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Intelligent nanoparticle capable of specifically accelerating tumor cell apoptosis and monitoring curative effect by itself

A tumor cell apoptosis and self-monitoring technology, applied in the field of smart nanoparticles and their preparation, can solve the problems of accelerated tumor cell apoptosis, slow drug effect process, lack of curative effect monitoring, etc., achieves good tumor targeting, avoids Acid environment and enzymatic degradation, realizing the effect of timely monitoring

Inactive Publication Date: 2016-08-10
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to propose a smart drug that specifically accelerates tumor cell apoptosis and self-monitors curative effect in order to overcome the problems of poor targeting, low curative effect, slow drug effect process, and lack of curative effect monitoring that exist in most of the current anti-tumor drugs. Nanoparticles have good clinical application value and prospects

Method used

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  • Intelligent nanoparticle capable of specifically accelerating tumor cell apoptosis and monitoring curative effect by itself
  • Intelligent nanoparticle capable of specifically accelerating tumor cell apoptosis and monitoring curative effect by itself
  • Intelligent nanoparticle capable of specifically accelerating tumor cell apoptosis and monitoring curative effect by itself

Examples

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Effect test

Embodiment 1

[0039] Example 1: Combining figure 1 , Synthesize smart nanoparticles that specifically accelerate tumor cell apoptosis and self-monitor curative effect

[0040] 1. Dissolve 1.6mg of hydrophobic antineoplastic drugs (CPT, PTX, DOX, CUR, EVO or SIL), 0.2mg of FRET-Pep, 5mg of PLGA-PEG and 4mg of PLGA-PEI in 2mL of DMSO, stir to dissolve completely;

[0041] 2. Dissolve 1mg DSPE-PEG-FA in 10mL ultrapure water;

[0042] 3. Add the mixture in the above step 1 dropwise to the mixture in the above step 2, and stir for 3-5 hours at room temperature in the dark;

[0043] After stirring, DMSO was removed by ultrafiltration (4000g, 15min), washed with ultrapure water and resuspended to obtain smart nanoparticles that specifically accelerate tumor cell apoptosis and self-monitoring of curative effect, and store at 4°C.

Embodiment 2

[0044] Example 2: Characterization of smart nanoparticles that specifically accelerate tumor cell apoptosis and self-monitor curative effect

[0045] The morphology of intelligent nanoparticles that specifically accelerates tumor cell apoptosis and self-monitoring of curative effect is characterized by JEOL JEM-200CX transmission electron microscope (TEM), and the accelerating voltage is 200kV. The sample solution was dropped on the copper grid of the carbon support film, negatively stained with 2.0% (w / v) phosphotungstic acid solution, and observed under the electron microscope; the size and distribution were characterized by Mastersizer 2000 particle size analyzer dynamic light scattering (DLS). Depend on image 3 It can be seen that the nanoparticle has a spherical core-shell structure, and the nanoparticle has good dispersibility, uniform particle size, and an average particle size of 50-90 nm.

Embodiment 3

[0046] Example 3: Fluorescence response of FRET-Pep to caspase-3

[0047] 100 μL of FRET-Pep was added to 900 μL of reaction buffer, pH 7.4, consisting of 50 mM 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 10 mM dithiothreitol (DTT), 100 mM sodium chloride ( NaCl), 1 mM ethylenediaminetetraacetic acid (EDTA), 0.1% w / v 3-[3-(cholamidopropyl)dimethylamino]propanesulfonic acid inner salt (CHAPS), 10% v / v glycerol (glycerol) . Add different concentrations of caspase-3, and react at 37°C for 1h. Depend on Figure 4 It can be seen that as the concentration of caspase-3 increases, the fluorescence of the polypeptide substrate gradually increases, indicating that the polypeptide substrate can specifically respond to caspase-3.

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Abstract

The invention relates to a preparation method of an intelligent nanoparticle capable of specifically accelerating tumor cell apoptosis and monitoring the curative effect by itself. According to the intelligent nanoparticle, poly(D,L-lactide-co-glycolide)-polyethylene glycol, poly(D,L-lactide-co-glycolide)-polyetherimide and distearoyl phosphoethanolamine-polyethylene glycol-folic acid are adopted as a carrier and covered with hydrophobic anticancer drugs and caspase-3 substrate fluorescent peptide through a one-step self-assembling process. The nanoparticlle can specifically recognize tumor cells overexpressed by folate receptors, and enters lysosome through receptor-mediated endocytosis. By means of the proton sponge effect of polyetherimide, a lysosome membrane is through, escape of the drugs from the lysosome is promoted, tumor cell apoptosis is induced, the iconic caspase-3 of cell apoptosis is activated, the substrate fluorescent peptide is cut, fluorescent signals are generated and used for monitoring the curative effect in real time, precise treatment of tumor cells is achieved, and great tumor diagnosis and treatment application prospects are achieved.

Description

1. Technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to an intelligent nano-particle for specifically accelerating tumor cell apoptosis and self-monitoring of curative effect and a preparation method thereof. 2. Background technology [0002] Tumors are a serious threat to human health and life, and the morbidity and mortality are increasing year by year. At present, chemotherapy plays an important role in the clinical treatment of tumors, but there are still several key issues that have not been resolved: most anti-tumor drugs have no targeting, resulting in large toxic and side effects; the targets of most chemotherapy drugs are located in cells , and the drug usually goes through the lysosomal degradation pathway during the process of entering the cell, resulting in a decrease in the efficacy of the drug and a prolonged time to exert its effect; traditional chemotherapy is difficult to reflect the therapeutic effect...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/34A61K49/00A61K45/00A61K31/4745A61P35/00
CPCA61K9/5146A61K9/5123A61K9/5153A61K31/4745A61K45/00A61K49/0043A61K49/0056
Inventor 余伯阳田蒋为罗荧萍皇立卫
Owner CHINA PHARM UNIV
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