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A multifunctional nano-biological material transfection reagent with the effect of gene therapy for Parkinson's disease, its preparation method and application

A technology for Parkinson's disease and biomaterials, applied in the field of multifunctional nano-biomaterial transfection reagents and its preparation, can solve the problems of Parkinson's disease and non-existing nano-biomaterials, and achieve the improvement and treatment of Parkinson's disease symptoms, less side effects, and improved curative effect

Active Publication Date: 2016-06-29
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A multifunctional nano-biological material transfection reagent with the effect of gene therapy for Parkinson's disease, its preparation method and application
  • A multifunctional nano-biological material transfection reagent with the effect of gene therapy for Parkinson's disease, its preparation method and application
  • A multifunctional nano-biological material transfection reagent with the effect of gene therapy for Parkinson's disease, its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 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 to 26 mL In the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 50 h in an ice bath at 4°C and in the dark;

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

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

[0073] S2. Preparat...

Embodiment 2

[0078] Embodiment 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 to 27 mL In the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 55 hours in an ice bath at 4°C and in the dark;

[0082] S12. After the synthesis is completed, use ultrafiltration centrifuge tubes (MiliporeMolecut 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), and freeze-dry spare;

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

[0084...

Embodiment 3

[0089] Embodiment 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 to 30 mL In the above-mentioned phosphate buffer solution (PBS solution, pH=7.4) containing N-succinimide ester and dimethylformamide (DMF), stir and react for 60 h in an ice bath at 4°C and in the dark;

[0093] S12. After the synthesis is completed, use ultrafiltration centrifuge tubes (MiliporeMolecut 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), and freeze-dry spare;

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

[0095] ...

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Abstract

The invention provides a multifunctional nano-biological material transfection reagent with the effect of gene therapy for Parkinson's disease and a preparation method thereof. The method is to immobilize nerve growth factor (NGF) on hydrogel-wrapped iron ferric oxide nanoparticles (Fe 3 o 4 -OA-NAPI-AA) surface, and then adsorb plasmid DNA that can interfere with α-synuclein synthesis. The nanomaterial transfection reagent acts on MPP + After induction, the apoptosis of PC12 cells, an in vitro Parkinson's disease model, was significantly inhibited, the expression of nerve growth factor receptor (NGFR) was up-regulated, the expression of α-synuclein was down-regulated, and the classic BAX, P53, Bcl-2 apoptosis pathway to achieve the purpose of treating Parkinson's disease. The nano material transfection reagent of the present invention is non-toxic, has few side effects, can successfully carry genes into cells, and has obvious effects, and is worthy of popularization and application in the treatment of Parkinson's disease.

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 was called "parkinsonism" in the early stage. Later, the disease was named Parkinson's disease (Parkinson's disease) and is still used 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-dopamine preparati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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