used to evaluate whether the ALDH18A1 inhibitor YG1702 could inhibit the colony formation ability of Ewing sarcoma cells. The results showed that treatment of A673 and SK-N-MC cells with YG1702 significantly reduced the number of colonies formed in Ewing sarcoma cells compared to the control group. Figure 2 The above results indicate that the ALDH18A1 inhibitor YG1702 can significantly inhibit the plate colony formation ability of Ewing sarcoma cells.

[0029] Example 3: YG1702 reduces the spheroidization ability of Ewing sarcoma cells

[0030] Spheroidization experiments showed that after treatment of A673 and SK-N-MC cells with YG1702, the number of cell spheroids formed by Ewing sarcoma cells was significantly reduced compared with the control group. Figure 3 The above results further demonstrate that the ALDH18A1 inhibitor YG1702 can significantly reduce the self-renewal capacity of Ewing sarcoma cells.

[0031] Example 4: YG1702 downregulates the expression of EWS-FLI1 downstream target genes such as PARP1.

[0032] The ALDH18A1 inhibitor YG1702 suppressed the in vitro proliferation, colony formation, and spheroidization abilities of Ewing sarcoma cells. Next, the effects of YG1702 on Ewing sarcoma cells' downstream target genes of EWS-FLI1, such as PARP1, were further investigated. The results showed that YG1702 treatment of A673 cells not only inhibited PARP1 mRNA expression but also extensively downregulated the mRNA levels of multiple EWS-FLI1 downstream target genes, including EZH2, ID2, PTPL1, CCND1, and VEGFA. Figure 4 (Table 1). Similarly, YG1702 treatment of SK-N-MC cells, in addition to inhibiting PARP1 mRNA expression, also extensively downregulated the mRNA levels of EWS-FLI1 downstream target genes, including EZH2, ID2, PTPL1, CCND1, VEGFA, and UPP1. Figure 4 (B)(Table 1). The above results indicate that the ALDH18A1 inhibitor YG1702 can downregulate the expression of downstream target genes of EWS-FLI1, such as PARP1.

[0033] Table 1. Primer sequence information for EWS-FLI1 and its downstream target genes.

[0034]

[0035] Example 5: YG1702 downregulates the expression of EWS-FLI1 downstream target genes such as PARP1.

[0036] Finally, the effect of the ALDH18A1 inhibitor YG1702 on the in vivo tumorigenicity of Ewing sarcoma cells was verified. First, the effects of different doses of YG1702 on mouse organs were examined. HE staining results showed that no significant organ damage was observed at four dosage concentrations: 6.25 mg / kg, 12.50 mg / kg, 25 mg / kg, and 50 mg / kg. Figure 5Furthermore, a nude mouse subcutaneous xenograft model was constructed using the A673 cell line to evaluate the therapeutic significance of YG1702 for Ewing sarcoma. Doxorubicin (Dox) was used as a positive control. Mice were divided into four groups: Control group (solvent group), Dox group, YG1702 group, and combination therapy group (YG+Dox). The Control group received intraperitoneal administration every 2 days, the Dox group received intraperitoneal administration every 5 days (dose: 1.50 mg / kg), the YG1702 group received intraperitoneal administration every 2 days (dose: 50 mg / kg), and the combination therapy group received intraperitoneal administration of YG1702 every 2 days (dose: 25 mg / kg) plus intraperitoneal administration of Dox every 5 days (dose: 0.75 mg / kg). As can be seen, compared with the control group, the tumor volume in the YG1702 group and the combined drug administration group was significantly reduced (the average tumor volume in the control group was 883.90±209.80 mm). 3 The average tumor volume in the YG1702 group was 387.10 ± 72.90 mm. 3 The average tumor volume in the combined drug administration group was 442.30 ± 120.40 mm. 3 Control group vs. YG1702 group, n=6, P=2.70×10 -4 Control group vs. combined drug group, n=6, P=1.20×10 -3 The tumor volume in the Dox group also decreased (the average tumor volume in the Dox group was 586.80 ± 108.60 mm). 3 Control group vs. Dox group, n=6, P=9.41×10 -2 ()( Figure 6 (A, C). Compared with the Control group, the tumor weight in the Dox group, YG1702 group, and combined drug administration group was significantly reduced (mean tumor weight in the Control group was 0.99±0.27g, mean tumor volume in the Dox group was 0.64±0.13g, mean tumor weight in the YG1702 group was 0.28±0.04g, and mean tumor weight in the combined drug administration group was 0.39±0.08g, n=6, P=2.03×10⁻⁶). -3 ()( Figure 6 (B). The above results indicate that YG1702 can effectively inhibit the tumorigenicity of Ewing sarcoma cells and the growth rate of xenografts, and may serve as a candidate small molecule for targeted therapy of Ewing sarcoma.

[0037] The above-described embodiments are merely preferred embodiments provided to fully illustrate the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. Application of YG1702 in the preparation of drugs for Ewing's sarcoma.

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