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Cerebral arterial thrombosis targeted red cell membrane bionic intellectual drug carrier and preparation method thereof

A technology for ischemic stroke and red blood cell membrane, applied in the field of medicine, can solve problems such as low targeting efficiency, and achieve the effects of improving curative effect, prolonging circulation time, and avoiding phagocytosis

Active Publication Date: 2017-01-04
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the unmodified red blood cell membrane biomimetic carrier (RBC-NP) has certain passive targeting properties, its targeting efficiency is low, especially in the treatment of stroke. An ideal cerebral ischemia-targeted drug delivery system must Therapeutic drugs are further targeted and enriched in the lesion in the brain, increasing the drug concentration in the lesion in the brain, improving the curative effect, and reducing the impact on the normal physiological function of the brain

Method used

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  • Cerebral arterial thrombosis targeted red cell membrane bionic intellectual drug carrier and preparation method thereof
  • Cerebral arterial thrombosis targeted red cell membrane bionic intellectual drug carrier and preparation method thereof
  • Cerebral arterial thrombosis targeted red cell membrane bionic intellectual drug carrier and preparation method thereof

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Effect test

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 initial 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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Abstract

The invention discloses a cerebral arterial thrombosis targeted red cell membrane bionic intellectual drug carrier. The drug carrier is prepared from a nanometer vesica, a red cell membrane and cerebral ischemia homing peptide SHp, wherein the nanometer vesica serves as an inner core and is used for packing NR2B9C, the red cell membrane is arranged on the outer layer, the cerebral ischemia homing peptide SHp is modified on the surface of the cerebral ischemia, and the wrapping fusion between the nanometer vesica serving as the inner core and the red cell membrane on the outer layer is achieved through an ultrasonic fusion technique. The nanometer vesica is prepared from amphiphilic block copolymer (PHB-Dextran) serving as a carrier material, the amphiphilic block copolymer is prepared from hydrophobic borate which is grafted to a hydrophilic glucan framework, the nanometer vesica is used for wrapping a neuroprotective agent NR2B9C, and the amino acid sequence of the neuroprotective agent NR2B9C is KLSSIESDV. The carrier has the characteristics of having biocompatibility, prolonging circulation time in the body and initiatively targeting the focus position of the cerebral arterial thrombosis.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to an ischemic stroke-targeted red blood cell membrane bionic intelligent drug carrier and a preparation method thereof. Background technique [0002] Stroke is a neurological disease with high mortality, disability and recurrence, and is listed as the third killer of human beings after cardiovascular diseases and malignant tumors. Clinically, about 87% of strokes are ischemic strokes. Therefore, research on the drug treatment of ischemic stroke has great clinical significance. Studies have shown that: compared with normal physiological conditions, the body can continuously produce high levels of reactive oxygen species (ROS) at high levels in cerebral ischemic lesions under the pathological conditions of stroke, and use excessive ROS as the release switch to design Intelligent drug delivery system has great application prospects. NR2B9C is a polypeptide with great clinical pros...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/50A61K9/127A61K38/08A61K47/48A61K47/36A61P25/00A61P9/10
CPCA61K9/1273A61K9/5068A61K38/08A61K47/36
Inventor 辛洪亮徐群为吕伟江艳徐剑培
Owner NANJING MEDICAL UNIV
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