Chalcone compounds and uses of the composition
A technology of chalcones and compounds, applied in the field of chalcones
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preparation Embodiment 1
[0098] hematoxychalcone
[0099] Take 100 g of Sapphire medicinal material, extract with 1000 ml of 95% ethanol under heat reflux for 1 hour, and extract the dregs with 1000 ml of 95% ethanol once more. Combine the above two extracts, concentrate the extract to 100ml of water, extract with 100ml of ethyl acetate, and repeat the above extraction steps once more. The ethyl acetate layer was removed and concentrated to dryness to obtain a crude product. Take 10g of the crude product, mix the sample in 30g of 100-200 mesh silica gel, carry out normal phase separation on a silica gel column (400g, 200-300 mesh), and use the following ratios of chloroform:methanol mixed solution (200:1, 100:1, 50:1) , 25:1, 10:1) 5000ml gradient elution, and finally washed with methanol. A total of 116 fractions were collected. According to the TLC detection results, the fractions were combined, and Fr.62-90 was purified by repeated silica gel column chromatography to obtain an orange powder compo...
Embodiment 1
[0107] Effect of Hematoxychalcone on Microtubule Polymerization
[0108] Experimental instrument: GENios Pro multifunctional microplate reader, purchased from Tecan Company.
[0109] Experimental reagents: Tubulin Polymerization Assay Kit, purchased from Cytoskeleton Company (Cat.#CDS03andBK006).
[0110] Experimental method (subject to the instructions of the tubulin polymerization kit):
[0111] 1. Preheat the 96-well plate at 37°C for 30 minutes;
[0112] 2. Pre-cool tubulin polymerization buffer at 4°C:
[0113] General Tubulin Buffer (750 μl): 80 mM piperazine-N, N'-bis(2-ethanesulfonic acid) (PIPES) (pH6.9), 2 mM Mgcl2, 0.5 mM ethylene glycol bis (2-aminoethyl ether) tetraacetic acid (EGTA);
[0114] Tubulin Glycerol Buffer (250 μl): 15% glycerol (glycerol) was dissolved in general-purpose tubulin buffer;
[0115] Guanosine triphosphate (GTP Stock) (200mM, 10μl): 1mM guanosine triphosphate (GTP);
[0116] 3. Preheat 500μl General Tubulin Buffer to room temperature;...
Embodiment 2
[0124] Effects of Hematoxychalcone on Cell Cycle of Human Gastric Carcinoma HGC
[0125] Culture HGC cells in vitro, when the growth state is good, digest and inoculate 1×10 5 Cells were placed in a 6cm culture dish and adhered to the wall overnight. After the cells are in good condition, suck off the culture medium, add the hematoxychalcone (formula II) prepared in Preparation Example 1 diluted with the culture medium, the final concentration is 2 μg / ml, add the culture medium to the negative control well, and keep at 37°C Cultivate for 24h. Changes in cell morphology were observed under a microscope. The cells were digested with trypsin (0.25% trypsin / 0.02% EDTA), gently blown down, transferred to a centrifuge tube, centrifuged at 2000rpm for 5min, washed twice with PBS, and centrifuged at 1000rpm for 5min. Slowly add 800 μl of 75% ethanol (pre-cooled at -20°C) to the cell pellet, and shake (Vortex) while dripping to make it form a single cell as much as possible. After ...
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