Application of triptolide in preparation of antimalarial drugs, and pharmaceutical composition comprising triptolide
A technology of triptolide and its composition, which is applied in the field of carrier pharmaceutical compositions, and can solve the problems of antimalarial effect and drug resistance of triptolide that have not yet been seen
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Embodiment 1
[0020] Embodiment 1: triptolide extraction separation method
[0021] Extraction: Weigh 25kg of the whole herb of Tripterygium wilfordii, add 15L of 75% ethanol, and stir evenly. After the raw material is soaked for a set time, it is loaded into the extraction column, and the two ends of the extraction column are covered with medical gauze to prevent the filter of the extraction column from clogging and the fine particles of the material from entering the pipeline. Then hang the extraction column into the extraction kettle and seal it. Open the pipeline of the equipment to allow CO2 to enter the extraction kettle, adjust the pressure and temperature to the set value, close the pipeline of the extraction kettle, and keep it static for the set time, then open the pipeline and adjust the pressure and temperature of the separation kettle to the set value Value, dynamic cycle extraction. After the dynamic circulation extraction starts, the product is received from the split tank....
Embodiment 2
[0024] Example 2: Determination of triptolide antimalarial activity in vitro
[0025] Antimalarial activity can be determined by measuring the content of Plasmodium dsDNA (Corey, V.C., et al., A broad analysis of resistance development in the malaria parasite. Nat Commun, 2016.7: p.11901.). Add erythrocyte suspension (100 μL, cultured in RPMI1640) infected with 3D7 strain P. Add 5g / L AlbuMAX (Gibco) and 50mg / L Gentamicin (Sigma) to base (Gibco) to make malaria parasitemia reach 0.5%, hematocrit reach 2%), then place at 37°C, 5% Cultivate for 48 hours in a CO2 incubator. Cells were lysed with lysate (Tris 2.4g / L, EDTA 1.86g / L, Saponin 0.08g / L, TritonX-100 0.8mL / L) from Sigma, and dsDNA content was determined with 10000X SYBR Green I reagent (invitrogen) , the determination procedure refers to the literature (Plouffe, D., et al., In silico activity profiling reveals the mechanism of action of antimalarials discovered in a high-throughput screen. Proc Natl AcadSci U S A, 2008.1...
Embodiment 3
[0028] Example 3: In vitro toxicity test to mammalian cells
[0029]Carried out in 96-well tissue culture plates (J.Mustafa, S.I.Khan, G.Ma, L.A.Walker and I.A.Khan, Synthesis and Anticancer Activities of Fatty Acid Analogs of Podophyllotoxin.Lipids.2004,39(2):167-172 .). 25x10 in a 96-well plate 3 African green monkey kidney aneuploid cells (Vero) were planted at a density of one per well and incubated for 24 hours. Samples of different concentrations, ie, triptolide compound solutions, were added at concentrations of 1 μM, 3 μM, and 10 μM, and incubated for 48 hours. The number of surviving cells was determined by the Neutral Red assay method, and the IC was calculated from the dose-effect curve 50 . Doxorubicin was used as a positive control drug. Triptolide IC on Vero 50 The values are as follows:
[0030]
[0031] The results showed that triptolide had no obvious cytotoxicity at 1-10 μM in vitro.
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