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
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[0035] Example 1:
[0036] The 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, 0.1μg / 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, 0.1μg / mL , The final concentration is 0, 20, 50, 100μM;
[0038] The cells in the logarithmic growth phase are seeded in a 96-well plate at a density of 8000 cells / well. After the cells adhere to the wall, the drug-containing solution is given at the above experimental concentration, 100 μL per well, and three multiple wells for each concentration. Then place the cell plate in a 37°C incubator for 24 hours. After the incubation is over, gently aspirate the culture solution, add 100 μL of 0.5μg / mL MTT solution (prepared in PBS) to each well, and place it in a 37°C incubato...
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[0041] Example 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 formulated to a concentration of 0.01 μg / mL with 1640 medium containing 10% serum , Add an appropriate amount of ambroxol to the 0.01μg / mL paclitaxel solution to make the final concentration 50μM. The 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, they were given blank medium, 0.01μg / mLPTX, 50μM Ax, 0.01μg / mLPTX+50μM Ax at the above experimental concentrations Solution, 2mL per well, with three replicate wells for each concentration. After the administration, place the cell plate in a 37℃ incubator for 24 hours. After the culture is over, collect the upper culture medium, wash with PBS 3 times, collect the washing liquid, add an appropriate amount of 0.25% try...
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[0043] Example 3:
[0044] Taking nude mice bearing A549 subcutaneous tumor as a model, using tumor volume, tumor weight, and survival time as indicators to evaluate whether the combination of PTX and Ax can improve the therapeutic effect of anticancer drugs:
[0045] Culture A549 cells under conventional conditions. When they grow to 80%-90% confluence, digest, centrifuge and resuspend with PBS to prepare 2×10 7 / mL of single cell suspension, inoculated into the right axillary skin of BALB / c nude mice, each with 200μL. Wait until the tumor grows to 50-100mm 3 At time (denoted as 0 days), the tumor-bearing mice were randomly divided into 6 groups, each with 6 mice, respectively, saline group (saline), high-dose ambroxol group (Ax: 100mg / kg), low-dose paclitaxel 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 The dose of ambroxol group (PTX: 3 mg / kg + Ax: 100 mg / kg) w...
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