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Combined tumor targeted treatment system based on photodynamic therapy and chemotherapy

A technology of photodynamic therapy and chemotherapy, applied in antitumor drugs, drug combinations, medical preparations containing active ingredients, etc., can solve the problems of low encapsulation efficiency and drug loading, limited wide application, poor stability, etc. Achieve obvious tumor targeting effect in vivo, improve targeting efficiency and powerful killing effect

Inactive Publication Date: 2017-07-07
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The photosensitizer pyropheophorbide-a (PPA) has excellent photodynamic properties, but its wide application is limited due to its poor water solubility; degraded polylactic acid (PLA) nanoparticles, but the encapsulation efficiency and drug loading of co-encapsulation are low, and the stability is poor

Method used

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  • Combined tumor targeted treatment system based on photodynamic therapy and chemotherapy
  • Combined tumor targeted treatment system based on photodynamic therapy and chemotherapy
  • Combined tumor targeted treatment system based on photodynamic therapy and chemotherapy

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1: Preparation and Characterization of Nano Delivery System

[0036] The nano drug delivery system was prepared by emulsified solvent evaporation method; 22.5 mg of PPA-PLA-PEG-PLA-PPA and 2.5 mg of Mal-PEG-PLA were dissolved in 1 ml of dichloromethane solution, and PTX was added to make the concentration 0.5 mg / ml, then add 2 mL of 1% sodium cholate solution; ultrasonicate for 2.4 min in an ice-water bath (interval time 2 s, power 240 W), and 8 ml of 0.5% sodium cholate solution disperse for 5 min. After the dichloromethane was removed by rotary evaporation, centrifuge at 14500 rpm for 1 h at 4°C to obtain ordinary double drug-loaded nanoparticles (PPA NP-PTX); the preparation of drug delivery system with targeting function is based on ordinary nanoparticles On the surface, the sulfhydryl group (-SH) in the polypeptide is covalently linked to the maleimide group (-Mal) on the surface of the nanoparticle, and reacted in the dark for 6 h. The prepared nanopartic...

Embodiment 2

[0038] Example 2: Investigation of the uptake of delivery system by HCT-15 cells and HUVEC cells

[0039] In order to verify that the nano-delivery system has a dual targeting effect on tumor cells and neovascular endothelial cells; its uptake on human umbilical vein endothelial cells (HUVEC) and PTX-resistant human colorectal cancer cells (HCT-15) were investigated respectively Behavior, the implementation steps are: divide the two kinds of cells into 1×10 4 The density of cells / ml was inoculated on the confocal dish, and after 24 hours of incubation, 1 ml of fresh culture medium containing PPA NP-PTX or F3-PPA NP-PTX with different concentrations ranging from 50 μg / ml to 400 μg / ml was added, and Incubate for 1 h under standard conditions. Then they were washed three times with phosphate-buffered saline (pH 7.4, PBS), fixed with 4% paraformaldehyde, and stained with Hoechst 33258. After 15 min, the qualitative results of cell uptake were observed under a confocal microscope...

Embodiment 3

[0041] Example 3: In vitro cytotoxicity investigation of the nano-delivery system

[0042] HUVEC cells or HCT-15 cells in the logarithmic growth phase were seeded on a 96-well plate at a density of 5,000 per well. After 24 h, the culture medium in the well plate was removed, and the PTX and PPA were both 100 ng / Add 100 μl fetal bovine serum-free culture solution containing NP-PTX, PPA NP, PPA NP-PTX and F3-PPA NP-PTX to the concentration of ml, incubate for 4 h under the condition of 4% CO2 and 37 ℃, then take out , with light sources of different intensities (0.005 J / cm 2 - 3.0 J / cm 2 ) to irradiate the cells, and the cells not irradiated by the light source were used as the control. After continuing to culture for 24 h, 10 μl of CCK-8 solution was added. Finally, the absorbance was detected with a microplate reader and the survival rate of cells in each group was calculated. The results showed that with the increase of light intensity, the cell survival rate in the NP-P...

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Abstract

The invention belongs to the field of medicinal preparations, and relates to a targeted nanometer delivery release system aiming at a multidrug resistant tumor and a preparation method of the targeted nanometer delivery release system. According to the system and the preparation method, the photosensitizer, namely, pyrophaeophorbide-a (PPA) is bonded to two ends of the copolymer, namely, hydroxylated-polylactic acid-polyglycolic acid (HO-PLA-PEG-PLA-OH) through a chemical bonding method, thus the product, namely, the photosensitizer-embedded polylactic acid-polyglycolic acid polymer (PPA-PLA-PEG-PLA-PPA) is obtained, then the chemotherapy medicine, namely, paclitaxel is encapsulated physically by adopting an emulsified solvent evaporation method, thus the binary-drug-loading nanometer delivery release system (PPA NP-PTX) is prepared, then F3 peptide with the specific targeting function and the cell penetrating function is modified on the surface of the system, and thus the photodynamic therapy and chemotherapy combined drug delivery nanometer delivery release system with the binary targeting property is prepared. The experiment proves that the delivery system has the strong lethal effect for drug resistant tumor cells, has the obvious in-vivo tumor targeting effect, and has the obvious treatment effect for the mice bearing the multidrug resistant tumor, and thus the combined drug delivery nanometer system has the clinical application prospect.

Description

technical field [0001] The invention belongs to the field of pharmaceutical preparations, and relates to a targeted nano-delivery system for multi-drug resistant tumors and its construction method and application, in particular to a combined tumor targeting treatment system based on photodynamic therapy and chemotherapy and its build method. In the present invention, the tri-block copolymer is used as the carrier material, and the photosensitizer is embedded on the nanomaterial by chemical bonding to prepare nanoparticles, and then the chemotherapeutic drugs are loaded, and the surface of the nanoparticles is modified to have tumor cells and neoplastic tumors. The F3 peptide with blood vessel-specific targeting and cell-penetrating effects constructs a targeted dual-drug delivery system to achieve targeted therapy combining photodynamic and chemotherapy for tumor sites. Background technique [0002] Practice has shown that multidrug resistance has seriously restricted the e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61K47/59A61K31/337A61P35/00
CPCA61K41/0071A61K31/337A61K2300/00
Inventor 陈钧冯兴业姜頔康婷朱倩倩冯静娴蒋天泽
Owner FUDAN UNIV
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