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Method for screening anti-breast cancer metastasis compounds, and applications of related compounds

An anti-breast cancer and compound technology, applied in biochemical equipment and methods, microbiological measurement/inspection, medical preparations containing active ingredients, etc., can solve problems such as unsuitability, high cost, and difficulty in customizing microarray chips

Active Publication Date: 2018-03-27
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] With the rapid development of high-throughput biotechnology, both microarray chips and next-generation sequencing technologies can easily detect changes in gene expression profiles after drug treatment, but there are difficulties in customizing microarray chips, and one chip can only measure a small amount. Samples, high cost and other disadvantages are not suitable for high-throughput detection of characteristic gene expression profiles

Method used

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  • Method for screening anti-breast cancer metastasis compounds, and applications of related compounds
  • Method for screening anti-breast cancer metastasis compounds, and applications of related compounds
  • Method for screening anti-breast cancer metastasis compounds, and applications of related compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Discovery of compounds that reverse breast cancer lung metastasis gene profile

[0055] In 2005, the laboratory of Joan Massague in the United States reported that 18 genes were closely related to lung metastasis of breast cancer, of which 15 genes were highly expressed in cells with strong lung metastasis ability of breast cancer, and 3 genes were low expressed, as shown in Table 1. It suggests that if the expression of these 18 genes is reversed in cells with strong lung metastasis ability of breast cancer, it is possible to inhibit the occurrence of lung metastasis of breast cancer.

[0056] Table 1: Names, functional classification and expression of genes related to lung metastasis of breast cancer

[0057]

[0058]

[0059] Through large-scale compound screening, we expect to find active compounds that can reverse the expression direction of the above genes in breast cancer lung metastasis cells. Specific steps are as follows:

[0060] 1. Design ...

Embodiment 2

[0106] Example 2: Compound F (GSK1070916, N'-[4-[4-[2-[3-[(dimethylamino)methyl]phenyl]-1H-pyrrolo[2,3-B]pyridine -4-yl]-1-ethyl-1H-pyrazol-3-yl]phenyl]-N,N-dimethylurea), compound G (Ponatinib, Ponatinib) inhibit MDA-MB-231 cell migration assay

[0107] MDA-MB-231 is a breast cancer cell line commonly used in experiments. It has fast growth, strong cell migration ability, and also has high metastasis ability in animal experiments. Cell migration is an essential step in cancer metastasis. Inhibiting cancer cell migration can effectively inhibit cancer cell metastasis.

[0108] Add 800 μL of 1640 medium containing 10% FBS and 1 μM compound into wells of a 24-well culture plate. Place the Transwell chambers on a 24-well plate. MDA-MB-231 cells cultured to the logarithmic phase were taken, digested with trypsin, and centrifuged to collect the cell pellet. Adjust the cell concentration to 2.5 × 10 with serum-free 1640 medium containing 1 μM compound 5 cells / mL, slowly and eve...

Embodiment 3

[0113] Example 3: Compound A affects the cell cloning ability of MDA-MB-231

[0114] The cloning ability of cells is an important guarantee for the survival and growth of cancer cells after metastasis to distant sites, and is an important indicator for evaluating cancer metastasis.

[0115]The MDA-MB-231 cells cultured to the logarithmic phase were taken, digested with trypsin, and the cell pellet was collected. The cells were suspended in 1640 medium containing 10% FBS and 1% double antibody, and seeded in a 6-well plate with 500 cells per well. After 12 hours, add DMSO to the culture medium of one group, add compound A to the final concentration of 1 μM, 0.33 μM, and 0.11 μM respectively in one group, culture in a CO2 incubator at 37°C, and change the medium containing DMSO or compound A every 3 days . On the 10th day, discard the medium, and stain with 0.1% crystal violet for 30 min. Wash with PBS after staining, take pictures with imager, and each experiment has 3 biolo...

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Abstract

The present invention provides a method for screening anti-breast cancer metastasis compounds by using a tumor metastasis gene map. The method comprises: randomly selecting at least one gene from 18 genes such as MMP1, FSCN1, PTGS2, EREG, ANGPTL4, VCAM1, CXCR4, CXCL1, RARRES3, ID1, TNC, LY6E, LTBP1, C10orf116, KRTHB1, KYNU, MAN1A1 and NEDD9; and designing probes, and selecting compounds for converting the expression levels of the genes in breast cancer cells after metastasis to the expression levels of the genes in in-situ carcinoma cells or normal cells. According to the present invention, the 20 compounds are selected through the method, and the cytological level and animal level experiment results demonstrate that the compounds or the combinations thereof exhibit the anti-breast cancermetastasis activity to different degrees.

Description

technical field [0001] The invention relates to the field of medicines, in particular to a method for screening anti-breast cancer metastasis compounds and the application of related compounds. Background technique [0002] Breast cancer is one of the most common malignant tumors in women. Every year, about 1.3 million people in the world are diagnosed with breast cancer, and about 400,000 people die from the disease. In the past 20 years, the incidence of breast cancer in my country has been increasing year by year, ranking first in the incidence of female malignant tumors, seriously endangering the health of women. Metastasis is one of the hallmarks of tumor malignant biology and the main cause of tumor treatment failure and patient death. [0003] Tumor metastasis is a multi-step, multi-factor involved complex biological process, which includes multiple steps, including: 1. Tumor cells proliferate and fall off from the primary site; 2. Break through blood vessels and ent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11A61K38/12A61K31/23A61K31/4409A61K31/365A61K31/353A61K31/5025A61K31/437A61K31/5377A61K31/7048A61K31/166A61K31/192A61K31/164A61K31/7036A61K31/352A61K31/12A61K31/546A61K31/519A61K31/4545A61P35/00
CPCA61K31/12A61K31/164A61K31/166A61K31/192A61K31/23A61K31/352A61K31/353A61K31/365A61K31/437A61K31/4409A61K31/4545A61K31/5025A61K31/519A61K31/5377A61K31/546A61K31/7036A61K31/7048A61K38/12C12Q1/6886C12Q2600/136C12Q2600/158A61K2300/00C12Q1/68C12N15/11A61P35/00
Inventor 邵伟李莎莎王栋
Owner TSINGHUA UNIV
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