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MiRNA delivery carrier based on ultra-small gold nano-particles as well as preparation method and application of miRNA delivery carrier

A nanoparticle and delivery carrier technology, applied in biochemical equipment and methods, recombinant DNA technology, medical preparations with non-active ingredients, etc., can solve the problems of serious side reactions, high reaction temperature, long-term toxicity, etc. , high controllability, the effect of promoting osteogenic differentiation

Active Publication Date: 2017-02-22
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the general gold nanoparticles used for gene delivery larger than 10nm will accumulate in different tissues, and it is difficult to clear them through the kidneys, which may cause long-term toxicity (Longmire, M.; Choyke, P.L. et al. Nanomedicine2008, 3, 703-717 ; Wang, B.; He, X.; et al. Accounts of chemical research 2013, 46, 761-769. Karmali, P.P.; Simberg, D. Expert opinion on drug delivery 2011, 8, 343-357)
The current method of using gold nanoparticles to prepare miRNA delivery carriers is the chemical grafting method, but this method has the disadvantages of high reaction temperature, long time and serious side reactions.

Method used

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  • MiRNA delivery carrier based on ultra-small gold nano-particles as well as preparation method and application of miRNA delivery carrier
  • MiRNA delivery carrier based on ultra-small gold nano-particles as well as preparation method and application of miRNA delivery carrier
  • MiRNA delivery carrier based on ultra-small gold nano-particles as well as preparation method and application of miRNA delivery carrier

Examples

Experimental program
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Effect test

Embodiment 1

[0041] 1) Preparation of ultra-small gold nanoparticles: add chloroauric acid and citric acid to deionized water according to the molar ratio of 1:0.9, stir vigorously and quickly add the freshly prepared sodium borohydride solution dropwise to obtain the reaction solution, continue to stir the reaction After 5 hours, the reaction system was centrifuged and resuspended in water to obtain citric acid-capped ultra-small gold nanoparticles with a size of about 8 nm. Wherein the sodium borohydride in the sodium borohydride solution that adds and the mol ratio of chloroauric acid are 22.5:1, and the concentration of chloroauric acid in the reaction solution is 4mmol / L, and the concentration of citric acid is 3.6mmol / L, sodium borohydride The concentration is 0.09mol / L.

[0042] 2) Modification of ultra-small gold nanoparticles: 0.04 mmol of ultra-small gold nanoparticles prepared in step 1) were dissolved in 15 mL of sodium hydroxide solution at pH=10, and 0.036 mmol of 16-mercapto...

Embodiment 2

[0046] 1) Preparation of ultra-small gold nanoparticles: Add chloroauric acid and citric acid into deionized water according to the molar ratio of 0.2:0.2, stir vigorously and quickly add the freshly prepared sodium borohydride solution dropwise to obtain the reaction solution, and continue to stir the reaction After 6 hours of reaction, the reaction system was centrifuged and resuspended in water to obtain citric acid-capped ultra-small gold nanoparticles with a size of about 9 nm. Wherein the sodium borohydride in the sodium borohydride solution that adds and the mol ratio of chloroauric acid are 12:0.6, and the concentration of chloroauric acid in the reaction solution is 2mmol / L, and the concentration of citric acid is 2mmol / L, the sodium borohydride The concentration is 0.04mol / L.

[0047] 2) Modification of ultra-small gold nanoparticles: 0.01 mmol of ultra-small gold nanoparticles prepared in step 1) were dissolved in 20 mL of sodium hydroxide solution at pH=9, and 0.01...

Embodiment 3

[0051] 1) Preparation of ultra-small gold nanoparticles: Add chloroauric acid and citric acid into deionized water according to the molar ratio of 0.625:1, stir vigorously and quickly add the freshly prepared sodium borohydride solution dropwise to obtain the reaction solution, continue to stir the reaction After 8 hours of reaction, the reaction system was centrifuged and resuspended in water to obtain citric acid-capped ultra-small gold nanoparticles with a size of about 10 nm. Wherein the sodium borohydride in the sodium borohydride solution that adds and the mol ratio of chloroauric acid are 30:1.25, and the concentration of chloroauric acid in the reaction solution is 5mmol / L, and the concentration of citric acid is 8mmol / L, the sodium borohydride The concentration is 0.12mol / L.

[0052] 2) Modification of ultra-small gold nanoparticles: 0.02 mmol of ultra-small gold nanoparticles prepared in step 1) were dissolved in 25 mL of sodium hydroxide solution at pH=11, and 0.02 ...

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Abstract

The invention provides a miRNA delivery carrier based on ultra-small gold nano-particles as well as a preparation method and application of the miRNA delivery carrier. By utilizing layer-by-layer assembling method, polymine or lipidosome is assembled to the surfaces of the ultra-small gold nano-particles, or polymine and lipidosome are sequentially assembled to the surfaces of the ultra-small gold nano-particles from inside to outside, so as to obtain the miRNA delivery carrier based on the ultra-small gold nano-particles. The preparation method is convenient, simple and high in controllability, the stability of lipidosome can be improved, the toxicity of PEI can be reduced, and the superiorities of the ultra-small gold nano-particles can be adequately utilized. The particle sizes of the gold nano-particles in the carrier are less than 10nm, so that the renal toxicity is avoided; the carrier has good cytocompatibility, so that the delivery efficiency of miRNA can be improved; the carrier can be endocytosed by cells after loading miRNA, so that the osteogenic differentiation of mesenchymal stem cells from bone marrow can be promoted.

Description

technical field [0001] The invention belongs to the field of biotechnology of miRNA delivery carrier, and specifically relates to a miRNA delivery carrier obtained by surface modification of ultra-small (less than 10 nm) gold nanoparticles. Osteogenic differentiation of stem cells. Background technique [0002] As functional biomolecules, miRNAs are often used for bone tissue defect regeneration and disease treatment (Stegen, S.; van Gastel, N. et al. Bone 2015,70,19-27; Li, Y.; Fan, L . et al. Biomaterials 2013, 34, 5048-58; Zhang, X.; Li, Y. et al. Nat. Commun. 2016, 7). Many results have shown that miRNA can be delivered to tissue regeneration areas and target cells. [0003] However, since miRNAs and cell membranes both have the same type of charge—negative charges—it is difficult for them to cross cell membranes on their own. Delivered miRNAs are often degraded very quickly in vivo (Kai, Z.S.; Pasquinelli, A.E. et al. Nat. Struct. Mol. Biol. 2010, 17, 5-10). Therefo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N5/077A61K47/02A61K47/34A61K9/127A61K9/10A61K31/7105A61K48/00A61P19/08
CPCA61K47/02A61K47/34A61K9/10A61K9/127A61K31/7105C12N5/0654C12N15/85C12N2501/65C12N2506/1353
Inventor 雷波于萌
Owner XI AN JIAOTONG UNIV
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