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

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0022] Example 1

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

[0024] Weigh 11.5 mg of soybean phospholipids (SPC), 0.96 mg of cholesterol, and 1.15 mg of DSPE-PEG 2000-TAT, and dissolve the above materials in chloroform. After stirring, add circulating nitrogen to blow dry, and vacuum dry to remove residual Organic solvents. After it is completely dry, add phosphate buffer solution (PBS, pH=7.4) c...

Example Embodiment

[0026] Example 2

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

[0028] Weigh 11.5 mg of soybean phospholipids (SPC), 0.96 mg of cholesterol, and 1.15 mg of DSPE-PEG 2000-TAT, and dissolve the above materials in chloroform. After stirring, add circulating nitrogen to blow dry, and vacuum dry to remove residual Organic solvents. After it is completely dried, add 0.1 M paclitaxel-containing phosphate buffer soluti...

Example Embodiment

[0030] Example 3

[0031] First, synthesize gold species, add 25 μL of chloroauric acid (H AuCl) to 750 μL of cetyltrimethylammonium bromide (0.1 M, CTAB) aqueous solution 4 , 12 mM), 60 μL of sodium borohydride in an ice bath (Na BH 4 , 10 mM), shake quickly and incubate at 25°C for 2hr. For growth of gold nanorods, 950 μL CTAB (0.1 M), 40 μL H AuCl 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 quickly shaken and mixed to make the solution colorless. After adding 2 μL of gold seed to the growth solution, shaking rapidly for 10 s, incubating at 25°C for at least 2 hr, the size of the resulting rod-shaped gold nanoparticles (AuNRs) is about 60 nm×25 nm.

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

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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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