Application of apatinib and combination of apatinib and CCI-779 in preparation of drugs for treating lung cancer

A technology of CCI-779 and apatinib, which is applied in the application field of apatinib and CCI-779 in the preparation of lung cancer drugs, which can solve the problems that the role needs to be explored, so as to improve sensitivity and induce apoptosis death effect

Pending Publication Date: 2020-05-08
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Apatinib has been proven to play a role in various types of tumors, such as gastric cancer, liver cancer, breast cancer, colorectal cancer and non-small cell lung cancer, but the effect of apatinib on small cell lung cancer cells needs to be explored

Method used

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  • Application of apatinib and combination of apatinib and CCI-779 in preparation of drugs for treating lung cancer
  • Application of apatinib and combination of apatinib and CCI-779 in preparation of drugs for treating lung cancer
  • Application of apatinib and combination of apatinib and CCI-779 in preparation of drugs for treating lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Cell culture method:

[0030] Take the cells out of the -80°C refrigerator, rewarm quickly in a 37°C water bath, centrifuge at 1000r / min for 3min, discard the supernatant, resuspend the cell pellet with fresh medium, and culture in a medium containing 10% fetal bovine serum. RPMI Medium 1640 medium, placed at 37°C, 5% CO 2 in a humidified incubator. When the cells grow to about 90% confluence, the culture medium is discarded, washed 1-2 times with PBS, digested and passaged with 0.25% trypsin-EDTA. All experiments used cells in logarithmic growth phase.

[0031] CCK8 assay for cell viability:

[0032] Collect NCI-H446 cells that grow stably in the logarithmic phase, count on a cell counting plate, and adjust the concentration of the cell suspension to 2.5×10 4 / mL, add 200 μL of suspension to each well of a 96-well plate, 37°C, 5% CO 2 Cultivate overnight, and after the cells adhere to the wall, set the concentration gradient of Apatinib to 0 μmol / L, 1 μmol / L, 2 μmo...

Embodiment 2

[0036] Cell cycle experiments.

[0037] NCI-H446 cells were inoculated into 6-well plates, and complete medium was added. When the cell confluency reached 70%-80%, the medium in the wells was sucked off, and each well was divided into control group, CCI-779 treatment group, Low concentration of Apatinib treatment group (12μmol / L, 16μmol / L), high concentration of Apatinib treatment group (28μmol / L, 32μmol / L), low concentration of Apatinib (12μmol / L, 16μmol / L) combined with CCI-779 treatment group, high Concentration of Apatinib (28 μmol / L, 32 μmol / L) combined with CCI-779 treatment group, each group was added with different concentrations of drugs 2mL, 37 ° C, 5% CO 2 Incubate for 24h. Collect 1×10 6 Put the cells in the flow tube, wash the cells with pre-cooled PBS, centrifuge at 1000r / min for 3min, discard the supernatant, add 1-2ml of pre-cooled 75% ethanol drop by drop, vortex to mix the cells, and stay overnight at 4°C in the dark , Centrifuge the cells at 2000r / min for...

Embodiment 3

[0040] Apoptosis assay

[0041] NCI-H446 cells were inoculated into 6-well plates, and complete medium was added. When the cell confluency reached 70%-80%, the medium in the wells was sucked off, and each well was divided into control group, CCI-779 treatment group, Low concentration of Apatinib treatment group (12μmol / L, 16μmol / L), high concentration of Apatinib treatment group (28μmol / L, 32μmol / L), low concentration of Apatinib (12μmol / L, 16μmol / L) combined with CCI-779 treatment group, high Concentration of Apatinib (28 μmol / L, 32 μmol / L) combined with CCI-779 treatment group, each group was added with different concentrations of drugs 2mL, 37 ° C, 5% CO 2 Incubate for 24h. Collect 5×10 5 Put the cells in the flow tube, wash the cells with pre-cooled PBS, centrifuge at 1000r / min for 3min, discard the supernatant, add 100μl 1×Binding Buffer to resuspend the cells, add 5μl Annexin V-FITC, and incubate in a 37°C incubator for 10min; 5 μl of PI Staining Solution, mixed gentl...

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Abstract

The invention discloses application of apatinib and a combination of the apatinib and CCI-779 in preparation of drugs for preventing and treating small cell lung cancer, and relates to the technical field of pharmacy. In-vitro tests prove that the apatinib has a concentration-dependent characteristic on the action of a small cell lung cancer cell strain (NCI-H446), and high-concentration apatinibcan inhibit cell proliferation and migration of the small cell lung cancer cell strain and induce cell apoptosis, and can improve the sensitivity of small cell lung cancer cells to the apatinib when combined with an mTOR inhibitor CCI-779, and can inhibit proliferation and migration of the small cell lung cancer cells at a relatively low concentration to cause cell cycle arrest and induce cell apoptosis.

Description

technical field [0001] The invention relates to the technical field of pharmacy, in particular to the application of apatinib and CCI-779 in the preparation of lung cancer medicine. Background technique [0002] Lung cancer is the most common cancer and the leading cause of cancer-related death, and can be divided into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). Among them, SCLC accounts for about 10%-15% of all lung cancers. It is a type of lung cancer with high invasion, high lethality, and widespread metastasis. Hematogenous metastasis can occur in the early stage, and about 60% of patients have distant metastasis when they are diagnosed. 5 The annual survival rate is only 7%, and it is characterized by rapid growth and high vascularization. Angiogenesis is one of the most critical processes in the progression of malignant tumors. It provides nutrients for tumor growth, promotes tumor growth, and is closely related to tumor occurrence, prolifera...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/444A61K31/436A61P35/00
CPCA61K31/444A61K31/436A61P35/00A61K2300/00
Inventor 陈军刘红雨李永文刘超张洪兵张子禾施睿峰徐松林朱光胜
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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