New medical use of cucurbitacin
A new technology of cucurbitacin, applied in the field of medicine, achieves low toxicity and obvious effect
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Embodiment 1
[0027] Example 1 In vitro experiments on the inhibitory effect of cucurbitacin E and B on the proliferation of human solid tumor and leukemia cells
[0028] 1 Experimental method:
[0029] HeLa (human cervical cancer cells), BEL7402 (human liver cancer cells), SGC7901 (human gastric cancer cells) and HT1080 (human fibroblastoma cells) were used at 1×10 per well. 4 Inoculate at a density of 24-well plates in RPMI 1640 medium containing 10% fetal bovine serum and 2% glutamine at 37°C, 5% CO 2 After culturing, 24 hours later, different concentrations of cucurbitacin E and B were added to different plates, and a negative control group was set up (adding the same amount of culture solution containing or not containing DMSO). Cells were counted by Trypan blue staining after digestion 72h after drug addition.
[0030] Leukemia cells K562 and HL60 were used at 2×10 per well 4 Each / ml density is inoculated in 24 well plate, then add the medicinal liquid of different concentration re...
Embodiment 2
[0033] Example 2 Effects of cucurbitacin B and E on the survival period of mouse P388 leukemia model animals in vivo
[0034] 1 Experimental method:
[0035] DAB / 2 mice were divided into 5 groups, namely the control group (administered with 0.5% CMC-Na), the low-dose cucurbitacin E group, the high-dose cucurbitacin E group, the low-dose cucurbitacin B group, and the high-dose cucurbitacin B group . Animals in each group were inoculated with 2×10 P388 leukemia cells in the peritoneal cavity. 6 Each animal, 24 hours after inoculation, intraperitoneally inject the corresponding dose of medicine every day, the volume of administration is 0.1ml / kg, and the administration is continuous for 7 days. After completion, the animals are raised normally, and the survival time of animals in each group is recorded and the life is calculated. elongation rate. Life extension rate (%)=[(average survival time of animals in administration group-average survival time of animals in control group...
Embodiment 3
[0040] Example 3 Flow cytometry detection of cucurbitacin E on human leukemia K562, HL60 cell cycle and cycle regulatory proteins
[0041] 1 Experimental method:
[0042] 1.1 The effect of cucurbitacin E on the cell cycle of human leukemia K562 and HL60 detected by flow cytometry
[0043] HL60 and K562 cells at 1×10 6 Cells were planted in a culture flask at a density of 1 / ml. After 12 hours of adding the drug, the cells were collected and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded carefully, and 500 μl of PBS was added to suspend the cells, and 10 ml of 70% ice-cold ethanol was added dropwise. Fix overnight at -20°C. Centrifuge the sample before detection at 2000 rpm / min for 5 minutes, carefully discard the supernatant, add 10 μl of RNAase (200 μg / ml) to each sample, mix well, and act at 37°C for 30 minutes, then add 50 μl of PI (1 mg / ml) , 4°C in the dark for 30 minutes, flow cytometry detection, counting 10,000 cells per sample.
[0044] 1.2 We...
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