Application of PF-4708671 to preparation of drugs for reversing osimertinib resistance of non-small-cell lung cancer

1. PF-4708671, non-small cell lung cancer technology, applied in the field of genetic engineering, to improve survival rate, good clinical promotion value, and reduce drug resistance

Inactive Publication Date: 2019-12-06
SIR RUN RUN HOSPITAL NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Osimertinib resistance is currently an important bottleneck affecting the treatment outcome and survival time of lung cancer patients. Existing studies can only partially clarify the mechanism of osimertinib resistance, but there are few studies on new methods for reversing drug resistance. Therefore, how to reverse or delay osimertinib resistance can greatly prolong the survival time of lung cancer patients and save patients' treatment costs, which has important clinical significance and social benefits

Method used

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  • Application of PF-4708671 to preparation of drugs for reversing osimertinib resistance of non-small-cell lung cancer
  • Application of PF-4708671 to preparation of drugs for reversing osimertinib resistance of non-small-cell lung cancer
  • Application of PF-4708671 to preparation of drugs for reversing osimertinib resistance of non-small-cell lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The influence of PC-9, PC-9 / G, HCC-827, HCC-827 / OR on Osimertinib sensitivity by CCK-8 method:

[0029] 1) Cell inoculation: Digest monolayer cultured cells with 0.25% trypsin, prepare a single cell suspension with 1640 culture medium containing 10% fetal bovine serum, inoculate 2500 cells per well in a 96-well culture plate, each well Volume 200ul.

[0030] 2) Culture cells: move the culture plate into CO 2 In an incubator at 37°C, 5% CO 2 : and relative humidity conditions, cultivated for 3 days.

[0031] 3) Color development: After the first 1-3 days of culture, add 20ul of CCK-8 solution to each well, and continue to incubate at 37°C for 4h.

[0032] 4) Colorimetry: Measure the optical density (OD) at a wavelength of 490nm, and repeat the experiment three times with 3 replicate wells in each group. Calculate the survival rate. Survival rate = OD value of the experimental group / OD value of the control group × 100%.

[0033] Seed pPC-9, PC-9 / G, HCC-827, HCC-827 / OR...

Embodiment 2

[0037] The activation level of p-s6k1 in PC-9, PC-9 / G, HCC-827, HCC-827 / OR cells was detected by Western blot:

[0038] 1. Method: extraction of total cell protein; SDS-PAGE electrophoresis; semi-dry transfer; western blot; ECL detection

[0039] 2. Results determination: if image 3 shown.

[0040] 3. Results analysis: the expression level of p-s6k in osimertinib-resistant PC-9 / G, HCC-827 / OR cell lines was significantly higher than that in osimertinib-sensitive PC-9, HCC-827 cell lines .

Embodiment 3

[0042] Sensitivity of HCC-827 / OR cells to osimertinib and osimertinib+PF-4708671 detected by CCK-8 method

[0043] 1) Cell inoculation: Digest monolayer cultured cells with 0.25% trypsin, prepare a single cell suspension with 1640 culture medium containing 10% fetal bovine serum, inoculate 2500 cells per well in a 96-well culture plate, each well Volume 200ul.

[0044] 2) Culture cells: move the culture plate into CO 2 In an incubator at 37°C, 5% CO 2 : and relative humidity conditions, cultivated for 3 days.

[0045] 3) Color development: After the first 1-3 days of culture, add 20ul of CCK-8 solution to each well, and continue to incubate at 37°C for 4h.

[0046] 4) Colorimetry: Measure the optical density (OD) at a wavelength of 490nm, and repeat the experiment three times with 3 replicate wells in each group. Calculate the survival rate. Survival rate = OD value of the experimental group / OD value of the control group × 100%.

[0047] HCC-827 / OR treated with osimertin...

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Abstract

The invention belongs to the field of genetic engineering, and particularly relates to application of PF-4708671 to preparation of drugs for reversing drug resistance of non-small-cell lung cancer toosimertinib. By constructing a non-small-cell lung cancer cell strain with the drug resistance to the osimertinib and fluorescent in-situ lung cancer mouse model, it is found that the PF-4708671 can achieve the effect of reversing the drug resistance of lung cancer cells to the osimertinib at the cell and animal levels.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and particularly relates to the application of PF-4708671 in the preparation of drugs for reversing the drug resistance of non-small cell lung cancer to osimertinib. Background technique [0002] Lung cancer is currently the leading cause of death among men and women suffering from cancer in the world, and non-small cell lung cancer (NSCLC) accounts for 85% of all lung cancer patients. At present, the annual number of new cases and deaths of non-small cell lung cancer in the world is constantly rising, and the majority of medical staff have also brought severe tests. Curative surgical resection is usually only suitable for early onset. The five-year survival rate of lung cancer in various stages is only 15%. For patients with limited-stage disease detected, the survival rate is only 49%, which shows that even with early diagnosis, recurrence and death of the disease are very common, therefore...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/506A61P35/00
CPCA61K31/506A61P35/00
Inventor 沈华吴卫兵朱陵君屈晓燕许晶刘锦源
Owner SIR RUN RUN HOSPITAL NANJING MEDICAL UNIV
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