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A targeted antitumor complex based on ricin, a preparing method thereof and applications of the complex

A technology of ricin and complexes, applied in the field of biomedicine, can solve the problems of less transfer to the action site and low selectivity of tumor cells, achieve strong cytotoxicity, less toxic and side effects of normal cells, and improve targeted anti-tumor effect Effect

Inactive Publication Date: 2018-02-16
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in order for RTA to give full play to its anti-tumor effect, it is necessary to solve the two problems of low selectivity for tumor cells and less transport to the site of action after entering the cell.

Method used

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  • A targeted antitumor complex based on ricin, a preparing method thereof and applications of the complex
  • A targeted antitumor complex based on ricin, a preparing method thereof and applications of the complex
  • A targeted antitumor complex based on ricin, a preparing method thereof and applications of the complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1, the acquisition of Golgi-targeted CQDs-RTA complex

[0030] Take 200 μL carbon quantum dots (CQDs) with a concentration of 1.8 mg / mL, 5 μL with a concentration of 1 μM RTA and 100 μL 10×PBS buffer (pH 7.4) and mix them in 1 mL of aqueous solution, react in a shaker at 25 °C for 3 h, and store at 4 °C until 0.36mg / mL CQDs-5nM RTA complex was prepared (other high-concentration complexes were prepared in the same way). In order to verify the successful synthesis of the CQDs-RTA complex, it was verified by agarose gel electrophoresis with a mass fraction of 2%, and the results were as follows: figure 1 shown. It can be observed from the figure that the bands of CQDs-RTA complexes have obvious hysteresis relative to the individual CQDs, indicating that the CQDs-RTA complexes have been successfully synthesized.

Embodiment 2

[0031] Embodiment 2, the construction of liposome / carbon quantum dot-A chain complex (Lip / C-R)

[0032] Dissolve soybean lecithin: cholesterol: DSPE-mPEG 2000 (distearoylphosphatidylethanolamine-polyethylene glycol) in a molar ratio of 70:28:2 (10mg, 2.04mg, 1.05mg) in 1 mL of chloroform, 37 ℃ Vacuum rotary evaporation for 30min to form a film, N 2 Blow for 3 minutes, and dry in a vacuum oven at 25°C for 30 minutes to fully remove the organic solvent, then add 450 μL of ethanol to dissolve the lipid layer and inject the resulting solution into the HEPES buffer containing 0.72 mg / mL CQDs-10 nM RTA complex (pH 7.4, containing 25mM HEPES, 150mM NaCl, 5mM KCl, 1mM MgCl 2 and 1mM CaCl 2 ), magnetic stirring at 25°C for 3h, ultrasonication with a 150W probe for 6min, dialysis in a 300kD dialysis bag at 4°C for 12h, and passing through a 0.22μm filter membrane to obtain the Lip / C-R complex. Characterized by scanning electron microscopy (SEM), the results are as follows figure 2 ...

Embodiment 3

[0033] Example 3, Construction of nucleolin nucleic acid aptamer / liposome carbon quantum dot-A chain complex (Apt / Lip / C-R)

[0034] Take 0.6mL of the Lip / C-R complex obtained above, mix it with 25μL 10μM NCL aptamer, 0.1ml 10×HEPESbuffer (pH 7.4) in 1mL aqueous solution, react overnight at room temperature on a four-dimensional mixer, pass through a 100k ultrafiltration membrane (8000rpm / 10min) to remove unbound nucleic acid aptamers, and store at 4°C to obtain the Apt / Lip / C-R complex.

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Abstract

The invention relates to a targeted antitumor complex based on ricin, a preparing method thereof and applications of the complex. The complex is prepared by compositing carbon quantum dots and a ricinA chain to form a carbon quantum dot-ricin A chain complex, then encapsulating the complex with the lipidosome, and modifying the lipidosome with a nucleolin nucleic acid aptamer. The prepared complex can enter cells more easily, acts on ribosome through a Golgi apparatus path, has higher cytotoxicity, can effectively protect the A chain from being enzymatically decomposed by lysosome, utilizes specificity of the nucleolin nucleic acid aptamer, can be targeted on a tumor cell, reduces toxic and side effects on normal cells, and is of great importance for targeted treatment on tumor cells.

Description

technical field [0001] The invention belongs to the field of biomedicine, relates to a targeted anti-tumor compound based on ricin, and also relates to a preparation method and application of the compound. Background technique [0002] Cancer is the second leading cause of death worldwide, and millions of people die from cancer every year. In order to deal with the tumor crisis, many plant toxins have been developed by researchers into new anti-cancer agents with no or minimal side effects. Ricin, as a natural plant protein toxin extracted from castor bean, has great application potential in inducing tumor cell apoptosis as an anticancer agent. Ricin is composed of chain A (RTA) for toxicity and chain B (RTB) for agglutination. RTA inhibits protein synthesis by attacking specific adenine bases on ribosomes. However, RTA alone cannot cross the membrane smoothly, and can only enter cells to play a role by means of the agglutination of RTB. After the complete Ricin enters th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/69A61K47/54A61K47/52A61K38/16A61P35/00
CPCA61K38/168
Inventor 甄淑君李春宏
Owner SOUTHWEST UNIV
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