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Application of ambroxol to preparation of tumor chemotherapy drug synergistic agent

A chemotherapeutic drug, the technology of ambroxol, applied in the field of biomedicine, can solve the problems such as the inability of cells to maintain the basic structure, reduce the sensitivity of tumor cells, reduce the effect of chemotherapy treatment, etc., achieve inhibition of tumor growth, good clinical transformation prospects, and good resistance The effect of tumor effect

Inactive Publication Date: 2019-04-05
FUDAN UNIV
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Problems solved by technology

On the one hand, the occurrence of autophagy in tumor cells can serve as a survival mechanism to protect tumor cells and inhibit apoptosis; on the other hand, the continuous activation of autophagy makes cells unable to maintain their basic structure, leading to tumor cell death, that is, autophagic death.
For example, fluorouracil acting on human colon cancer cells can induce protective autophagy of tumor cells, reduce the sensitivity of tumor cells to drugs, and even drug resistance; while paclitaxel (PTX) induces tumor cell apoptosis, the The concentration of the drug can weaken the phosphorylation of mTOR and its downstream substrate p70s6k, the autophagy-related protein of tumor cells, and both Beclin1 and Atg5 are up-regulated, suggesting that paclitaxel initiates the production of tumor cell autophagy through Beclin1, leading to drug resistance and reducing chemotherapy. treatment effect

Method used

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  • Application of ambroxol to preparation of tumor chemotherapy drug synergistic agent
  • Application of ambroxol to preparation of tumor chemotherapy drug synergistic agent
  • Application of ambroxol to preparation of tumor chemotherapy drug synergistic agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] MTT method was used to evaluate the toxicity of different concentrations of paclitaxel combined with ambroxol incubating A549 cells:

[0037] Before the experiment, paclitaxel was prepared into a series of concentrations of 0.001, 0.01, 0.05, and 0.1 μg / mL with 1640 medium containing 10% serum, and an appropriate amount of ambroxol was added to the paclitaxel solution of 0.001, 0.01, 0.05, and 0.1 μg / mL , so that the final concentrations are 0, 20, 50, and 100 μM, respectively;

[0038] The cells in the logarithmic growth phase were seeded in a 96-well plate at a density of 8000 cells / well. After the cells adhered to the wall, the drug-containing solution was given according to the above experimental concentration, 100 μL per well, and three replicate wells were set up for each concentration. Afterwards, place the cell plate in a 37°C incubator for 24 hours. After the end of the culture, gently aspirate the culture medium, add 100 μL of 0.5 μg / mL MTT solution (prepared ...

Embodiment 2

[0042] The Annexin V-FITC / PI apoptosis detection kit was used to evaluate the apoptosis level of A549 cells incubated with different concentrations of paclitaxel combined with ambroxol: before the experiment, paclitaxel was prepared at a concentration of 0.01 μg / mL in 1640 medium containing 10% serum , Add an appropriate amount of ambroxol to the 0.01 μg / mL paclitaxel solution to make the final concentration 50 μM. Cells in the logarithmic growth phase were seeded in a 6-well plate at a density of 50,000 cells / well. After the cells adhered to the wall, the blank medium, 0.01 μg / mLPTX, 50 μM Ax, 0.01 μg / mLPTX+50 μM Ax were given according to the above experimental concentrations Solution, 2mL per well, set up three duplicate wells for each concentration. After administration, the cell plate was cultured in a 37°C incubator for 24 hours. After the culture was over, the supernatant culture fluid was collected, washed with PBS for 3 times, and the washing fluid was collected, and ...

Embodiment 3

[0044] Using nude mice bearing A549 subcutaneous tumors as models, tumor volume, tumor weight, and survival time were used as indicators to evaluate whether the combination of PTX and Ax could improve the therapeutic effect of anticancer drugs:

[0045] A549 cells were cultured according to conventional conditions, and when they grew to 80%-90% confluence, they were digested and centrifuged and resuspended with PBS to prepare 2×10 7 / mL single-cell suspension, inoculated into the subcutaneous right axilla of BALB / c nude mice, 200 μL each. When the tumor grows to 50-100mm 3 At day 0 (denoted as day 0), the tumor-bearing mice were randomly divided into 6 groups, 6 mice in each group, including normal saline group (saline), high-dose ambroxol group (Ax: 100mg / kg), low-dose paclitaxel group group (PTX:3mg / kg), low-dose paclitaxel+low-dose ambroxol group (PTX:3mg / kg+Ax:20mg / kg), high-dose paclitaxel group (PTX:10mg / kg), low-dose paclitaxel+high Dose Ambroxol group (PTX: 3mg / kg+Ax...

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Abstract

The invention belongs to the technical field of biological medicine, and relates to new application of ambroxol to pharmacy, in particular to application of the ambroxol to preparation of a tumor chemotherapy drug synergistic agent. In application, the ambroxol can enhance the lethal effect of various chemotherapy drugs such as paclitaxel, docetaxel, adriacin, cis-platinum and camptothecin on various tumor cells such as A549, MCF-7 and B16F10; an A549 subcutaneous tumor model and an A549-luciferase lung metastasis tumor model of a nude mouse are established, an ambroxol and the paclitaxel combined treatment scheme is used, enlarging of a tumor nidus is inhibited more effectively, the survival time of the nude mice is prolonged, and the effect is better if the dose of the ambroxol is higher; and the ambroxol can adjust autophagy of the tumor cells, and enhancing of the lethal effect of the paclitaxel by the ambroxol is possibly related to regulation of autophagy of the cells by the ambroxol. The ambroxol serves as a common drug in clinical treatment of acute bronchitis, chronic bronchitis and bronchial asthma, the toxic and side effects are small, the obvious curative effect is achieved, and combined use of the ambroxol and the paclitaxel for treatment of clinical transformation of tumors, especially lung tumors has good prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to the new use of ambroxol in pharmacy, in particular to the use of ambroxol in the preparation of synergistic preparations of tumor chemotherapy drugs Background technique [0002] According to statistics, with the aging population and environmental pollution, the morbidity and mortality of malignant tumors have been on the rise, seriously threatening human life and health. According to the data of the "Third National Population Cause of Death Survey" released by the health department, the mortality rate of malignant tumors among urban and rural residents in my country is at a high level in the world, and it is showing a continuous growth trend. The traditional methods of treating tumors in clinical practice include surgical resection, chemotherapy, radiotherapy, etc., each of which has certain therapeutic advantages, but also has its limitations, for example: surgical resection h...

Claims

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Application Information

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IPC IPC(8): A61K31/137A61K31/337A61P35/00
CPCA61K31/137A61K31/337A61K2300/00
Inventor 沙先谊张秀磊
Owner FUDAN UNIV
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