Application of Dibenzyl Phosphate Citrate in Inhibiting Calcium Ion Deposition Diseases
A technology of citric acid and dibenzyl phosphate, applied in the field of chemical biology, can solve the problems of poor fat solubility and low stability
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Embodiment 1
[0012] Example 1. Preparation of tribenzyl citrate.
[0013] Take by weighing 28.0g of citric acid with a water of crystallization, put it in a 100ml three-necked bottle, then weigh 70.0g of benzyl alcohol and pour it into the three-necked bottle, connect the water separator, reflux device and stirrer, and place it at 160- Stir and react in an oil bath at 180°C for 8 hours, then distill off excess benzyl alcohol under reduced pressure in an oil bath at 185°C, and finally dissolve the oily substance in absolute ethanol under heat, and white crystals will precipitate after standing for several hours, then filter with suction. Wash the crystals with a small amount of absolute ethanol and dry to obtain pure tribenzyl citrate C 27 h 26 o 7 , Weighing 62.7g, the yield is 93%. The technical index of gained tribenzyl citrate is as follows:
[0014] White needle crystal m.p.50-51℃.
[0015] 1H NMR (CDCl3), δ: 2.86(d, J=12,2H), 2.94(d, J=12,2H), 5.07(s,2H), 5.10(S,4H), 7.25~7.35(m...
Embodiment 2
[0017] Example 2. Preparation of dibenzyl phosphate citrate.
[0018] Weigh 30.0 g of the above-prepared tribenzyl citrate, dissolve it in 25 ml of refined benzene, add it together to a 100 ml three-necked bottle equipped with a thermometer and a stirrer, stir and cool to 4 ° C under ice bath, and Under the protection of the liquid sampler, the 13.6gPCl dissolved in the liquid sampler 5 100ml of benzene solution was slowly added dropwise into the solution, the temperature was maintained at 3-5°C, after 2 hours of dropping, the drop was completed, and then stirred for 0.5h, under the premise of ensuring that the temperature did not rise (maintained at about 4°C), further Slowly add 4.85ml of distilled water dropwise, after dropping, a white precipitate will form, then stir for 1 hour, and finally filter with suction, wash with a small amount of benzene for 2-3 times, and then drain in KOH\NaOH environment to obtain white needle-like crystals Dibenzyl Phosphate Citrate C 20 h...
Embodiment 3
[0021] Example 3. Inhibitory effect of dibenzyl phosphate citrate on C2C12 calcification in mouse myoblasts.
[0022] Establish cell model, its method is as follows: First, mouse myoblast C 2 C 12 Take 5×10 5 cells / ml were inoculated in 24-well plates, and the next day, serum-free medium was added and cultured in an incubator for 24 hours. On the third day, use 1mCi / ml45Ca 2+ The labeled medium was exchanged for cells, and then ATP was added at a final concentration of 1 mM. Negative controls were performed with cells not treated with ATP or β-glycerophosphate. After 48 or 72 hours, the medium was discarded, the cells were washed 5 times with low-temperature Hank's equilibrium solution, and 0.1N NaOH was added. Finally, the amount of radioactive substance in the cell lysate was determined, and the experiment was carried out three times, and the average value was calculated. The results are shown in Table 1.
[0023] Table 1: Dibenzyl phosphate citrate inhibits C 2 C ...
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