A kind of soluble thrombin nanoparticle and its preparation method and application
A nanoparticle and thrombin technology, applied in the field of biomedicine, can solve the problems of low solubility, high toxicity, low drug efficacy, etc., and achieve the effects of high bioavailability, good stability, and reduced drug dosage.
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Embodiment 1
[0045] Preparation of amylose-polylysine nanoparticles:
[0046] (1) Synthesis of the first-generation dendritic polylysine by amidation reaction: 1.73g L-2,6-di-tert-butoxycarbonyllysine (5mmol) and 350μL propargylamine (5.5mmol) were co-dissolved in 20mL In anhydrous N,N dimethylformamide (DMF) solution, stir under nitrogen for 10min, then cool to 0°C, add 2.18g (5.5mmol) benzotriazole-N,N,N to the solution ', N'-tetramethyluronium hexafluorophosphate (HBTU) and 0.74g (5.5mmol) 1-hydroxybenzotriazole (HOBT), continue to stir at room temperature for 16h, after the reaction, add 200mL ethyl acetate Ester (EtOAc), with saturated NaHCO 3 solution, 0.1mol / L NaHSO 4 , saturated NaHCO 3 Solution and brine were rinsed sequentially, and then dried by rotary evaporation. The obtained crude product was purified by column chromatography (silica gel, chloroform-methanol=5:1), and then rotary evaporation could obtain 90% of the first generation dendrimer lysine.
[0047] (2) Synthesi...
Embodiment 2
[0051] Take 20 mg of amylose-polylysine nanoparticles obtained in Example 1, put them into a three-neck flask, add 500 ml of acetic acid aqueous solution with a mass percentage of 2%, and stir to make it dissolve completely; company) 3000u, add phosphate buffer (pH7.4) 100ml, stir to dissolve, and form a thrombin buffer solution; stir at a speed of 100-800 rpm at room temperature, and add the thrombin buffer solution dropwise to the linear chain Into the starch-polylysine acetic acid aqueous solution, the dropping time is 30min, the temperature is raised to 40°C, 30ml of glutaraldehyde solution with a mass fraction of 0.1% is added dropwise, stirring continuously at a speed of 200r / min for 30min, the stirring is stopped, and poured Put it into 500ml of liquid paraffin, stir for 25min, add 50ml of Tween 80, and stir at a high speed of 500 rpm for 4 hours to emulsify; take the emulsion and add 100ml of glutaraldehyde solution with a mass fraction of 25%, and stir for 30min to mak...
Embodiment 3
[0053] Take 40 mg of amylose-polylysine nanoparticles obtained in Example 1, put them into a three-necked flask, add 1500 ml of acetic acid aqueous solution with a mass fraction of 2%, and stir to make it dissolve completely; company) 6000u, add 150ml of phosphate buffer (pH7.4), stir to dissolve, and form a thrombin buffer solution; stir at a speed of 100-800 rpm at room temperature, and add the thrombin buffer solution dropwise to the above linear Into the starch-polylysine acetic acid aqueous solution, the dropping time is 40min, the temperature is raised to 40°C, 40ml of glutaraldehyde solution with a mass fraction of 0.1% is added dropwise, and the stirring is continued for 40min at a speed of 300 rpm, the stirring is stopped, and it is poured Put it into 500ml of liquid paraffin, stir for 35min, add 100ml of Tween 80, and stir at a high speed of 500 rpm for 4 hours to emulsify; take the emulsion and add 150ml of glutaraldehyde solution with a mass fraction of 25%, and sti...
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