Targeted nano drug carrier modified by hyaluronic acid oligosaccharide based on electrostatic spraying and its preparation method and application
A technology of hyaluronic acid oligosaccharides and nano-drug carriers, which is applied in the field of electrostatic spraying and nano-materials, can solve the problems of poor drug slow release, uneven particle size distribution, toxic and side effects, etc., and achieve good slow-release effect and good drug Sustained-release, easy-to-operate effect
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Embodiment 1
[0051] Example 1: Preparation of targeted nano-drug carriers modified with hyaluronic acid oligosaccharides by coaxial electrostatic spraying method
[0052] The HA-CTS polymer was prepared with reference to the method in Example 1 in the patent application document "A hyaluronic acid oligosaccharide-chitosan microsphere loaded with docetaxel and its preparation method and application" (CN108670960A).
[0053] Prepare HA-CTS polymer by cross-linking hyaluronic acid oligosaccharide and chitosan with EDC / NHS, the specific method is as follows: add hyaluronic acid oligosaccharide 90mg, EDC 180mg, NHS 195mg to 30mL 0.05M MES buffer successively , each reagent was added after the previous reagent was completely dissolved. After magnetic stirring for 30 minutes, add 180 mg of CTS and continue stirring. React at room temperature for 20 hours, centrifuge at 5500 rpm / min for 5 minutes, collect the supernatant, and adjust the pH to about 7.10 with 1M NaOH. After continuing to react for ...
Embodiment 2
[0058] Example 2: Drug loading and encapsulation efficiency of targeted nano-drug carriers modified by hyaluronic acid oligosaccharides
[0059] Establishment of DTX standard curve:
[0060] Accurately weigh 2.0mg of DTX standard substance in a 10mL volumetric flask, dissolve it with methanol and dilute to the mark, prepare a standard solution with a concentration of 200μg / mL, and then serially dilute to 160, 120, 80, 40, 20, 10μg / mL series solutions, filtered with 0.22 μm filter membrane, HPLC detection and analysis chromatogram, with concentration (C) as the abscissa, peak area (A) as the ordinate to make a linear regression standard curve, the standard curve of DTX is as follows Figure 4 As shown, the regression equation obtained by simulation is: Y=11054X-27129(R 2 =0.9929), indicating that DTX has a good linear relationship in the concentration range of 10-200 μg / mL.
[0061] Weigh 0.4 mg of the hyaluronic acid oligosaccharide-modified targeted nano drug carrier prepa...
Embodiment 3
[0067] Example 3: In vitro release of targeted nano-drug carriers modified by hyaluronic acid oligosaccharides
[0068] Weigh 1 mg of the hyaluronic acid oligosaccharide-modified targeted nano-drug carrier prepared in Example 1 and add it to 6 mL of PBS buffer (40 g NaCl, 1 g KCl, 1.2 g K 2 HPO 4 , 7.2 g Na 2 HPO 4, add double-distilled water to make up to 1L) in a dialysis bag (Mw=30kDa), and perform dialysis in 400mL of PBS buffer solution with different pH, the magnetic stirrer rotates at 200rmp / min, and the temperature is 37°C. The pH of the PBS buffer simulating the internal environment of tumor cells is 6.8, and the pH of the PBS buffer simulating the physiological internal environment is 7.4. Sample 10 mL at 1, 2, 4, 8, 12, 24, 36, 48, 60, 72, 96, and 120 h, and add an equal volume of PBS buffer with the same pH. After the sample was lyophilized, it was reconstituted with 1 mL of double-distilled water and then extracted with 1.4 mL of ethyl acetate. Take 1 mL of th...
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