Application of gamma-dehydroharmine extracted and separated from peganum harmala to preparation of medicine for inhibiting gastric cancer
A technology for dehydrogenation of camellia alkaloids and camellia officinalis, which is applied in the directions of medical preparations, drug combinations, antitumor drugs and the like containing active ingredients
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Embodiment 1
[0016] Extraction and separation of γ-dehydrohalamine:
[0017] Preparation of the total alkaloids of camel succulents:
[0018] a. Get 5 kg of camel balsam medicinal material, add 8, 6 and 6 times the concentration of 70% ethanol aqueous solution to extract 3 times, each time for 1 hour, combine the extracts, concentrate at 60 ° C, and dry into a dry extract to obtain a total of 1375g;
[0019] b. Add 500g of the obtained extract, add 300mL of 10% sulfuric acid solution, dissolve the alkaloids in the extract in stages, filter, combine the extracts, add 25% ammonia solution to pH = 9.5, filter after standing for 24 hours, and precipitate Dry to obtain 11.696g of total alkaloid extract;
[0020] Preparation of sample solution:
[0021] Weigh 1g of the total alkaloid extract into a test tube, add 5mL of the upper phase with triethylamine added and 5mL of the lower phase without hydrochloric acid, and ultrasonically oscillate to dissolve them for separation by pH zone counterc...
Embodiment 2
[0029] CCK-8 method to detect the inhibitory effect of γ-dehydrohalamine on the proliferation of gastric cancer cells:
[0031] Take out the RPMI-1640 medium containing 10% FBS (fetal bovine serum) from the refrigerator at 4°C, place it in a water bath at 37°C to preheat for 20 minutes, wipe the outer bottle wall with 75% alcohol cotton balls after returning to temperature, and transfer it into The ultra-clean bench that has been sterilized by ultraviolet light is ready for use; take out the cell cryopreservation tube containing human gastric adenocarcinoma cell SGC-7901, immediately transfer it to a 37°C water bath for rapid thawing, and transfer the thawed cryopreservation tube into a centrifuge. Centrifuge at 800rpm for 5min, take out the cryopreservation tube, gently unscrew the cap of the cryopreservation tube, use a 1ml pipette gun to discard the upper cryopreservation solution, avoid sucking up the cells at the bottom of the tube, add 1ml RPMI...
Embodiment 3
[0039] Molecular docking analysis of the binding effect of γ-dehydrohalamine on gastric cancer-related targets:
[0040] In the present invention, firstly, 17 active ingredients (including γ-dehydrohalaline) of the national medicine camel are subjected to ligand pretreatment, and finally 14 active ingredients are obtained by screening, and the 14 active ingredients are combined with AKT1, TP53, JUN, MAPK1 Carry out molecular docking with EGFR to realize the interaction between the receptor and the ligand according to the principle of energy, and find the best binding mode;
[0041] Select the original ligand to be separated from the active pocket and carry out docking, and calculate the binding energy (Score) value after docking, the lower the binding energy (Score), the more stable the conformation of ligand and receptor binding, the interaction may occur The greater the resistance; the binding energy (Score) ≤ -4.25KJ / mol indicates that the ligand has a certain binding activ...
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