House lizard glycopeptide, method for preparing same and medicinal application thereof
A technology for shougong sugar and shougong sugar, which is applied in the field of shougong glycopeptide and its preparation, can solve the problems of toxicity, limit the dosage of drugs, influence on the treatment effect, etc., and achieve the effect of good biological activity and inhibiting tumor cell migration.
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Embodiment 1
[0040] Geckoglycopeptide, characterized in that: the weight average molecular weight is greater than 2,000,000Da; the specific rotation is -60°; the mass percentage of carbon element is 23.39%, the mass percentage of hydrogen element is 4.925%, and the mass percentage of nitrogen element The percentage content is 6.51%; the sugar part of its composition contains rhamnose, fucose, mannose, glucose, galactose, N-acetylglucosamine, and the molar ratio is: rhamnose: fucose: mannose : Glucose: Galactose: N-acetylglucosamine = 1.334: 3.363: 3.375: 6.983: 10.480: 0.019; the connection mode of monosaccharides is: 1,3-linked rhamnose, T-linked fucose, 1,2,3,6-,1,3,4-linked mannose, T-,1,4-linked glucose, T-,1,4-linked galactose and 1,4-linked N-acetylglucosamine; the mass percentage of uronic acid is 3.08%; its binding protein part contains 16 kinds of amino acids, and the mass percentage is 16.33%; the type of glycopeptide bond contains O-linkage; The mass percentage is 14.7%.
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Embodiment 2
[0043] Preparation of gecko glycopeptide: Take 1000g of the whole gecko, cut into 0.2cm long pieces, soak in 6000ml, 3000ml of methanol for 2h, filter, put the dregs in a ventilated place at room temperature to dry, and then use 8000ml, 6000ml , 4000ml of hot water was extracted three times at 100°C, and the extraction time was 6h, 4h, and 3h respectively. Combine the extracts, filter, concentrate to 800ml, add ethanol precipitation with a final concentration of 80% (V / V), centrifuge (3200r / min, 10min), collect the precipitate, redissolve the precipitate in water, centrifuge (3200r / min, 10min ), take the supernatant, remove free protein 10 times by conventional sevage method, concentrate, dialyze, concentrate, and dry. Take dry powder 24g and dissolve in 1200ml containing 0.5mol / L sodium chloride, 10mmol / L cysteine, 0.1mol / L phosphate buffer (pH6.5) of 10mmol / L EDTA, add 120mg Papain and 10ml of toluene, 60 ° C water bath for 48 hours, then add 60 mg of papain, continue to 60...
Embodiment 3
[0045] Preparation of Gecko Glycopeptide: Take 1000g of the whole Gecko, crush it into coarse powder, pass through a No. 1 sieve, soak in 3000ml of methanol for 12h, filter, put the dregs in a ventilated place at room temperature to dry, and then use 10000ml of hot water at 60°C Extract three times, the extraction time is 6h. Combine the extracts, filter, concentrate to 1000ml, add ethanol with a final concentration of 70% (V / V) to precipitate, stand at 4°C for 24h, centrifuge (4000r / min, 6min), collect the precipitate, redissolve the precipitate in water, and centrifuge (4000r / min, 6min), take the supernatant, dialyze and dry. Take dry powder 24g and dissolve in 4200ml containing 0.5mol / L sodium chloride, 10mmol / L cysteine, 0.1mol / L phosphate buffer (pH6.5) of 10mmol / L EDTA, add 480mg Papain and 40ml toluene, 70°C water bath for 8h, then add 240mg of papain, continue 70°C water bath for 4h. The enzymolysis solution was filtered, concentrated under reduced pressure to 500ml,...
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