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Targeted multifunctional nanoparticles and preparation method and application thereof

A multi-functional, nanoparticle technology, applied in the field of biomedicine, can solve the problems of lack of specificity and precision of transfection sites, failure of plasmids to enter cells well, and inapplicability of tumor gene therapy, so as to facilitate the observation of tumors and reduce Toxic and side effects, easy to operate effect

Inactive Publication Date: 2019-02-19
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Most small animal gene transfection uses direct injection of plasmids into the tail vein, but this method does not have the specificity and precision of the transfection site, and the injected plasmids cannot enter the cells well, and are easily degraded, resulting in low transfection efficiency and other defects, so it is not suitable for gene therapy of tumors
With nanoparticles as carriers, due to the EPR effect of tumors, nanoparticles themselves have a tendency to accumulate and phagocytize at tumor sites, and are often used as drug sustained-release carriers, so plasmids can be entrapped in the inner water phase of nanoparticles, but At present, there are still two disadvantages in encapsulating the plasmid in the inner water phase of the nanoparticle. One is that during the process of encapsulation, the synthesis of the nanoparticle requires the use of a sonic shock instrument, which is a kind of damage to the plasmid and makes it open. Second, the release efficiency of plasmids entrapped in the inner aqueous phase will be much lower than those adsorbed on the surface after entering the cells

Method used

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  • Targeted multifunctional nanoparticles and preparation method and application thereof
  • Targeted multifunctional nanoparticles and preparation method and application thereof
  • Targeted multifunctional nanoparticles and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of targeted multifunctional nanoparticles

[0034] (1) Dissolve 50 mg of polylactic-co-glycolic acid (PLGA) with a molecular weight of 12,000 and 2 mg of heptacyanine fluorescent small molecule IR780 in 1 mL of dichloromethane, and then add 200 uL of doxorubic acid hydrochloride with a concentration of 15 g / L Specific solution, under the conditions of ultrasonic power of 135w, ultrasonic frequency of 20Hz, and ice bath conditions, it was ultrasonically treated with a sonoshock instrument. During the treatment, it was ultrasonicated once every 5s, and the ultrasonic time was 5s each time. A total of 16 times of ultrasonication were added. The 40g / L polyvinyl alcohol aqueous solution was again ultrasonically treated with a sonic shock instrument under the conditions of ultrasonic power of 135w and ultrasonic frequency of 20Hz. During the treatment, ultrasonic treatment was performed every 5s, each ultrasonic time was 5s, and a total of 16 ultrasonic waves were ...

Embodiment 2

[0041] Preparation of targeted multifunctional nanoparticles

[0042] (1) Dissolve 50 mg of polylactic acid-glycolic acid copolymer (PLGA) with a molecular weight of 7000 and 3 mg of heptacyanine fluorescent small molecule IR780 in 1 mL of dichloromethane, then add 25 uL of double-distilled water, and use ultrasonic power of 135w , Ultrasonic frequency is 20Hz, under the condition of ice bath, use sonic shock instrument to ultrasonically process, ultrasonically once every 5s, each ultrasonic time is 5s, total ultrasonic 14 times, then add 4mL polyvinyl alcohol aqueous solution with a concentration of 20g / L , under the conditions of ultrasonic power of 135w and ultrasonic frequency of 20Hz, ultrasonic treatment was carried out using a sonoseismic instrument. During the treatment, ultrasonic treatment was performed once every 5s, and the ultrasonic time was 5s each time. Aqueous propanol solution to obtain mixed solution I. Stir the mixed solution I at a speed of 100r / min for 2....

Embodiment 3

[0046] Preparation of targeted multifunctional nanoparticles

[0047] (1) Dissolve 50 mg of polylactic-co-glycolic acid (PLGA) with a molecular weight of 9000 and 0.5 mg of heptacyanine fluorescent small molecule IR780 in 1 mL of dichloromethane, then add 300 uL of perfluoro-n-pentane, and The power is 135w, the ultrasonic frequency is 20Hz, and the sonic shock instrument is used for ultrasonic treatment under the condition of ice bath. During the treatment, ultrasonic treatment is performed every 5s, and the ultrasonic time is 5s each time. Ethylene alcohol aqueous solution, again under the conditions of ultrasonic power of 135w and ultrasonic frequency of 20Hz, was ultrasonically treated with a sonoshock instrument. During the treatment, it was ultrasonicated once every 5s, and the ultrasonic time was 5s each time. After a total of 17 times of ultrasonication, 10mL of % isopropanol aqueous solution to obtain the mixed solution I, stir the mixed solution I at a speed of 40r / m...

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Abstract

The invention relates to targeted multifunctional nanoparticles and a preparation method and application thereof, and belongs to the technical field of biological medicines. The targeted multifunctional nanoparticles are prepared by an ultrasonic double-emulsification method, covalent binding is carried out by branching PEI and PLGA, PLGA nanoparticles which have negative charge originally are modified into kation nanoparticles with positive charge, plasmids can be adsorbed effectively, it ensures that the plasmid cannot drop due to physical effect in a circulation system, and after the plasmid enters cells, the possibility that the plasmid is degraded by endoenzyme is also reduced. The nanoparticles further contain IR780 which can target tumor cell mitochondria, meanwhile, by strong fluorescent light and photosensitizer characteristics of IR780, the nanoparticles become a great fluorescent and photoacoustic imaging medium, and thus, tumors can be convenient to observe. The nanoparticles can carry drug to facilitate combination with transgenic therapy, tumor cells are killed favorably, and furthermore, toxic and side effects of chemotherapy drugs can further be reduced. The preparation method of the targeted multifunctional nanoparticles is simple, is easy to operate, and is suitable for expanded production.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a targeted multifunctional nanoparticle and its preparation method and application. Background technique [0002] Most small animal gene transfection uses direct injection of plasmids into the tail vein, but this method does not have the specificity and precision of the transfection site, and the injected plasmids cannot enter the cells well, and are easily degraded, resulting in low transfection efficiency And other defects, so it is not suitable for gene therapy of tumors. With nanoparticles as carriers, due to the EPR effect of tumors, nanoparticles themselves have a tendency to accumulate and phagocytize at tumor sites, and are often used as drug sustained-release carriers, so plasmids can be entrapped in the inner water phase of nanoparticles, but At present, there are still two disadvantages in encapsulating the plasmid in the inner water phase of the nanop...

Claims

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Application Information

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IPC IPC(8): A61K49/22A61K49/00A61K48/00A61K9/51A61K47/34A61K47/22A61K31/704A61P35/00
CPCA61K9/5123A61K9/5146A61K9/5153A61K31/704A61K48/005A61K49/0032A61K49/0093A61K49/225A61P35/00
Inventor 郝兰王梦竹王志刚
Owner CHONGQING MEDICAL UNIVERSITY
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