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Preparation and application of amphiphilic polypeptide genetic vector

A technology of gene carrier and polypeptide, applied in the field of nanomedical material preparation, can solve problems such as difficult degradation, and achieve the effects of safety toxicity, high cell transfection efficiency and low cytotoxicity

Active Publication Date: 2018-10-09
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But whether it is ATRP or RAFT, the C-C bond in the product structure is very strong and not easy to break, so such a carrier is difficult to degrade when it enters the body

Method used

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  • Preparation and application of amphiphilic polypeptide genetic vector
  • Preparation and application of amphiphilic polypeptide genetic vector
  • Preparation and application of amphiphilic polypeptide genetic vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Drying of solvent

[0033] 500 mL THF and 1.0 g CaH 2 Added to a clean round bottom flask, stirred for 24 h and then distilled to obtain dry tetrahydrofuran. 500 mL of n-hexane and 1.0 g of CaH 2 Added to a clean round bottom flask, stirred for 24h and then distilled to obtain dry n-hexane. Mix 500 mL of DMF and 100 mL of toluene at 150 °C under N 2 Azeotropy occurs under protection, traces of water are evaporated, and when cooled to room temperature, dry DMF can be obtained.

[0034] (2) Preparation of CBZ-L-lysine-NCA monomer

[0035] 5.0 g of N-carbobenzyloxylysine (purchased from Sigma-Aldrich Company) and 5.0 g of triphosgene were dissolved in dry 100 mL of tetrahydrofuran, and magnetically stirred at 50°C until the solution became clear and transparent. Then the solution was distilled and concentrated to 50 mL, and N-benzyloxycarbonyl (CBZ)-protected N-benzyloxycarbonyl-α-L-amino acid-N-carboxylic acid anhydride (CBZ-L-lysine) was obtained by precipitatio...

Embodiment 2

[0047] (1) Drying of solvent

[0048] 500 mL THF and 1.0 g CaH 2 Added to a clean round bottom flask, stirred for 24 h and then distilled to obtain dry tetrahydrofuran. 500 mL of n-hexane and 1.0 g of CaH 2 Added to a clean round bottom flask, stirred for 24h and then distilled to obtain dry n-hexane. Mix 500 mL of DMF and 100 mL of toluene at 150 °C under N 2 Azeotropy occurs under protection, traces of water are evaporated, and when cooled to room temperature, dry DMF can be obtained.

[0049] (2) Preparation of CBZ-L-lysine-NCA monomer

[0050] 5.0 g of N-carbobenzyloxylysine (purchased from Sigma-Aldrich Company) and 5.0 g of triphosgene were dissolved in dry 100 mL of tetrahydrofuran, and magnetically stirred at 50°C until the solution became clear and transparent. Then the solution was distilled and concentrated to 50 mL, and precipitated with dry n-hexane to obtain the crude product of CBZ-L-lysine-NCA monomer. The crude product was then dissolved in 30 mL of tetr...

Embodiment 3

[0058] (1) Drying of solvent

[0059] 500 mL THF and 1.0 g CaH 2 Added to a clean round bottom flask, stirred for 24 h and distilled to obtain dry THF. 500 mL of n-hexane and 1.0 g of CaH 2 Add it to a clean round bottom flask, stir for 24 h and distill to obtain dry n-hexane. Mix 500 mL of DMF and 100 mL of toluene at 150 °C under N 2 Azeotropy occurs under protection, traces of water are evaporated, and when cooled to room temperature, dry DMF can be obtained.

[0060](2) Preparation of CBZ-L-lysine-NCA monomer

[0061] 5.0 g of N-carbobenzyloxylysine (purchased from Sigma-Aldrich Company) and 5.0 g of triphosgene were dissolved in dry 100 mL of tetrahydrofuran, and magnetically stirred at 50°C until the solution became clear and transparent. Then the solution was distilled and concentrated to 50 mL, and precipitated with dry n-hexane to obtain the crude product of CBZ-L-lysine-NCA monomer. The crude product was then dissolved in 30 mL of tetrahydrofuran and recrystall...

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Abstract

The invention discloses preparation and application of an amphiphilic polypeptide genetic vector, and belongs to the technical field of nanometer medical material preparation. The method comprises thefollowing steps of using hexamethyldisilazane as an initiator to initiate two kinds of polypeptide monomers of CBZ-L-lysine-NCA and L-leucine-NCA to perform NCA ring opening polymerization; finally removing CBZ protection groups to obtain an amphiphilic embedded polypeptide genetic vector of poly(L-lysine)50-b-poly(L-leucine)15. The polypeptide polymer provided by the invention has low toxicity andcan be used for compressing plasmid DNA to assemble regular nanometer particles; the transfection efficiency is high; the vector belongs to an effective genetic vector; in addition, the controllablepolymerization is used by the synthesis method; natural polypeptide is used as raw materials; the obtained polymers are safe and efficient.

Description

technical field [0001] The invention relates to a novel amphiphilic polypolypeptide gene carrier and its application, and belongs to the technical field of nanomedical material preparation. Background technique [0002] With the continuous deepening of cancer treatment research, nucleic acid has begun to become an important class of therapeutic agents. Nucleic acids primarily include plasmid DNA (pDNA), small (or short) interfering RNA (siRNA), microRNA (miRNA), messenger RNA (mRNA), and oligodeoxynucleotides (ODN). Gene vector transfection efficiency and safety are major concerns. In order to improve transfection efficiency, scientists are continuously optimizing the structure and properties of cationic polymer carriers, such as: carrier molecular weight, carrier charge density, hydrophobic segment of carrier, cell penetrating molecular ability and targeted therapy. Unfortunately, very little work has succeeded in unifying increased transfection efficiency and reduced tox...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G69/48C08G69/10C12N15/87
CPCC08G69/10C08G69/48C12N15/87
Inventor 张颖周志平
Owner JIANGSU UNIV
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