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Application of SRC inhibitor and FAK inhibitor in preparation of medicine for inhibiting lung cancer metastasis

A technology of lung cancer metastasis and inhibitor, applied in the field of biomedicine, can solve the problem of unclear molecular mechanism of lung cancer metastasis, and achieve the effect of improving clinical treatment effect and inhibiting migration and invasion.

Active Publication Date: 2022-04-08
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the specific molecular mechanism of lung cancer metastasis is still unclear. Therefore, the development of targeted drugs for metastatic lung cancer is imminent

Method used

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  • Application of SRC inhibitor and FAK inhibitor in preparation of medicine for inhibiting lung cancer metastasis
  • Application of SRC inhibitor and FAK inhibitor in preparation of medicine for inhibiting lung cancer metastasis
  • Application of SRC inhibitor and FAK inhibitor in preparation of medicine for inhibiting lung cancer metastasis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Tumor tissues from NSCLC patients were collected, including primary lung tumors, lymph nodes, brain, and pleural metastases, and after making paraffin sections, immunohistochemical staining was performed to detect the expression of SRC protein and FAK protein. After baking, dewaxing, hydration, and blocking, add SRC or FAK primary antibody working solution dropwise and incubate overnight. The next day, add pika universal secondary antibody working solution dropwise for incubation, and add DAB (diaminobenzidine) for color development. After counterstaining with hematoxylin, hydrochloric acid alcohol differentiation, dehydration and transparency, observe under the microscope. The expression of SRC protein is as follows figure 1 As shown, the expression of FAK protein is as follows figure 2 shown. Depend on figure 1 It can be seen that compared with the primary lung lesions, the levels of SRC protein in lymph node metastases, brain metastases and pleural metastases we...

Embodiment 2

[0040]Using Flag and HA as fusion protein tags, the interaction between SRC protein and FAK protein was studied by co-immunoprecipitation (Co-IP) and western blot (Western blot, WB). Construct the truncated expression plasmids of the full length of SRC or FAK and different structural domains. After transfecting 293T cells, use RIPA buffer and protease inhibitor mixture to extract the protein, then add 20 μL protein A / G agarose beads, and incubate at 25°C 1h, to remove non-specific binders. Collect the supernatant, mix it with the mixture containing Flag antibody and HA antibody (1μg), incubate overnight at 4°C, then add 50μL protein A / G agarose beads, incubate for 2h, and then wash with fresh cold RIPA buffer 3 times to obtain protein precipitation. The protein precipitate was dissolved in the loading buffer and separated by SDS-PAGE. The experimental results of co-immunoprecipitation and western blotting were as follows: Figure 5 shown. Depend on Figure 5 It can be seen...

Embodiment 3

[0042] To investigate the influence of SRC / FAK in downstream pathways that promote tumor metastasis, western blot analysis was performed using NSCLC cells. The human NSCLC cell lines H1975 and H460 used in this example were purchased from the American Type Culture Collection (ATCC) in 2015. The above NSCLC cells were stored in DMEM medium or RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), penicillin (100 μg / mL) and streptomycin (100 μg / mL), and placed in an incubator At 37°C, containing 5% CO 2 , and cultured under saturated humidity conditions, and the cells with a passage number less than 5 were collected for experiments.

[0043] Among them, NSCLC cells without any inhibitor treatment were used as the control group, and NSCLC cells treated with KX2-391 and PF-573228 were used as the experimental group, and the results were as follows: Figure 6 shown. Depend on Figure 6 It can be seen that the downstream signaling pathways of SRC prote...

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Abstract

The invention provides application of an SRC inhibitor and an FAK inhibitor in preparation of drugs for inhibiting lung cancer metastasis. Researches find that the SRC protein and the FAK protein play an extremely important role in lung cancer metastasis, and both the SRC protein and the FAK protein are potential lung cancer prognosis biomarkers and are promising treatment targets, especially under the condition of lymph node or remote metastasis. The SRC inhibitor and the FAK inhibitor are combined for use, so that signal channels (such as an epithelial-mesenchymal transition signal channel and an angiogenesis signal channel) related to lung cancer cell metastasis can be inhibited, and migration and invasion of lung cancer cells are inhibited; the prepared medicine has a remarkable inhibition effect on lung cancer cell metastasis. The method is of great significance in improving the clinical treatment effect of metastatic lung cancer.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of SRC inhibitors and FAK inhibitors in the preparation of drugs for inhibiting lung cancer metastasis. Background technique [0002] Lung cancer is one of the malignant tumors with the highest morbidity and mortality worldwide. According to statistics, in 2020, the global morbidity and mortality of lung cancer will rank second and first respectively. Lung cancer is mainly divided into two pathological types, non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC), among which, NSCLC accounts for about 85% of lung cancers, and SCLC accounts for about 15% of lung cancers. When feasible, surgery remains the mainstay of treatment for lung cancer. Although targeted therapy and immunotherapy have made some clinical progress in recent years, nearly 70% of NSCLC patients already have metastases at the time of diagnosis, which leads to the loss of surgical oppor...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K31/5377A61K31/506A61P11/00A61P35/00A61P35/04
CPCY02A50/30
Inventor 云径平付凌怡
Owner SUN YAT SEN UNIV CANCER CENT
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