Unlock instant, AI-driven research and patent intelligence for your innovation.

Polyamide-amine type branch-shape polymer nano materials, synthesis method and use thereof

A nanomaterial and dendritic technology, applied in the field of materials science, can solve the problems of difficulty in obtaining meaningful gene expression and low transduction efficiency

Active Publication Date: 2006-09-27
SHANGHAI BLOOD CENT
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is that in the prior art, vectors mediating gene transfer are generally divided into viral vectors and non-viral vectors. Viral vectors have defects such as inducing human tumors and immune rejection, while traditional non-viral vectors , although it is safe, the transduction efficiency is extremely low, and it is difficult to obtain meaningful gene expression. The technical solution adopted by the present invention to solve the above technical problems is to provide a polyamide-amine type dendritic polymer nanomaterial, which is a Monodisperse nanoparticles with highly branched spherical structure, molecular size of 50-90 nm

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Polyamide-amine type branch-shape polymer nano materials, synthesis method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Synthesis of G5 polyamide-amine dendrimers

[0019] 1. Pretreatment of raw materials

[0020] Methanol was added magnesium powder, refluxed for 4h and then purified by distillation. Add sodium hydroxide to ethylenediamine, let it stand for 24h, and purify by distillation. Methyl acrylate was added with anhydrous calcium chloride, refluxed for 4 hours and purified by distillation.

[0021] 2. Synthesis of G5 PAMAM dendrimer

[0022] Synthesis of generation 0.5 (G0.5): Take 0.7ml (10mmol) of ethylenediamine in a round bottom flask, add 20ml of methanol, slowly add 9.0ml (100mmol) of methyl acrylate dropwise, and stir electromagnetically at room temperature for 3d. After the reaction was completed, methanol and excess methyl acrylate were removed by rotary evaporation to obtain 3.8 g of light yellow viscous liquid. Yield 94.1%.

[0023] Synthesis of generation 1 (G1): Take 3.8 g of G0.5 obtained in the previous step, add 40 ml of methanol, slowly add 20 ml of ethylene...

Embodiment 2

[0025] Example 2 Gene transfection of hematopoietic stem cells

[0026] 1. Purification of Hematopoietic Stem Cells

[0027] The umbilical cord blood collected for no more than 4 hours was separated from mononuclear cells by Ficoll, and CD34-positive hematopoietic stem cells were separated by CD34 immunomagnetic beads, and the purity reached 95% by flow cytometry. Cultured for one day in serum-free medium containing hematopoietic stem cell growth factor.

[0028] 2. Gene Transfection of Hematopoietic Stem Cells

[0029] After the plasmid DNA containing the target gene is purified with an endotoxin-free plasmid extraction kit, it is diluted to 1 ug / ul with TE buffer, and the prepared nanoparticles are made into a 5 mg / ml aqueous solution, and the nanoparticles and plasmid DNA are mixed at a ratio of 5: 1, mixed evenly, added dropwise to the hematopoietic stem cell culture system, cultivated in a carbon dioxide incubator at 37°C for 2 hours, centrifuged and washed once, added ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to materialogical field, it has disclosed a polyamide-maine branch-typed polymer nm material. Use methyl acrylate and ethylenediamine as the original monomer, by synthesizing the fifth generation of PAMAM branch-typed high molecular (dendrimer), use dialysis to purify the product, generating monodispersed nm particle with branch-typed ball structure, the diameter of molecule is 50-90nm with super high density of positive functional group and cavity inside. Being mixed with DNA, the particles can transmit the goal gene, the protein synthesized in which can be expressed for a long time in hematopoietic stem cell, to the hematopoietic stem cell purified by CD34 immune megnatic ball.

Description

technical field [0001] The invention belongs to the field of materials science, and relates to a nano material and its preparation and application. Further, the invention relates to a polyamide-amine type dendritic polymer nano material, its synthesis method and application. Background technique [0002] The rise and development of gene therapy has brought more effective treatments for various human diseases, especially for various malignant tumors, single-gene genetic diseases, and various refractory and lethal diseases. disease. However, there are still many obstacles to be overcome for the clinical application of gene therapy. Among them, the carrier problem, as well as the carrier-related immune response, cytotoxicity and safety issues are one of the most critical issues to be solved in the field of gene therapy research. Vectors that mediate gene transfer are generally divided into viral vectors and non-viral vectors. Viral vectors have defects such as inducing human...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C08G69/00
Inventor 范华骅陆华中郑滨高峰
Owner SHANGHAI BLOOD CENT