Application of penilloenol E1 derived from Trichoderma citrinoviride in lung cancer
A technology of Trichoderma citrinum and lung cancer cells, which can be used in medical preparations containing active ingredients, pharmaceutical formulations, respiratory diseases, etc., can solve problems such as no drugs, and achieve the effect of significant anti-lung cancer activity.
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Embodiment 1
[0015] Example 1 Fermentative production and separation and purification of compounds
[0016] 1. Fermentation production
[0017] Fermentation culture of production bacteria: according to the conventional method of cultivating microorganisms, take Trichoderma auranthoderma ( Trichoderma citrinoviride. ) IBPT-4 (preserved in the China Center for Type Culture Collection on January 25, 2013, address: Wuhan University, Wuhan, deposit number: CCTCC NO: M 2013055) appropriate amount, inoculated on PDA solid slant medium, in Cultivate for 4 days in a 28°C incubator.
[0018] Get the Trichoderma auranthoderma ( Trichoderma citrinoviride. ) appropriate amount of IBPT-4, inoculated into 400mL culture medium [composition of culture medium (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, NaCl 6.0 constant volume] in a 1000mL Erlenmeyer flask, after 30 days of static culture at 28°C, mycelia and fermentati...
Embodiment 2
[0026] Example 2 Test of anti-tumor activity in vitro
[0027] 1. Experimental samples and experimental methods
[0028] Preparation of the test sample solution: the test sample is the pure product of the compound isolated and refined in Example 1. Accurately weigh an appropriate amount of sample, and prepare a solution with the required concentration with methanol for activity measurement.
[0029] The subculture of cell lines and cells adopts lung cancer cell lines, and the cells use RPMI-1640 medium containing 10% FBS at 37°C and 5% CO 2 Subculture in an incubator.
[0030] Cell Proliferation Inhibitory Activity Test Method
[0031] Tetrazolium salt (MTT) method: Take cells in the logarithmic growth phase and adjust the cell density to 2×10 per ml 5 Cells were inoculated in 96-well cell culture plates at 200 microliters per well, and kept at 37°C in 5% CO 2 incubator for 4 hours. Add 2 microliters of sample solution or blank solution to each well, and after 24 hours o...
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