A kind of alkaloid compound derived from Trichoderma auranthoderma and application thereof
A technology of alkaloids and Trichoderma citrinum, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as drugs that have not yet been seen, and achieve the effect of significant anti-tumor activity
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Embodiment 1
[0014] Fermentative production and separation and purification of the compound of embodiment 1
[0015] 1 Fermentation production
[0016] Fermentation culture of production bacteria: according to the conventional method of cultivating microorganisms, take Trichodermacitrinoviride. IBPT-4 (preserved in China Type Culture Collection Center on January 25, 2013, address: Wuhan University, Wuhan, China, The deposit number is: CCTCCNO: M2013055) appropriate amount, inoculated on the PDA solid slant medium and cultivated in an incubator at 28° C. for 4 days.
[0017] Take an appropriate amount of Trichodermacitrinoviride. IBPT-4 that has been cultured on a slant for 4 days, and inoculate it into 400 mL of culture fluid [Culture fluid composition (g / L): mannitol 20.0, yeast extract 3.0, maltose 20.0, monosodium glutamate 10.0, Glucose 10.0, KH 2 PO 4 0.5, MgSO 4 0.3, NaCl6.0 constant volume] in a 1000mL Erlenmeyer flask, after static culture at 28°C for 30 days, mycelium and ferm...
Embodiment 2
[0027] The test of embodiment 2 in vitro antitumor activity
[0028] 1 Experimental samples and experimental methods
[0029] Preparation of the tested sample solution The test sample is the pure product of the compound separated and purified in the above-mentioned implementation 1. Accurately weigh an appropriate amount of sample, and prepare a solution with the required concentration with methanol for activity measurement.
[0030] Cell lines and cell subculture The human melanoma A375 cell line was used, and the A375 cells were subcultured in RPMI-1640 medium containing 10% FBS at 37°C in an incubator filled with 5% carbon dioxide.
[0031] Cell Proliferation Inhibitory Activity Test Method
[0032] Human melanoma A375 cells in the logarithmic growth phase were obtained by tetrazolium salt (MTT) method, and the cell density was adjusted to 2×10 per ml. 5 Cells were inoculated into 96-well cell culture plates at 200 μl per well, at 37°C with 5% CO 2 for 4 hours in an inc...
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Application Information
- IPC
- C07D207/44; A61K31/4015; A61P35/00; C12P17/10; C12R1/885
- CPC
- C07D207/456; C12P17/10
- Inventors
- 陈立; å¼ ç»´ç»´