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A kind of preparation method and application of polyionic micelle nanoparticle

A micellar nanometer and polyion technology, which is applied to medical preparations containing active ingredients, wave energy or particle radiation treatment materials, pharmaceutical formulations, etc., to reduce cytotoxicity, increase stability, and the preparation method is simple and easy to operate.

Inactive Publication Date: 2017-10-31
BEIJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the formation of polyion micelles is mostly formed by the interaction between polymers and polymers. CPPs are used as the components of polyion micelles, and they self-assemble with carboxymethyl chitosan to form nanomicelles, which do not involve Complicated chemical reactions and any toxic solvents, and the application as a carrier of water-soluble therapeutic reagents have not been reported yet

Method used

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  • A kind of preparation method and application of polyionic micelle nanoparticle
  • A kind of preparation method and application of polyionic micelle nanoparticle
  • A kind of preparation method and application of polyionic micelle nanoparticle

Examples

Experimental program
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Effect test

Embodiment 1

[0063] Embodiment 1 Preparation of Tat-carboxymethyl chitosan polyion micellar nanoparticles

[0064] (a) Precisely prepare a carboxymethyl chitosan (CMCS) solution with a concentration of 2.0 mg / ml and a Tat polypeptide solution with a concentration of 1.0 mg / ml by using a Tris-Hcl buffer solution with pH=7.4. Pass the CMCS solution and Tat solution through 0.22 μm microporous membrane respectively, and set aside. The CMCS solution was added into a 25mL round bottom flask, placed on a magnetic stirrer, and continuously stirred at a speed of 500r / min. At 25° C., 1 ml of the Tat solution was added dropwise to the above 2 ml of the CMCS solution at a rate of 1.0 ml / min. After the two solutions were mixed, the stirring was continued for 10 minutes at 25°C, and then left to stand for 20 minutes. Centrifuge the mixed solution for 10min at a centrifugal speed of 10000rpm, redissolve the obtained nanoparticles with 3ml Milli-Q ultrapure water, continue to centrifuge for 10min at a ...

Embodiment 2

[0071] Example 2 Preparation of nanomicelles loaded with miRNA

[0072](a) Precisely prepare a carboxymethyl chitosan (CMCS) solution with a concentration of 2.0 mg / ml and a Tat polypeptide solution with a concentration of 1.0 mg / ml by using a Tris-Hcl buffer solution with pH=7.4. Dissolve 20 μg of miRNA in 100 μL of RNase-free water and set aside. Pass the CMCS solution and Tat solution through 0.22 μm microporous membrane respectively, and set aside. The CMCS solution was added into a 25mL round bottom flask, placed on a magnetic stirrer, and continuously stirred at a speed of 500r / min. The solution for dissolving the miRNA was added to the CMCS solution. At 25° C., 1 ml of the Tat solution was added dropwise to the above 2 ml of the CMCS solution at a rate of 1.0 ml / min. After the two solutions were mixed, the stirring was continued for 10 minutes at 25°C, and then left to stand for 20 minutes. Centrifuge the mixed solution for 10min at a centrifugal speed of 10000rpm, ...

Embodiment 3

[0073] Example 3 Test of MiRNA Encapsulation Efficiency

[0074] The supernatant in the embodiment is analyzed, and the residual amount of miRNA in the supernatant is tested with a UV spectrophotometer at a wavelength of 260nm, compared with the input amount, and then the encapsulation efficiency of miRNA is calculated, and the encapsulation obtained by measuring is 92%.

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Abstract

The invention discloses a preparation method and application of polynano micelles. The nano particle is prepared by self-assembly of carboxymethyl chitosan and cationic polypeptide in buffer solution in the form of non-covalent bond. The preparation method of the nano-micelle particle of the present invention is simple, can be prepared in a buffer solution at room temperature, does not use any organic reagents and other harmful chemicals, the particle diameter is about 200nm, and the particle diameter distribution is uniform, and can be used in various solutions. Maintain stability for a long time. The nano micelles prepared by the invention can be used as carriers of water-soluble therapeutic reagents (photosensitizers, chemotherapeutic drugs, nucleic acids, etc.), and can improve the stability of the therapeutic reagents.

Description

technical field [0001] The invention belongs to the field of biomedical materials. It specifically relates to a preparation method of polyion micellar nanoparticles based on cationic cell penetrating peptide and carboxymethyl chitosan and its application as a water-soluble therapeutic agent carrier. Background technique [0002] Polyion complex micelles (PICMs) are dense particles formed by self-assembly of two or more oppositely charged polymers. Due to the hydrophilic segment, it can be stably dispersed in the water phase. PICMs are widely used in the preparation of nanoreactors, biosensors, and MRI agents. However, PICMs as carriers of therapeutic reagents (photosensitizers, enzymes, proteins, heparin, doxorubicin, and nucleic acids, etc.) are currently the hotspot of research. This is due to the following advantages of PICMs as carriers of therapeutic agents: (1) carrying charged therapeutic agents across biological environmental barriers, because PICMs have a hydrati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/42A61K47/36A61K9/107A61K41/00A61K45/00A61P35/00
Inventor 盛望胡克磊邓雄威张芳
Owner BEIJING UNIV OF TECH
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