A biomimetic intelligent drug carrier for ischemic stroke targeting erythrocyte membrane and its preparation method
A technology for ischemic stroke and red blood cell membranes, applied in the field of medicine, can solve the problems of low targeting efficiency, achieve the effects of improving curative effect, prolonging circulation time, and avoiding phagocytosis
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Embodiment 1
[0032] Preparation and characterization of nanovesicles;
[0033] Prepared by self-assembly method, weigh 10mg PHB-Dextran dissolved in a mixed solvent of formamide and methanol (V:V=1:1) as the organic phase, weigh 1mg NR2B9C dissolved in 0.2M Tris-HCl buffer, and mix with organic After the phases were mixed, they were dispersed dropwise into 10ml of 0.5% poloxamer 188 aqueous solution under constant stirring (500rpm, 37°C), and continued to stir for 1-2h. μm microporous membrane to obtain the nanovesicle carrier (NP / NR2B9C), its morphology was characterized by transmission electron microscopy, see figure 2 right. The figure shows that the nanovesicle carrier has a regular spherical appearance, uniform size, and a particle size of about 165 nm when observed under a transmission electron microscope. The results of measuring the particle size and zeta potential of nanovesicles by laser particle size analyzer are shown in image 3 .
Embodiment 2
[0035] SHp-PEG 2000 - Preparation and characterization of DSPE;
[0036] TakeMal-PEG 2000 -DSPE (maleimide-polyethylene glycol 2000-phospholipid complex) 10mg, dissolved in 1ml N,N-dimethylformamide, another SHp10mg, dissolved in 1ml phosphate buffer (pH 7.0) . The two were sequentially added dropwise to 10ml of phosphate buffer (pH 7.0), under nitrogen protection, and magnetically stirred overnight to obtain the primary product. The product was then dialyzed (molecular weight cut-off 3 KDa) to remove excess SHp. After lyophilization, SHp-PEG was obtained 2000 -DSPE. Nuclear magnetic resonance instrument detects its NMR spectrum, see figure 1 , where A is the NMR spectrum of Mal-PEG2000-DSPE, B is SHp-PEG 2000 -The nuclear magnetic spectrum of DSPE, as can be seen from the figure, A figure shows that there is a maleimide characteristic peak at 6.7ppm, while the peak disappears in B figure, showing that Mal-PEG 2000- The maleimide group in DSPE has reacted with SHp, SHp...
Embodiment 3
[0038] Extraction of erythrocyte membrane and modification of cerebral ischemia homing peptide SHp;
[0039] The blood is extracted by low-osmotic extraction method, and the blood is collected in a container containing anticoagulant. In order to avoid the protease contained or adsorbed on the membrane from changing the membrane protein, the following operations should be performed at a low temperature of 4°C. Take the fresh blood of the rat and centrifuge it in a refrigerated centrifuge at 5000r / min for 10min, discard the supernatant and the fluffy precipitate deposited on the surface of the red blood cells. Wash 3 times with pre-cooled PBS (pH 7.4) equivalent to 3 times the hematocrit (4°C, 5000r / min, 15min). Add pre-cooled 0.01mol / L Tric-HCl (pH7.4) and mix with red blood cells (V:V=40:1), and place at 4°C for 1-2h. Then centrifuge at 9000r / min for 15min, discard the supernatant (repeat 3-5 times) until there are no red blood cells visible to the naked eye. Finally, white ...
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