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Hepatic carcinoma targeted photo-thermal therapeutic agent as well as preparation method and application thereof

A photothermal therapy agent, a technology for liver cancer, applied in the field of biomedical engineering, can solve the problems of RGO cannot be independently imaged and has no active targeting ability, and achieve the effect of excellent clinical application value.

Active Publication Date: 2016-11-09
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Photothermal therapy of reduced graphene oxide (RGO) has positive curative effect, but RGO cannot be independently imaged and has no active targeting ability

Method used

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  • Hepatic carcinoma targeted photo-thermal therapeutic agent as well as preparation method and application thereof
  • Hepatic carcinoma targeted photo-thermal therapeutic agent as well as preparation method and application thereof
  • Hepatic carcinoma targeted photo-thermal therapeutic agent as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of Biotinylated Nanobubbles

[0043] ① Select DSPC, DSPE-PEG2000, and DSPE-PEG2000-biotin with a mass ratio of 18:1:1 as raw materials, put 10 mg of phospholipid mixture in a 20 ml round bottom flask, and add a mixed solution of chloroform and methanol drop by drop (volume ratio 2: 1) until the lipid mixture is completely dissolved. Place the flask in a rotary evaporator and evaporate to dryness at 40°C under vacuum;

[0044] ②After evaporating to dryness, add 5ml of PBS buffer solution (pH=7.4, 0.15mol / l) to the flask to redissolve, transfer the liquid into a centrifuge tube, seal it, and replace the internal air with C 3 f 8 ;

[0045] ③ Vibrate with an amalgam oscillator for 40 seconds, then place the solution in a water bath at a temperature of 55° C., and process it with an ultrasonic homogenizer (working frequency 20 kHz) for 20 seconds. After the treatment, incubate at the same temperature for 60 minutes, and then continue the ultrasonic...

Embodiment 2

[0047] Example 2 Cy7 fluorescent labeling of biotinylated anti-GPC3 antibody

[0048] ①Biotinylated anti-GPC3 antibody was purchased from ABCAM Company, UK;

[0049] ②Configuration of cross-linking reaction solution: Accurate NaHCO 3 Powder 0.378g, Na 2 CO 3 Put 0.053g of powder and 0.368g of NaCl powder in a 50ml volumetric flask, add sterile double distilled water to make up to 50ml;

[0050] ③Cy7 powder is dissolved in DMSO, the concentration is 1mg / ml, and the whole process is protected from light;

[0051] ④ Dissolve the antibody in the cross-linking reaction solution at a concentration of 2mg / ml, and react according to the ratio of antibody to Cy7 mass ratio of 1mg to 150ug, slowly add Cy7 fluorescent dye to the antibody cross-linking reaction solution, shake gently to mix, Protect from light and react at 4°C for 8 hours;

[0052] ⑤ Add 5mol / L NH dropwise 4 Stop with 25 μL of Cl solution, and continue to react for 8 hours at 4°C in the dark;

[0053] ⑥ The obtaine...

Embodiment 3

[0054] Embodiment 3 Preparation of biotinylated reduced graphene oxide

[0055] ① Weigh 1g of graphite powder, add about 40g of NaCl powder and mix well, and grind the mixture thoroughly with a ceramic mortar until no obvious graphite particles are visible. Add water to dissolve and filter to wash off the NaCl powder in the mixture, and place the finely ground graphite in an oven to dry. Add the dried graphite powder into a 250mL round-bottomed flask, carefully add about 23mL of concentrated sulfuric acid while stirring, and stir at room temperature for 8 hours until the concentrated sulfuric acid and graphite are fully mixed. Place the reaction vessel in an ice-water bath. After the temperature of the mixture in the flask drops below 15°C, slowly add a total of 3 g of potassium permanganate solids one by one. Stir continuously during the addition to ensure that the temperature in the flask remains uniform and stable during the process. After adding the solid potassium perman...

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Abstract

The invention provides a hepatic carcinoma targeted photo-thermal therapeutic agent as well as a preparation method and an application thereof and belongs to the field of biomedical engineering. The hepatic carcinoma targeted photo-thermal therapeutic agent is prepared with the method comprising the following steps: (1) preparing biotinylation nanometer microbubbles; (2) preparing a Cy7 fluorescently-labeled biotinylation anti-GPC3 (glypican3) antibody; (3) preparing biotinylation RGO (reduced graphene oxide); (4) coupling the products prepared in the step (1), step (2) and step (3) through a biotin-avidin system to obtain the hepatic carcinoma targeted photo-thermal therapeutic agent. The hepatic carcinoma targeted photo-thermal therapeutic agent has ultrasonic and fluorescence imaging functions, mediates RGO targeted delivery with an ultrasound-targeted microbubble destruction technology, monitors a photo-thermal therapeutic process in real time, can be used for preparing drugs for hepatic carcinoma therapy and has excellent clinical application value.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a liver cancer targeted photothermal therapy agent and its preparation method and application. Background technique [0002] Hepatocellular carcinoma (HCC) ranks sixth in cancer incidence and third in mortality in the world. Although surgery is the main treatment for liver cancer, only 9% to 29% of patients can be surgically resected. Photothermal therapy (Photothermal therapy, PTT) is a non-invasive thermal ablation therapy technology developed in recent years. Kill tumor cells. Tumor blood vessels are composed of two parts, one part is the inherent blood vessels of the host. During the process of tumor growth, most of the local microvessels of the host are destroyed, and only the arteries and larger veins may remain and become the backbone of the tumor vasculature. Some blood vessels have normal permeability, and the maximum pore diameter of the vessel wall do...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61K47/48A61K49/00A61K49/22A61P35/00A61P1/16
CPCA61K41/0052A61K49/0034A61K49/0058A61K49/223
Inventor 程文
Owner HARBIN MEDICAL UNIVERSITY
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