An inhibitor targeting ets domain protein and its application
A domain protein and inhibitor technology, applied in the field of inhibitors targeting ETS domain proteins, can solve problems such as undiscovered
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Embodiment 1
[0033] Source of experimental materials:
[0034] Recombinant Human Transcriptional regulator ERG was purchased from CUSABIO Company; SeriesS Sensor Chip CM5 was purchased from Cytiva Company; Nifuratel, Nifuroxazide, Nifursol, Andrographolide, Olaparib and CCK8 were purchased from MCE; transcriptome sequencing was completed by Novogene Company.
[0035] Cell proliferation inhibition assay:
[0036] Wait for 75cm 2 When the cell growth confluence in the culture flask reaches about 85%, the cells are digested, and 5000 VCap cells per well are seeded in a 96-well plate, and the complete culture medium is 100 μl per well. After the cells adhered, different concentrations of Nifuratel, Nifuroxazide, Nifursol, and Andrographolide solutions prepared in complete medium were added, and the same amount of solvent DMSO was used as a control, and incubated in an incubator for 72 hours. Remove the upper medium, add 10 μl of CCK8 stock solution and 90 μl of the corresponding complete med...
Embodiment 2
[0042] The source of the experimental material is the same as that of Example 1, and the following uses Nifuroxazide as the representative of the acetyl hydrazide derivative and its analogs of the present invention, hereinafter referred to as NFZ, to carry out the experimental test. Although no specific experiments have been carried out on other acetylhydrazide derivatives with similar structures and their analogs, those in the art know that familiar experimental results can be obtained.
[0043] 1. NFZ and ERG affinity determination
[0044] According to the regulations of USP40 IMMUNOLOGICAL TEST METHODS—SURFACE PLASMONRESONANCE (1105), ERG protein was immobilized on CM5 chip, and NFZ was prepared into 100, 50, 25, 12.5, 6.25, 3.125, 1.5625 and 0 μM / L through the experimental channel in turn. , with the same amount of solvent DMSO as a reference, and the detection was completed by Biacore8K. The result is as figure 1 shown.
[0045] according to figure 1 , and the KD val...
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