Nucleic acid aptamer delivery vector and preparation method and application thereof

A nucleic acid aptamer and delivery carrier technology, applied in the field of molecular biology, can solve the problems of complex separation methods, high cost, and low yield of natural exosomes, and achieve high biocompatibility, easy operation, and low equipment prices Effect

Pending Publication Date: 2021-01-05
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of natural exosomes is very low, and the isolation method i

Method used

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  • Nucleic acid aptamer delivery vector and preparation method and application thereof
  • Nucleic acid aptamer delivery vector and preparation method and application thereof
  • Nucleic acid aptamer delivery vector and preparation method and application thereof

Examples

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Example Embodiment

[0071]The embodiment of the present invention provides a small nucleic acid delivery vector based on aptamer targeting, its preparation method and its drug loading method, including:

[0072]Use hypotonic treatment to prepare red blood cells into red blood cell membranes;

[0073]Prepare the red blood cell membrane into nanovesicles by squeezing the membrane;

[0074]The cholesterol-modified aptamer, the cholesterol-modified small nucleic acid and the chemotherapeutic drug doxorubicin are loaded onto the membrane of the nanovesicle by incubation to prepare targeted drug-loaded nanovesicles.

[0075]The red blood cells are placed in a PBS solution and incubated for 1 to 3 hours with shaking, and centrifuged at 12000 to 14000 rpm for 10 to 20 minutes to collect the red blood cell membrane. The red blood cell membrane is ultrasonically broken, and then squeezed through the membrane 10-30 times to obtain nano-sized vesicles.

[0076]The ultrasonic breaker is used, the power is 20-60W, the frequency is...

Example Embodiment

[0118]Example 1

[0119]Step 1: Add the red blood cells to 1×PBS, centrifuge at 800g for 5 min at 4°C, collect the red blood cells, and repeat twice. Then the red blood cells were added to 0.25×PBS, and incubated on ice for 1 h, centrifuged at 14000 rpm, 4°C for 10 min, and the red blood cell membranes were collected. Then add 0.25×PBS, incubate with shaking on ice for 1 h, centrifuge at 14000 rpm, 4°C for 10 min, and collect the red blood cell membrane.

[0120]Among them, the volume ratio of red blood cells to 0.25×PBS is 1:9.

[0121]Step 2: Place the collected erythrocyte membrane in an ultrasonic disruptor and ultrasonically disrupt it under the conditions of 60W power, 25KHz frequency, horn model Φ2, and ultrasonic for 1min. Then, using the Avanti miniextruder liposome extruder, equipped with a 200nm pore size polycarbonate membrane, the erythrocyte membrane after ultrasound was squeezed through the membrane 30 times to obtain nanovesicles.

[0122]The above steps 1 to 2 can be passedfigu...

Example Embodiment

[0124]Example 2

[0125]Step 1: Add the red blood cells to 1×PBS, centrifuge at 800g for 5 min at 4°C, collect the red blood cells, and repeat twice. Then, the red blood cells were added to 0.25×PBS, and incubated on ice for 3 hours with shaking at 14000 rpm, and centrifuged at 4°C for 10 minutes to collect the red blood cell membranes. Then add 0.25×PBS, incubate with shaking on ice for 1 h, centrifuge at 12000 rpm, 4°C for 20 min, and collect the red blood cell membrane.

[0126]Among them, the volume ratio of red blood cells to 0.25×PBS is 1:9.

[0127]Step 2: Place the collected erythrocyte membrane in an ultrasonic disruptor and ultrasonically disrupt it under the conditions of 20W power, 20KHz frequency, horn model Φ2, and ultrasonic for 5min. Then use the Avanti miniextruder liposome extruder, equipped with a 100nm pore size polycarbonate membrane, and squeeze the erythrocyte membrane after ultrasound 10 times across the membrane to obtain nanovesicles.

[0128]The above steps 1 to 2 can...

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Abstract

The invention discloses a nucleic acid aptamer delivery carrier, a pharmaceutical composition containing the carrier, and a preparation method and application of the carrier and pharmaceutical composition. The nucleic acid aptamer delivery carrier comprises a nano vesicle taking an erythrocyte membrane as a skeleton, and the nano vesicle is loaded with a cholesterol-modified nucleic acid aptamer.The obtained drug-loaded targeting nano vesicle has the specificity to tumor cells, can silence the expression of P-glycoprotein in drug-resistant tumor cells and reverse the drug resistance of the drug-resistant tumor cells, and at the same time realizes the effective killing of the drug-resistant tumor cells at a relatively low adriamycin concentration.

Description

technical field [0001] The invention relates to a nucleic acid aptamer, in particular to a nucleic acid aptamer delivery carrier, and its preparation method and application belong to the field of molecular biology. Background technique [0002] Nucleic acid aptamers, also known as nucleic acid aptamers, aptamers, etc., are single-stranded oligos that are screened from artificially synthesized DNA / RNA libraries and can bind to various targets with high affinity and high specificity. Nucleotides. It was first proposed by the two groups of Szostak and Gold almost at the same time. In 1990, Ellington and Szostak reported an RNA fragment that could bind small molecule organic dyes and named it Aptamer. Aptamer (aptamer) refers to the systemic evolution of ligands by exponential enrichment (Systematic Evolution of Ligands by Exponential Enrichment, SELEX) technology, which is screened from artificially synthesized oligonucleotide libraries with high affinity and specificity. St...

Claims

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Application Information

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IPC IPC(8): A61K47/26A61K9/127A61K47/46A61K47/28A61K31/704A61K31/7105A61P35/00
CPCA61K47/26A61K9/1273A61K47/46A61K47/28A61K31/704A61K31/7105A61P35/00A61K2300/00
Inventor 裴仁军王腾飞
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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