Degradable polyamino acid derivative as well as preparation method and application thereof

A polyamino acid and biodegradation technology, used in the introduction of foreign genetic material using a carrier, recombinant DNA technology, etc., can solve problems such as high toxicity, and achieve the effects of strong pH buffering capacity, easy operation and high safety.

Inactive Publication Date: 2012-11-21
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its non-degradability and high toxicity inhibit its application in vivo.

Method used

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  • Degradable polyamino acid derivative as well as preparation method and application thereof
  • Degradable polyamino acid derivative as well as preparation method and application thereof
  • Degradable polyamino acid derivative as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 carrier material PSI-NN' x -NN y preparation of

[0030] Put 12.5 grams of DL-aspartic acid into a 500mL round-bottomed flask, mix evenly with 6.25 grams of 85% phosphoric acid, install the flask on a rotary evaporator, maintain a reduced pressure state and react in an oil bath at 180°C for 2.5 hours. The product was dissolved in 50 mL of N,N'-dimethylformamide, then 200 mL of water was added to precipitate the product, filtered and washed with water until neutral, and dried at 110°C for 24 hours.

[0031] Dissolve 1 gram of polysuccinimide in 15 mL of N,N'-dimethylformamide (DMF), add X mol of dimethyldipropylenetriamine and Y mol of 3,3'-diamino Dipropylamine (see Table 1 for the feed ratio, X+Y=24mmol), react in ice bath for 1 hour, dialyze with a dialysis membrane with a molecular weight cut-off of 14000 for 48 hours, and freeze-dry.

[0032] Determination of the ratio of primary, secondary and tertiary three amino groups in the polyamino acid deriva...

Embodiment 2

[0041]Embodiment 2 PSI-NN' x -NN y Determination of physical and chemical properties of

[0042] Determination of Carrier Material PSI-NN' by High Performance Gel Permeation Chromatography x -NN y To determine the molecular weight of its degradation products, water was used as the mobile phase, the flow rate was 0.8 ml / min, and the column temperature was 40°C. Use dextran (12000Da, 50000Da, 80000Da, 150000Da, 670000Da) as a reference substance, do a regression line with the logarithm of the retention time to the molecular weight, and PSI-NN' x -NN y Substitute the retention time into the above equation to obtain PSI-NN' x -NN y Molecular weight (see Table 1 for results).

[0043] Weigh 20mgPSI-NN’ x -NN y Materials, dissolved in 1mL of phosphate buffer solution, placed in a 37°C water bath for 1 to 30 days, and samples were taken to determine their molecular weight distribution by high performance gel permeation chromatography, the method is the same as above, and the...

Embodiment 3

[0045] Embodiment 3 PSI-NN' x -NN y Determination of binding ability to plasmid DNA

[0046] Configure a series of PSI-NN' x -NN y The mixture with DNA was allowed to stand still for 30 minutes, and was determined by agarose gel electrophoresis with a voltage of 120v, a current of 80mA, and an electrophoresis time of 40 minutes. See the experimental results Figure 4 . Figure 4 Medium PSI-NN’ x -NN y Agarose gel electrophoresis showed that they could bind plasmid DNA well.

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Abstract

The invention provides a degradable polyamino acid derivative which has a molecular weight of 30,000-40,000Da. A series of primary, secondary and tertiary amino groups in a certain proportion are introduced to a side chain so that the degradable polyamino acid derivative has very strong pH buffering capability and higher capability of transferring and expressing a plasma DNA; and the DNA binding capability, the transfection efficiency and the cytotoxicity can be adjusted and optimized through adjusting the proportion of the primary, secondary and tertiary amino groups. The synthesizing steps are simple and easy to operate, and the safety is high; the structural frame of the degradable polyamino acid derivative is polyamino acid which has the advantages of degradability, low toxicity and the like, can be applied to preparation of a degradable polycation nano gene carrier material and has a structural formula as the specification.

Description

technical field [0001] The invention belongs to the field of biological materials, and in particular relates to a series of derivatives of biodegradable polyamino acid materials that efficiently transfer and express genes, and their application in the preparation of degradable polycation nanometer gene carrier materials. technical background [0002] Gene therapy achieves the purpose of treatment by introducing normal genes into target cells to treat congenital or acquired genetic defects. In gene therapy, if naked DNA is directly injected into target tissues or cells, the transfection efficiency is often very low. This is because the uptake rate of exogenous DNA by cells is very low, and exogenous DNA is easily cleared by organisms or even degraded by itself. Therefore, a safe and efficient gene carrier has become an essential part of gene therapy. Although the virus vector has high efficiency, it has great security risks. Non-viral gene vectors have many advantages, suc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G69/48C08G69/10C08G69/08C12N15/63
Inventor 汤谷平马楠李文忠王维巍赵丹军周峻古斯塔夫.施坦因霍夫
Owner ZHEJIANG UNIV
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