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SiRNA delivery carrier and preparation method and application thereof

A delivery carrier and drug-carrying technology, which is applied in drug delivery, pharmaceutical formulation, liposome delivery, etc., can solve the problems of low yield of natural exosomes, complex components of exosomes, and difficulty in controlling the effect, and achieve low equipment prices , high encapsulation efficiency and easy operation

Pending Publication Date: 2021-09-24
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the yield of natural exosomes is very low, and the separation method is complicated, and the cost of obtaining exosomes on a large scale is high; and due to the complex composition of exosomes, many functions need to be studied, and it is difficult to control their effects

Method used

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  • SiRNA delivery carrier and preparation method and application thereof
  • SiRNA delivery carrier and preparation method and application thereof
  • SiRNA delivery carrier and preparation method and application thereof

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preparation example Construction

[0046] Another aspect of the embodiments of the present invention also provides a method for preparing the aforementioned siRNA delivery carrier, which includes:

[0047] Preparation of red blood cell membranes into nanovesicles;

[0048] siRNA and cationic liposomes are incubated to prepare siRNA / cationic liposome complexes;

[0049] And, the siRNA / cationic liposome complex is loaded in the nanovesicle to obtain the siRNA delivery carrier.

[0050] In some more specific embodiments, the preparation method includes: co-incubating siRNA and cationic liposomes for 20-30 min to prepare the siRNA / cationic liposome complex.

[0051] In some more specific embodiments, the preparation method includes: ultrasonically treating the siRNA / cationic liposome complex and nanovesicles for 1.0-15 min to obtain the siRNA delivery carrier.

[0052]In some more specific embodiments, the mass ratio of the siRNA, cationic liposomes and nanovesicles is 1: (3-5): (30-160).

[0053] In some more s...

Embodiment 1

[0129] (1) Add erythrocytes to 1×PBS, centrifuge at 800g at 4°C for 5 minutes, collect erythrocytes, and repeat twice. Then red blood cells were added to 0.25×PBS, incubated on ice for 1 hour with shaking, 14000 rpm, and centrifuged at 4°C for 10 minutes to collect red blood cell membranes. Then add 0.25×PBS, shake and incubate on ice for 1h, centrifuge at 14000rpm, 4°C for 10min, and collect the red blood cell membrane. Then the erythrocyte membrane aqueous solution, absolute ethanol and chloroform volume ratio are 1: 5: 10, stir, filter, rotary steam, dry, add water and ultrasonically disperse, prepare the erythrocyte membrane with better water dispersibility; Wherein, erythrocyte and 0.25× The volume ratio of PBS is 1:9.

[0130] (2) Put the collected erythrocyte membrane into an ultrasonic breaker for ultrasonic crushing, the conditions are 60W power, 25KHz frequency, horn model Φ2, ultrasonication for 1min. Then, through an Avantiminiextruder liposome extruder equipped ...

Embodiment 2

[0133] (1) Add erythrocytes to 1×PBS, centrifuge at 800g at 4°C for 5 minutes, collect erythrocytes, and repeat twice. Then red blood cells were added to 0.25×PBS, incubated overnight with shaking on ice, centrifuged at 14000 rpm, 4°C for 10 min, and the red blood cell membrane was collected. Then add 0.25×PBS, shake and incubate on ice for 1h, centrifuge at 12000rpm, 4°C for 20min, and collect the red blood cell membrane. Then, the volume ratio of erythrocyte membrane aqueous solution, absolute ethanol and chloroform is 1:5:10, stirred, filtered, rotary evaporated, dried, added with water and ultrasonically dispersed to prepare erythrocyte membrane with better water dispersibility. Wherein, the volume ratio of red blood cells and 0.25×PBS is 1:9.

[0134] (2) Place the collected erythrocyte membrane in an ultrasonic breaker for sonication, the conditions are 20W power, 20KHz frequency, horn model Φ2, ultrasonic 5min, and then use Avantiminiextruder liposome extruder, equippe...

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Abstract

The invention discloses an siRNA delivery carrier and a preparation method and application thereof. The siRNA delivery carrier comprises a nano vesicle and siRNA loaded in the nano vesicle, and the nano vesicle is prepared from an erythrocyte membrane. The invention further discloses an siRNA delivery carrier based on nucleic acid aptamer targeting, which comprises a nano vesicle, siRNA loaded in the nano vesicle and the cholesterol modified nucleic acid aptamer loaded on the surface of the nano vesicle. The nano vesicle is prepared from an erythrocyte membrane. In the siRNA delivery carrier based on nucleic acid aptamer targeting, which is prepared by the invention, the siRNA is loaded in the nano vesicle, the encapsulation efficiency is higher, and the siRNA delivery carrier is difficult to degrade.

Description

technical field [0001] The invention belongs to the technical field of targeted drug delivery, and in particular relates to an siRNA delivery carrier and its preparation method and application. Background technique [0002] Small interfering RNA, as a functional molecule for gene regulation, has been widely used in the treatment of various diseases, especially in the treatment of tumors. Small interfering RNA regulates the expression of tumor-related genes to inhibit tumor growth and promote tumor cell death, etc. However, how to accurately and efficiently deliver small interfering RNA to tumor sites has always been a key problem to be solved in this field. [0003] Although traditional small interfering RNA delivery methods such as cationic liposomes, dendrimers, cationic polymers, and inorganic nanoparticles can successfully deliver small interfering RNAs to cells, these carrier materials are difficult to meet the requirements for biocompatibility. sexual demands. In rec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/46A61K9/127A61K47/54A61K47/69A61K31/7088A61P35/00
CPCA61K47/46A61K9/1273A61K9/1277A61K47/545A61K47/554A61K47/6915A61K31/7088A61P35/00
Inventor 裴仁军吕海银
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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