Application of a compound in cancer treatment medicine
A cancer treatment and compound technology, applied in the field of biomedicine, can solve the problems of high heterogeneity drugs in breast cancer that cannot meet targeted and precise treatment
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Embodiment 1
[0043] Example 1: Study of the effect of drugs on cell activity
[0044] The human breast cancer cells MCF-7, ZR-75-1 and T47D cells in the logarithmic growth phase were taken to prepare single cell suspensions, which were inoculated in 96-well culture plates, 180 μl per well, and 12.5, 25, 50, and 100 cells were added to each well. 200 μmol / l of candidate compounds were cultured for 24 h, 48 h, and 72 h, respectively, and 20 μl of MTT was added to each well. After culturing for 4 h, the medium was discarded, and 150 μl of DMSO was added to mix well. The OD value of the well at 570nm wavelength was used to calculate the cell growth inhibition ratio (IR) and IC50.
[0045] In this experimental example, MTT experiments were performed on 14 compounds. In fact, when the inventor was conducting research, the inventor screened 2 million compounds in the library, and screened out a total of 14 potential compounds. In the follow-up MTT experiment, it was further found that four compo...
Embodiment 2
[0055] Example 2: In vitro cytotoxicity evaluation of Zinc-09
[0056] Using the commonly used anticancer drug paclitaxel (PTX) as a positive control, the concentration of PTX was 0.5 μM, and taking normal breast epithelial cells MCF-10A as the research object, the cytotoxicity of Zinc-09 was detected by MTT method. The experimental results are as follows Figure 8 As shown (the data in the figure is in the form of mean±SE, n=6), the results show that in normal mammary epithelial cells MCF-10A, the inhibitory effect of PTX on the cells for 48h is 32.3%, and 200 μM Zinc-09 on MCF-10A The inhibition rate of cells was 40.7%, and when Zinc-09 was lower than 200 μM, the inhibitory effect of the drug on MCF-10A cells was less than that of PTX, indicating that compared with the clinical anti-breast cancer drug PTX, Zinc-09 has less cytotoxicity.
Embodiment 3
[0057] Example 3: Confirmation of the target of Zinc-09
[0058] In this example, conventional western blot was used for detection. The basic process was as follows: cells were inoculated into T25 culture flasks, and after the cells adhered to the wall, different concentrations of test drugs were added to treat them for different times, and then the cells were collected, and RIPA (plus PMSF) was used to collect the cells. After lysing on ice for 1 h, centrifuge at 4 °C for 20 min, take the supernatant, add 5× loading buffer, and cook at 100 °C for 5 min to denature the protein. After separation by SDS-PAGE, the membrane was transferred to PVDF membrane, blocked and incubated with primary and secondary antibodies for color development.
[0059] In the occurrence and development of breast cancer, the expression of SGK3 is regulated by estrogen. Therefore, we first detected 6 breast cancer cell lines (MCF-7, ZR-75-1, T47D, MDA-MB-231, MDA-MB -453, CAL-148) expression of SGK3 pro...
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