A novel hybrid nano calcium phosphate gene delivery system and its preparation method

A nano-calcium phosphate and gene delivery technology, applied in other methods of inserting foreign genetic materials, gene therapy, pharmaceutical formulations, etc., can solve the problems of complicated preparation methods and unfavorable expansion of production, and achieve simple and controllable process, convenient and reliable use , good safety effect

Active Publication Date: 2016-02-17
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this preparation method is relatively complicated and uses potentially toxic organic solvents and surfactants, which is not conducive to expanding production

Method used

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  • A novel hybrid nano calcium phosphate gene delivery system and its preparation method
  • A novel hybrid nano calcium phosphate gene delivery system and its preparation method
  • A novel hybrid nano calcium phosphate gene delivery system and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Preparation of a novel hybrid nano calcium phosphate gene delivery system.

[0058] 1) Preparation of reagent a, containing CaCl 2 250mM, Tris 20mM aqueous solution, hydrochloric acid to adjust the pH to 7.4;

[0059] 2) Preparation of reagent b, containing PEG-grafted carboxymethyl chitosan at a concentration of 200mg / L, Na 2 HPO 4An aqueous solution with a concentration of 1.5mM, a NaCl concentration of 280mM, and a Hepes concentration of 50mM, with NaOH adjusting the pH to 7.4;

[0060] 3) Mix 40 μg of gene DNA molecules with 1 mL of reagent, and quickly mix with reagent b evenly to prepare a gene transfection carrier of hybrid nano-calcium phosphate.

Embodiment 2

[0061] Example 2: Preparation of a novel hybrid nano calcium phosphate gene delivery system.

[0062] 1) Preparation of reagent a, containing CaCl 2 250mM, Tris 20mM aqueous solution, hydrochloric acid to adjust the pH to 7.4;

[0063] 2) Preparation of reagent b, containing PEG-grafted carboxymethyl chitosan at a concentration of 200mg / L, Na 2 HPO 4 An aqueous solution with a concentration of 1.5mM, a NaCl concentration of 280mM, and a Hepes concentration of 50mM, with NaOH adjusting the pH to 7.4;

[0064] 3) Take 120 μg gene DNA molecule and mix it with 1mL reagent, and then quickly mix it with reagent b evenly to prepare the gene transfection carrier of hybrid nano-calcium phosphate.

Embodiment 3

[0065] Example 3: Preparation of a novel hybrid nano calcium phosphate gene delivery system.

[0066] 1) Preparation of reagent a, containing CaCl 2 250mM, Tris 20mM aqueous solution, hydrochloric acid to adjust the pH to 7.4;

[0067] 2) Preparation of reagent b, containing PEG-grafted carboxymethyl chitosan at a concentration of 200mg / L, Na 2 HPO 4 An aqueous solution with a concentration of 1.5mM, a NaCl concentration of 280mM, and a Hepes concentration of 50mM, with NaOH adjusting the pH to 7.4;

[0068] 3) Mix 80 μg of gene DNA molecules with 1 mL of reagent, and quickly mix with reagent b evenly to prepare a gene transfection carrier of hybrid nano-calcium phosphate.

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Abstract

The invention discloses a novel hybrid nanometer calcium phosphate gene delivery system and a preparation method thereof. The gene delivery system comprises calcium phosphate and PEG (polyethylene glycol) grafted carboxymethyl chitosan, wherein a core is formed by wrapping a gene with calcium phosphate, the surface of the core is coated with PEG grafted carboxymethyl chitosan, and a hydrophilic shell is formed by a PEG chain segment, thus nano-particles with pegylation core-shell structures and particle sizes of 60-100nm are formed. The novel hybrid nanometer calcium phosphate gene delivery system disclosed by the invention has the advantages that organic solvents and surface catalysts do not need to be used in a preparation process, the process is simple and controllable, and the novel hybrid nanometer calcium phosphate gene delivery system is low in economic cost, good in repeatability and suitable for large-scale production and has low toxicity and high transfection efficiency. In addition, the invention also relates to a kit for preparing the gene delivery system and an application of the gene delivery system in in-vitro and in-vivo gene transfection and gene therapy.

Description

technical field [0001] The present invention relates to the field of pharmaceutical preparations and the field of biomedicine technology, in particular to a novel hybrid nano-calcium phosphate gene delivery system and a preparation method thereof. Background technique [0002] With the development of genetic engineering, nanotechnology, modern medical technology and other disciplines, treating diseases at the genetic level is becoming a very promising treatment. Gene therapy is to regulate the expression of key proteins involved in the occurrence and development of diseases by introducing nucleic acids (DNA, siRNA, miRNA) into tissues and cells, so as to achieve the purpose of disease treatment. However, due to its large molecular weight, negative charge, and hydrophilic characteristics, nucleic acid molecules cannot effectively enter cells; in addition, nucleic acid molecules are easily degraded by nucleases in plasma proteins. Therefore, nucleic acid molecules need to be ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/87A61K48/00
Inventor 孙敏捷谢颖张灿平其能张斐然
Owner CHINA PHARM UNIV
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