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Preparation and application of nano-composite system of polycation macromolecular prodrug used for gene transfer

A cationic polymer and macromolecular technology, applied in the biomedical field of nanomaterials, can solve the problems of non-guidance, complex preparation, immunogenicity and safety hazards, and achieve good stability, good biocompatibility, and normal cytotoxicity small effect

Inactive Publication Date: 2019-04-05
NANHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Viral vectors have the mechanism of congenitally infecting cells and are the main means of transfection. However, viral vectors have limited packaging capacity, complex preparation, immunogenicity (cannot be used repeatedly in vivo), potential safety hazards, and non-guiding

Method used

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  • Preparation and application of nano-composite system of polycation macromolecular prodrug used for gene transfer
  • Preparation and application of nano-composite system of polycation macromolecular prodrug used for gene transfer
  • Preparation and application of nano-composite system of polycation macromolecular prodrug used for gene transfer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] GC-FUA used in the construction of microRNA-122 delivery liver targeting nanosystem

[0081] Weigh LA1.1g and dissolve in 25mL TEMED / HCl buffer solution (pH4.7), add 0.3g EDC·HCl, stir magnetically for 20min to activate, then add 0.07gNHS and 1.1g chitosan, stir magnetically at room temperature After reacting for 72 hours, put it into a dialysis bag for dialysis for 2 days, and freeze-dry to obtain a light yellow solid sample GC.

[0082] In a three-necked flask equipped with a reflux condensing device, add 10.0 g of KOH and 30 mL of double-distilled water, and stir to dissolve. Then add 4.0 g of 5-Fu, keep stirring in a 40°C water bath for 30 min. Then slowly add 6.25g of bromoacetic acid solution 10mL dropwise over 30min. Slowly warm up to 60°C, and react at this temperature for 6h. Cool to room temperature, adjust the pH to 5.5 with concentrated HCl, put it in the refrigerator, and refrigerate overnight. If there is precipitation, filter it off, collect the filtra...

Embodiment 2

[0085] Construction of a liver tumor-targeting nanosystem for p53-delivered macromolecular prodrug GC-FUA-FA

[0086] (1) The implementation of GC-FUA synthesis is the same as in Example 1.

[0087] (2) Preparation of GC-FUA-FA: Weigh 0.5 g of galactosylated chitosan, dissolve it in 5 ml of acetic acid-sodium acetate buffer, slowly add a DMSO solution of folic acid active ester under magnetic stirring conditions, Protect from light for 16 hours, then adjust the pH to 8.0-10.0 with NaOH. The mixture was dialyzed in distilled aqueous solution for 7 days. Finally freeze-dried yellow powder GC-FUA-FA.

[0088] (3) The characterization method and transfection method are the same as in Example 1.

Embodiment 3

[0090] Construction of a liver tumor-targeting nanosystem for nucleic acid aptamer delivery using dual-target GC-FA-DOX

[0091] (1) GC synthesis is the same as in Example 1.

[0092] (2) Dissolve 500mg of FA in 25ml of DMSO, stir until completely dissolved, add 0.4g of EDC, continue to stir, add GC solution dissolved in acetic acid-sodium acetate in advance, stir for 1 day in the dark, dialyze for 48h, freeze-dry to obtain GC-FA . A 2 mg / ml GC-FA solution was prepared with acetic acid solution, TPP and DOX solutions were pre-prepared and mixed, and the mixed solution of DOX and TPP was added dropwise to the GC-FA solution. Stir for 24h, dialyze for 48h, and rotary evaporate to obtain GC-FA-DOX.

[0093] The characterization method and transfection method are the same as in Example 1.

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Abstract

The invention discloses a preparation and application of a nano-composite system of a polycation macromolecular prodrug used for gene targeted transfer. The composite system comprises a high polymer material, such as polycation, a targeted group, an antitumor drug and a target gene. According to the preparation and application of nano-composite system disclosed by the invention, the advantages ofnanometer drug delivery system and macromolecular prodrug are integrated; the antitumor drug and the target gene are coupled to polycation through chemical bonds so as to compound the macromolecular prodrug, and then a self-assembling method is adopted for preparing a macromolecular prodrug nano-composite system; the target gene is coated during a self-assembling process, the target gene is effectively protected and the gene is prevented from enzymatic degradation. According to the preparation and application of nano-composite system disclosed by the invention, the polycation macromolecular prodrug serves as a gene vector, co-transfer of drug and gene is realized, advantages of gene therapy and chemotherapy are integrated and the nano-composite system has an excellent application prospectin the aspect of oncotherapy.

Description

technical field [0001] The invention belongs to the field of nanomaterial biomedicine. The invention relates to a targeted delivery gene / drug nanocomposite, in particular to a preparation method and application of a nanocomposite of a macromolecular prodrug used for targeted treatment of tumors to encapsulate a tumor suppressor functional gene. Background technique [0002] Macromolecular prodrugs refer to compounds in which low-molecular drugs are linked to suitable macromolecules through covalent bonds, and the macromolecular prodrugs are enzymatically hydrolyzed or hydrolyzed to release active drugs to exert their medicinal effects. Macromolecular prodrugs can prolong the release time of drugs, reduce the dosage, and reduce toxic and side effects; they can increase the water solubility of drugs with low solubility or insoluble in water, improve the kinetics of drugs, and reduce multidrug resistance. . [0003] Gene therapy is an important field of modern medical researc...

Claims

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Application Information

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IPC IPC(8): A61K47/54A61K47/59A61K47/61A61K48/00A61K31/704A61K31/513A61P35/00
CPCA61K47/549A61K31/513A61K31/704A61K47/545A61K47/59A61K47/61A61K48/005A61P35/00
Inventor 喻翠云宁倩
Owner NANHUA UNIV
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