Application of echinacoside in anti-tumor medicaments
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An anti-tumor drug, echinacoside technology, applied in the direction of anti-tumor drugs, drug combinations, medical preparations containing active ingredients, etc.
Active Publication Date: 2015-01-21
JILIN UNIV
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Problems solved by technology
However, in foreign countries, the development of Chinese medicine is hindered abroad due to the unknown active ingredients and mechanism of action in traditional Chinese medicine.
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Embodiment 1
[0010] We added MTH1, inorganic pyrophosphatase and dGTP into the reaction solution (100mM pH8.0 Tris-acetic acid, 40mM NaCI, 10mM magnesium acetate, 0.005% Tween20 and 1mMDTT). The final concentration of each component is 1nM MTH1, 100μM dGTP and 0.2U ml-1 inorganic pyrophosphatase, the concentration of echinacoside is 3μM, 6μM, 9μM, 12μM, 15μM, and the experiment is carried out in a 96-well plate with a volume of 100μl. The enzyme and substrate were incubated at room temperature for 1 hour, and 25 μl malachite green solution was added to terminate the reaction, and the absorbance was measured at 630 nm.
[0011] from figure 1 It can be seen in the enzymatic experiments that echinacoside has a better inhibitory effect on MTH1, and the inhibitory effect is dose-dependent with the increase of drug concentration. The half maximal inhibitory concentration (IC) of echinacoside on MTH1 50 ) was 7.42±2.13 μmol / L.
Embodiment 2
[0013] In order to detect the effect of echinacoside on tumor cells at the cellular level, we selected SW480 human colon cancer cells and U2OS human osteosarcoma cells for experiments. First, dilute echinacoside into different concentrations according to the concentration gradient, spread appropriate amount of cells on a 96-well plate, add echinacoside when the growth density reaches about 80%, and the final concentrations are 15 μM, 25 μM, 35 μM, 45 μM, 55 μM , 65 μM, 75 μM, 85 μM, 95 μM, the reaction volume is 100 μl, after 24 hours of drug action, add MTT to react for 4 hours, add 150 μl DMSO, and detect the absorbance value at 630nm with a microplate reader. The above experiments were carried out at 37°C, 5% CO 2 cultivated under conditions.
[0014] The result is as figure 2 with image 3 As shown, after adding echinacoside, the cell viability decreased with the increase of echinacoside concentration. It shows that echinacoside can inhibit the growth of SW480 and U2O...
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Abstract
The invention relates to application of echinacoside in anti-tumor medicaments. The application provided by the invention is as follows: firstly adding MTH1, inorganic pyrophosphatase and dGTP into a reaction solution (100mM pH 8.0 Tris-acetic acid, 40mM NaCl, 10mM magnesium acetate, 0.005% Tween 20 and 1mM DTT), incubating an enzyme and a substrate at room temperature for 1h, then adding 25 mu l of malachite green solution to terminate reaction and performing absorbance detection at 630nm by using an iMark microplate reader, wherein results show that echinacoside has a significant effect of inhibiting enzyme activity of MTH1; and secondly detecting the effects of echinacoside against tumor cells at the cellular level by an MTT experiment, wherein the results show that echinacoside can obviously inhibit the growth of SW480 colon cancer cells and U2OS human osteosarcoma cells. According to the application provided by the invention, echinacoside is used for inhibiting specific enzyme MTH1 for maintaining survival in the tumor cells, and oxidized nucleotide is mixed into DNA, thereby resulting in fatal DNA double-strand break in the cancer cells, producing an anti-tumor effect, finding a new medical use of echinacoside and laying a foundation for future development of the new anti-tumor medicaments.
Description
technical field [0001] The invention belongs to the field of application of traditional Chinese medicines, and in particular relates to the application of echinacoside in the preparation of antitumor drugs. Background technique [0002] Cancer is the leading cause of death in animals and humans, and it seriously threatens human life and health. People have been making unremitting efforts to obtain active and safe antineoplastic drugs for cancer patients. We found for the first time that echinacoside can inhibit the specific enzyme MTH1 in tumor cells to maintain survival, leading to the incorporation of oxidized nucleotides into DNA, resulting in lethal DNA double-strand breaks in cancer cells, in order to produce anti-tumor cells. tumor effect. [0003] Traditional Chinese medicine has anti-virus history of thousands of years, and my country has accumulated a lot of valuable theoretical and practical experience in the use of traditional Chinese medicine to prevent and tre...
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