A non-viral gene carrier material and its preparation method and application, brain-targeted gene transfer system and its preparation method and application

A delivery system, brain-targeting technology that can be used in other methods of inserting foreign genetic material, gene therapy, neurological diseases, etc., and can solve problems such as increased toxicity

Active Publication Date: 2015-09-16
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally speaking, the toxicity of PPI will increase sharply with the increase of generations, which is probably one of the reasons why PPI has not been studied as widely as PAMAM so far.

Method used

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  • A non-viral gene carrier material and its preparation method and application, brain-targeted gene transfer system and its preparation method and application
  • A non-viral gene carrier material and its preparation method and application, brain-targeted gene transfer system and its preparation method and application
  • A non-viral gene carrier material and its preparation method and application, brain-targeted gene transfer system and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0092] In the preparation method of the brain-targeted gene delivery system, step (2) is to oxidize lactoferrin with sodium periodate, and the chemical reaction formula is provided as follows.

[0093]

[0094] Step (3) is to use sodium cyanoborohydride to reduce the reaction solution of polyglutamic acid-polypropyleneimine and oxidized lactoferrin, and the chemical reaction formula is provided as follows.

[0095]

[0096] There are specific lactoferrin (Lf) receptors on the blood-brain barrier, and the transmembrane transfer function mediated by Lf through the blood-brain barrier has also been confirmed, and studies have shown that the endogenous Lf concentration in plasma is far from It is lower than the amount of Lf receptors present on the blood-brain barrier, therefore, the Lf selected in the present invention is a good brain targeting ligand.

[0097] Koppu et al. (J.Control Release,2010,143(2):215-221) linked the third generation PPI to transferrin (Tf), so that ...

Embodiment 1

[0100] A method for preparing a non-viral gene carrier material, comprising the following steps:

[0101] (1) The 2.0th generation polypropyleneimine (m value is equal to 2) (0.03g, 0.0427mmol) with 1,4-butanediamine as the core shown in formula B, benzyl L-glutamate N- Carboxylic anhydride (1.06g, 4.27mmol) and chloroform (90ml) were added to a 250ml single-necked flask, and stirred at 30°C for 72h. Evaporate most of the solvent under reduced pressure, add ether (200ml) at -20°C, stir for a while, let stand, filter the precipitate obtained after standing, wash with appropriate amount of ether, dry in vacuum at room temperature for 2 hours, and branched polybenzyl glutamate (white powder, 0.718g, yield 88.3%).

[0102] figure 1 For the branched polybenzyl glutamate that the embodiment of the present invention makes 1 H NMR chart, from figure 1 It can be seen in:

[0103] 1 H NMR (CDCl 3 ,500MHz): δ (ppm):

[0104] 1.95-2.15 (s, CHC H 2 CH 2 ),2.45(s,CHCH 2 C H 2 ...

Embodiment 2

[0119] A method for preparing a non-viral gene carrier material, comprising the following steps:

[0120] (1) Prepare branched polybenzyl glutamate according to the method of embodiment one step (1);

[0121] (2) The 2.0th generation polypropyleneimine (m value is equal to 2) (1.0g, 1.293mmol) with 1'4-butanediamine as the core shown in formula B, 2-hydroxypyridine (0.0769g), Anhydrous dimethyl sulfoxide (5ml) was added to a 25ml single-necked flask, and the branched polybenzyl glutamate (0.032g, 1.684×10 -3 mmol), stirred at 50°C for 48h. Add 10ml of deionized water, stir for 5min, transfer the reaction solution into a 3.5KDa dialysis bag, dialyze for 48h, and freeze-dry to obtain non-viral gene carrier material (light yellow powder, 0.050g).

[0122] The non-viral gene carrier material has the structure shown in formula B' (m value is equal to 2) as the center structure, the amino group at the end of the second polypropyleneimine (PPI) and the branched polybenzyl glutamate...

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PUM

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Abstract

The invention provides a non-viral genetic vector material and a preparation method and application thereof. Biodegradable golyglutamic acid is used as a skeleton of the non-viral genetic vector material and is connected with a poly(propyleneimine) dendrimer rich in amino groups; the non-viral genetic vector material has the characteristics of low toxicity, a high transfection rate, biodegradability and good biocompatibility and can be used for preparation of a treatment instrument for treating primary degenerative diseases of a central nervous system. The invention further provides a brain-targeted gene transfer system prepared through coupling of the non-viral genetic vector material with lactoferrins and a preparation method and application thereof. The brain-targeted gene transfer system can be used for preparation of the treatment instrument for treating primary degenerative diseases of the central nervous system.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a non-viral gene carrier material and its preparation method and application, a brain-targeted gene transfer system and its preparation method and application. Background technique [0002] RNA interference (RNA interference, RNAi) is a powerful and effective gene silencing tool, which has been widely used in research fields such as gene function, cell signal transduction pathway, drug target screening, and gene therapy. [0003] Alzheimer's disease (Alzheimer's disease, AD, Alzheimer's disease) is a primary degenerative disease of the central nervous system, the main clinical phase is dementia syndrome, and the disability and mortality rate is extremely high. With the development of gene therapy technology, it is a brand-new idea to use RNA interference to inhibit brain metabolic abnormalities and reduce the generation of β-amyloid (Aβ) in AD. But how to pass the small interfering ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G81/00C12N15/87A61K48/00A61P25/28
Inventor 鹏越峰尚鹏张璇蔡青青房丽晶余再丹
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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