Sialyloligosaccharide-gold nano particle and preparation method and applications thereof
A technology of gold nanoparticles and sialic acid oligosaccharides, which is applied in the field of sialic acid oligosaccharides-gold nanoparticles and its preparation and application, can solve the problems of high cost, popularization and application, and inability to achieve good practical results
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Embodiment 1
[0048] Embodiment 1, the preparation of sialotriose-short connecting chain
[0049]
[0050] 1. Preparation of short connecting chains
[0051] Take 1.68 grams of the compound shown in formula III (4.96mmol, see the literature for its synthesis method: K.Kim, H.Yang, S.Jon, E.Kim, J.Kwak, J.Am.Chem.Soc.2004,126, 15368–15369.) were dissolved in 20 mL of dichloromethane (DCM) and 1 mL of triethylamine (Et 3 N, 7.5 mmol) and 780 μl of methanesulfonyl chloride (MsCl, 10 mmol); the reaction was stirred at room temperature for 1 hour. The reaction progress was monitored by TLC. After the raw materials disappeared, the reaction solution was filtered through diatomaceous earth and concentrated to obtain an orange-yellow slurry. Place the concentrate in 20 mL of acetonitrile (CH 3 CN), add 3.25 g of sodium azide (NaN 3 , 50mmol), reflux and stir overnight. After the reaction was detected by TLC, the solution was concentrated and evaporated to remove the solvent under reduced pr...
Embodiment 2
[0067] Embodiment 2, the preparation of sialotriose-short connecting chain
[0068]
[0069] Preparation of sialotriose-short linker chains
[0070] Take 14.87 mg of lactose short linker chain (formula II-1-1, final concentration of 10 mM) in Example 1, and 14.72 mg of CMP-N-acetylneuraminic acid (CMP-Neu5Ac, final concentration of 15 mM) dissolved in 2 ml After adding Tris-HCl (pH8.0) buffer solution, the temperature of the reaction system was raised to 37°C, and then 100 microliters of Photobacterium damsela α2,6-sialyltransferase (Photobacterium damselaα2,6-sialyltransferase, Pd2, 6-ST, 0.2U, purchased from Tianjin Suntech Pharmaceutical Science and Technology Co., Ltd.), after 2-3 hours, thin layer chromatography (TLC) detected that the raw material had completely disappeared, and the reaction solution was passed through an ultrafiltration membrane to remove enzyme protein, freeze-dried and concentrated After purification by gel column G-15, 15.5 mg of sialotriose-shor...
Embodiment 3
[0074] Embodiment 3, the preparation of sialotriose-short connecting chain
[0075]
[0076] 1. Synthesis of short linker chains of lactosamine
[0077] Take 60 mg of lactosamine propargyl glycoside (0.14 mmol) shown in formula I-2, see literature for its synthesis method: R.Daly, G.Vaz, A.M.Davies, M.O.Senge, E.M.Scanlan, Chem.Eur.J.2012, 18, 14671–14679.) and 77.63 mg of short linker chains (as shown in Example 1, formula IV, 0.214 mmol) were dissolved in 7 ml of N,N-dimethylformamide (DMF) and methanol (MeOH) (volume ratio 1:1), then add 13.3 mg cuprous iodide (CuI, 0.07 mmol), and react overnight under nitrogen protection. The disappearance of raw materials was monitored by TLC, the reaction solution was filtered, lyophilized and concentrated, and then purified by G-15 gel column to obtain a white solid which was the compound shown in formula II-1-2 (65.88 mg, yield 60%). Concrete reaction formula is as follows:
[0078]
[0079] The structural characterization da...
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