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Method for carrying drugs or nano particles into specific target cells based on aptamer and cell-penetrating peptide

A nucleic acid aptamer and nanoparticle technology, applied in the field of nano-biomedical materials, can solve the problems of lack of cell specificity, poor stability of cell-penetrating peptides, limited application value, etc., to enhance targeting and enhance biological phase. Capacitive, method-simple effect

Inactive Publication Date: 2017-05-17
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the poor stability of cell-penetrating peptides and the lack of cell specificity limit their potential application value to some extent.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Polyethylene glycol-modified distearoylphosphatidylethanolamine (DSPE-PEG 2000-Mal) and cysteine-modified cell-penetrating peptide TAT (Cys-TAT) were dissolved at a ratio of 1:1.5 in triethylamine-containing chloroform / methanol (v / v = 2:1) solution and incubated at 25°C for 24 hr in the dark. Excess organic solvent was then removed by vacuum drying, and the precipitate was redissolved in chloroform. After filtering the solution with a filter membrane with a pore size of 0.02 μm, the solution was vacuum-dried again to remove the organic solvent, and the resulting precipitate (DSPE-PEG 2000-TAT) was stored at -20°C until use.

[0024] Weigh 11.5 mg of soybean lecithin (SPC), 0.96 mg of cholesterol, and 1.15 mg of DSPE-PEG 2000-TAT, dissolve the above materials in chloroform, and after stirring evenly, add circulating nitrogen to blow dry, and dry in vacuum to remove residual Organic solvents. After being completely dried, phosphate buffered saline (PBS, pH=7.4) containi...

Embodiment 2

[0027] Polyethylene glycol-modified distearoylphosphatidylethanolamine (DSPE-PEG 2000-Mal) and cysteine-modified cell-penetrating peptide TAT (Cys-TAT) were dissolved at a ratio of 1:1.5 in triethylamine-containing chloroform / methanol (v / v = 2:1) solution and incubated at 25°C for 24 hr in the dark. Excess organic solvent was then removed by vacuum drying, and the precipitate was redissolved in chloroform. After filtering the solution with a filter membrane with a pore size of 0.02 μm, the solution was vacuum-dried again to remove the organic solvent, and the obtained precipitate was stored at -20°C until use.

[0028] Weigh 11.5 mg of soybean lecithin (SPC), 0.96 mg of cholesterol, and 1.15 mg of DSPE-PEG 2000-TAT, dissolve the above materials in chloroform, and after stirring evenly, add circulating nitrogen to blow dry, and dry in vacuum to remove residual Organic solvents. After being completely dried, phosphate buffered saline solution (PBS, pH=7.4) containing 0.1 M pac...

Embodiment 3

[0031] First, gold species were synthesized by adding 25 μL of chloroauric acid (HO 4 , 12 mM), 60 μL sodium borohydride (NaBH 4 , 10 mM), shake quickly and incubate at 25°C for 2hr. For gold nanorod growth, mix 950 μL CTAB (0.1 M), 40 μL HAuCl 4 (12 mM), 6 μL silver nitrate (Ag NO 3 , 0.01 M), 6.4 μL of ascorbic acid (AA, 0.1 M) growth solution was shaken and mixed quickly, so that the color of the solution became colorless. After that, 2 μL of gold species was added to the growth solution, and after rapid shaking for 10 s, the incubation was carried out at 25°C for at least 2 hr. The size of the obtained rod-shaped gold nanoparticles (AuNRs) was about 60 nm×25 nm.

[0032] The obtained gold nanorods were centrifuged three times to wash off the excess CTAB on the surface. After the pellet was resuspended, the Au NRs and sulfhydryl-modified DNA were mixed at a ratio of 1:3000, and sodium chloride (Na Cl) and phosphate buffer (PB) were slowly added. Make the final concentra...

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Abstract

The invention relates to a method for carrying drugs or nano particles into specific target cells based on aptamer and cell-penetrating peptide. The method comprises the following steps: preparing cell-penetrating peptide modified by distearoyl phosphoethanolamine-polyethylene glycol, and preparing lipidosome wrapping the drug or the nano particles and the cell-penetrating peptide TAT and aptamer connected onto the lipidosome. According to the method, the lipidosome is used to carry the drugs or nano particles, and the appropriate aptamer and the cell-penetrating peptide are modified on the lipidosome, so that the drug delivery and target treatment can be improved, an effective means is provided for the diagnosis and treatment of diseases, and the application prospect is wide. The lipidosome is used to carry the drugs or nano particles, so that the biological compatibility of the drugs or nano particles is improved, and the uptake rate of tumor cells for the drugs or nano particles is improved. The method is simple and easy in modification, and the used materials are natural, easy to get, easy to degrade in the body and good in biological compatibility.

Description

technical field [0001] The invention relates to a method for using liposomes to carry drugs, and using nucleic acid aptamers and cell-piercing peptides to improve tumor cell targeting and drug uptake rate. The invention belongs to the field of nano biomedical materials. Background technique [0002] The cell membrane evolved by the organism can effectively prevent foreign substances from entering the cell and ensure the effective functioning of various biological macromolecules in the cell. However, it is difficult for exogenous drugs to directly reach the lesion through the cell membrane. To a certain extent, the cell membrane becomes a natural barrier to the treatment of tumor cells. In order to enhance the therapeutic effect of drugs, a series of methods are used to enhance the permeability of cell membranes, such as cell penetrating peptides, fatty acids, liposomes, polymers or surfactants. Among them, cell-penetrating peptides can carry small molecule drugs, nucleic a...

Claims

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Application Information

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IPC IPC(8): A61K47/64A61K47/54A61K31/704A61K31/337A61K33/24A61P35/00
CPCA61K31/337A61K31/704A61K33/24
Inventor 何丹农徐艳王萍金彩虹
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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