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Multifunctional nano-biomaterial transfection reagent with effect of treating parkinsonism by gene, as well as preparation method and application thereof

A technology for Parkinson's disease and biomaterials, which is applied in the field of multifunctional nano-biomaterial transfection reagents and its preparation, can solve the problems of Parkinson's disease that have not been seen, and have no nano-biomaterials, and achieve small side effects, improvement and The effect of treating Parkinson's disease and improving the curative effect

Active Publication Date: 2014-09-17
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant reports, and there is no research on the combination of targeted drug delivery of nano-biological materials combined with gene regulation and its application in the treatment of Parkinson's disease.

Method used

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  • Multifunctional nano-biomaterial transfection reagent with effect of treating parkinsonism by gene, as well as preparation method and application thereof
  • Multifunctional nano-biomaterial transfection reagent with effect of treating parkinsonism by gene, as well as preparation method and application thereof
  • Multifunctional nano-biomaterial transfection reagent with effect of treating parkinsonism by gene, as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Preparation of multifunctional nano-biological material transfection reagent of the present invention

[0068] Proceed as follows:

[0069] S1. Preparation of photoactive NGF

[0070] S11. According to the volume ratio of 1:4, add dimethylformamide (DMF) to phosphate buffer solution (PBS solution, pH=7.4), then add 800 μg N-succinimide ester; add 60 μg NGF In 26mL of the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 50h in an ice bath at 4°C and in the dark;

[0071] S12. After the synthesis is completed, use ultrafiltration centrifuge tubes (Milipore Molecut Ⅱ, 10KNa) to centrifuge for 30 minutes at a speed of 4000 rpm / min to purify the generated azidophenyl derivatives, which are photoactive NGF (AzPhNGF). freeze-dried for later use;

[0072] S13. Before use, add 50 mL of PBS solution to dissolve and adjust to the desired concentration of 1 ng / μL.

[0073]...

Embodiment 2

[0078] Example 2 Preparation of multifunctional nano-biological material transfection reagent of the present invention

[0079] Proceed as follows:

[0080] S1. Preparation of photoactive NGF

[0081] S11. According to the volume ratio of 1:5, add dimethylformamide (DMF) to phosphate buffer solution (PBS solution, pH=7.4), then add 800 μg N-succinimide ester; add 60 μg NGF In 27mL of the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 55h in an ice bath at 4°C and in the dark;

[0082] S12. After the synthesis, use ultrafiltration centrifuge tubes (Milipore Molecut Ⅱ, 10KNa) to centrifuge for 40 minutes at a speed of 3500 rpm / min to purify the generated azidophenyl derivatives, which are photoactive NGF (AzPhNGF). freeze-dried for later use;

[0083] S13. Before use, add 50 mL of PBS solution to dissolve and adjust to the desired concentration of 1 ng / μL.

[0084] S2. Preparat...

Embodiment 3

[0089] Example 3 Preparation of multifunctional nano-biological material transfection reagent of the present invention

[0090] Proceed as follows:

[0091] S1. Preparation of photoactive NGF

[0092] S11. According to the volume ratio of 1:4.5, add dimethylformamide (DMF) to phosphate buffer solution (PBS solution, pH=7.4), then add 850 μg N-succinimide ester; add 60 μg NGF In 30mL of the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 60h in an ice bath at 4°C and in the dark;

[0093] S12. After the synthesis is completed, use ultrafiltration centrifuge tubes (Milipore Molecut Ⅱ, 10KNa) to centrifuge for 30 minutes at a speed of 4500 rpm / min to purify the generated azidophenyl derivatives, which are photoactive NGF (AzPhNGF). freeze-dried for later use;

[0094] S13. Before use, add 50 mL of PBS solution to dissolve and adjust to the desired concentration of 1 ng / μL.

[009...

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Abstract

The invention provides a multifunctional nano-biomaterial transfection reagent with an effect of treating parkinsonism by gene, as well as a preparation method and application thereof. According to the method, a nerve growth factor (NGF) is immobilized on the surface of a hydrogel coated ferroferric oxide nano-particle (Fe3O4-OA-NAPI-AA) by virtue of a photo-grafting method, and plasmid DNA (Deoxyribonucleic Acid) capable of interfering with alpha-synuclein synthesis is adsorbed. The nano-material transfection reagent acts on the cell of an MPP+ (1-methyl-4-phenylpyridinium) induced in-vitro parkinsonism model PC12, and cell apoptosis is remarkably inhibited, wherein expression of the nerve growth factor receptor (NGFR) is up-regulated, expression of alpha-synuclein is down-regulated, and classic apoptosis pathway of BAX, P53 and Bcl-2 is influenced so as to achieve the aim of treating parkinsonism. The nano-material transfection reagent is non-toxic and less in side effects, can smoothly carry a gene to enter cells, is remarkable in action, and is worthy of popularization and application of parkinsonism treatment.

Description

technical field [0001] The invention belongs to the field of nano biological materials. More specifically, it relates to a multifunctional nano-biological material transfection reagent with the effect of gene therapy for Parkinson's disease, its preparation method and application. Background technique [0002] Parkinson's disease (PD), a common middle-aged and elderly nervous system degenerative disease, was first described by the British doctor James Parkinson (James Parkinson) in 1817, and it was called "parkinson's paralysis" in the early stage. The disease was later named Parkinson's disease (Parkinson's disease) and is still in use today. As the name implies, "shaking paralysis" is due to the slow movement of the patient after the illness, tremors in the hands or feet and other parts of the body, loss of flexibility of the body, and muscle stiffness. [0003] At present, the main treatment for Parkinson's disease is drug therapy, the most important of which is L-dopam...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P25/16A61K38/18
Inventor 关燕清刘俊明武迪
Owner SOUTH CHINA NORMAL UNIVERSITY
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