New medical use of cucurbitacin

A new technology of cucurbitacin, applied in the field of medicine, achieves low toxicity and obvious effect

Inactive Publication Date: 2009-06-17
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the anti-leukemia effect and mechanism of cucurbitacin has not been reported at home and abroad.

Method used

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  • New medical use of cucurbitacin
  • New medical use of cucurbitacin
  • New medical use of cucurbitacin

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a new medicament application of cucurbitacins, namely an application of mixing effective monomers of cucurbitacins in medicament for treating leukaemia or an application of effective monomers of cucurbitacins to independently treating leukaemia, belonging to the technical field of medicament. The cucurbitacins with nano-mole concentration can obviously inhibit multiplication of leukaemia HL60 and K562 cells; the cucurbitacins have no cell toxic effect to human leukaemia cells in a multiplication inhibiting concentration range; and the multiplication inhibiting effect to leukaemia cells in vitro is stronger than that to solid tumour cells. Further researches suggest that the cucurbitacins with micro-mole concentration can induce apoptosis of leukaemia HL60 cells by increasing cucurbitacin concentration. Anti-tumour researches in vivo suggest that the cucurbitacins can prolong survival time of a small mouse P388 leukaemia animal model. The effective monomers of cucurbitacins in resisting leukaemia have obvious effect, low toxicity, long lasting drug administration, can be used for both chemotherapy and radiotherapy, and are promising to be further developed into the medicament for treating leukaemia.

Description

Technical field: [0001] The invention belongs to the technical field of medicine, and relates to a new medical application of cucurbitacin, in particular to the application of cucurbitacin in the treatment of human leukemia, that is, the application of cucurbitacin effective monomer components alone or in mixture in the treatment of leukemia. Background technique: [0002] Leukemia is a malignant disease of hematopoietic tissue, commonly known as "blood cancer", which is characterized by the tumorous proliferation of a certain type of leukemia cells in bone marrow or other hematopoietic tissues, which can infiltrate various organs and tissues in the body and affect the function of various organs. damage, resulting in corresponding symptoms and signs. In my country, there are about 3 to 4 leukemia patients per 100,000 population. Leukemia has the highest incidence of malignant tumors in children. According to surveys, the incidence of leukemia in children <10 years old in ...

Claims

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Application Information

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IPC IPC(8): A61K31/575A61P35/02
Inventor 马恩龙李艳春
Owner SHENYANG PHARMA UNIVERSITY
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