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Double targeting liposome with 4-amino-benzene-alpha-D-mannopyranoside (MAN) and wheat germ agglutinin (WGA) modifiers and preparation method thereof and application

A technology targeting liposomes and liposomes, applied in liposome delivery, medical preparations containing no active ingredients, medical preparations containing active ingredients, etc., can solve the problem of undetected epirubicin, epirubicin, Doxorubicin can not reach the therapeutic effect of glioma and other problems, so as to improve the effect of chemotherapy

Active Publication Date: 2015-11-18
SHIHEZI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In some in vitro research experiments, epirubicin has a strong inhibitory effect on glioma cells, however, in vivo optical imaging observations show that epirubicin is not detected in brain regions protected by the blood-brain barrier
The reason is that due to the existence of the blood-brain barrier, epirubicin cannot reach the brain tissue to exert a therapeutic effect on glioma

Method used

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  • Double targeting liposome with 4-amino-benzene-alpha-D-mannopyranoside (MAN) and wheat germ agglutinin (WGA) modifiers and preparation method thereof and application
  • Double targeting liposome with 4-amino-benzene-alpha-D-mannopyranoside (MAN) and wheat germ agglutinin (WGA) modifiers and preparation method thereof and application
  • Double targeting liposome with 4-amino-benzene-alpha-D-mannopyranoside (MAN) and wheat germ agglutinin (WGA) modifiers and preparation method thereof and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, the preparation of liposome

[0059] 1. Preparation of blank liposomes

[0060] 1. Preparation of blank liposomes

[0061] Egg yolk lecithin, cholesterol, DSPE-PEG 2000 、DSPE-PEG 2000 -COOH and DSPE-PEG 2000 -NH 2 Dissolve in methanol at a molar ratio of 52:43:4:0.5:0.5, remove methanol by rotary evaporation, and form a film in an eggplant-shaped flask by rotary evaporation at 35°C and 40rpm, then add 250mM ammonium sulfate aqueous solution, and then perform ultrasonic treatment (ultrasonic parameters : Every 10s of work, stop for 30s, the whole time is 10min, and the protection temperature is 35°C), then use a liposome extruder to pass through the 400nm polycarbonate membrane 3 times and the 200nm polycarbonate membrane 3 times (the sequence is as follows: 400nm polycarbonate membrane Carbon lipid film → 400nm polycarbonate film → 400nm polycarbonate film → 200nm polycarbonate film → 200nm polycarbonate film → 200nm polycarbonate film), then packed i...

Embodiment 2

[0084] Embodiment 2, the performance measurement of liposome

[0085] 1. Determination of Encapsulation Efficiency

[0086] Liposome A1, liposome A2, liposome B1, liposome C1, and liposome D1 obtained in Example 1 were respectively subjected to the following (1) or (2) operations.

[0087] (1) Liposome A1, liposome A2, liposome B1, liposome C1, and liposome D1 were respectively subjected to the following operations: liposomes were placed in a 10mL volumetric flask, and 10 times the volume of methanol was used to Destroy (obtain demulsification solution), detect the concentration of resveratrol and epirubicin.

[0088] (2) Liposome A1, liposome A2, liposome B1, liposome C1, and liposome D1 were respectively subjected to the following operations: the liposomes were passed through a shephedexG-50 column, buffered with PBS of pH 7.4 solution to remove free resveratrol, then packed in a dialysis bag and dialyzed in PBS buffer at pH 7.4 to remove free epirubicin, and then placed i...

Embodiment 3

[0098] Example 3. Research on the inhibitory effect of drug-loaded liposomes on C6 glioma cells in vitro

[0099] When the added form of medicine is liposome, the added amount of epirubicin hydrochloride is the added amount of liposome multiplied by the encapsulation efficiency.

[0100] 1. Take C6 rat brain glioma cells in the logarithmic growth phase, inoculate 3000 cells / well (190 μL / well) in a 96-well plate, place at 37°C, 5% CO 2 Conditioned for 24h.

[0101]2. After completing step 1, take the 96-well plate, and add 10 μL of drug solutions with serial concentrations prepared by PBS buffer solution of pH 7.4 to each well (the addition forms of the drugs are respectively: epirubicin hydrochloride, liposome A1, liposome A2, liposome B1, liposome C1 or liposome D1, make the concentration of each test group epirubicin hydrochloride be 0.05, 0.1, 0.5, 1, 2.5, 5 or 10 μ M respectively; Use an equal volume of PBS buffer to replace the blank control of the drug solution) at 37°...

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Abstract

The invention discloses a double targeting liposome with 4-amino-benzene-alpha-D-mannopyranoside (MAN) and wheat germ agglutinin (WGA) modifiers and a preparation method thereof and application. The double targeting liposome is composed of a liposome and modifiers on the surface of the liposome. The modifiers on the surface of the liposome are 4-amino-benzene-alpha-D-mannopyranoside and wheat germ agglutinin. The invention further provides a medicine carrying liposome which is obtained by using a targeting liposome to perform entrapment of medicines. The medicines can concretely be epirubicin hydrochloride and / or resveratrol. According to the double targeting liposome, double ligands are adopted to perform modification and entrapment of the two medicines, the double targeting effect is achieved, so that the medicines are targeted to the glioma part when striding the blood-brain barrier and are enriched at the tumor position; meanwhile, the two medicines in entrapment can have synergistic or additive curative effect on the focus position, and the curative effect of chemotherapy is improved. A novel idea is provided for preparing medicines for treating glioma, and the double targeting liposome is of great theoretical significance and clinical significance.

Description

technical field [0001] The invention relates to a double targeting liposome with MAN and WGA modification, its preparation method and application. Background technique [0002] At present, the clinical treatment of glioma is still facing great challenges. Non-invasive diagnosis and treatment has become a new direction in the treatment of glioma, which brings hope for improving the treatment effect and prolonging the survival time of patients. However, most of the therapeutic drugs do not have a good therapeutic effect. The reasons are as follows: (1) Due to the existence of the blood-brain barrier, 98% of small molecule drugs and almost all macromolecule drugs cannot enter the tumor site inside the brain to play a therapeutic role. (2) Therapeutic drugs that can penetrate the blood-brain barrier cannot be well enriched in the tumor site, and most of them spread to the normal brain tissue, causing serious adverse reactions and even death of the patient. Therefore, in order t...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K47/26A61K47/42A61K47/34A61K31/704A61K31/05A61P35/00
Inventor 应雪轩亚茹王亚华唐辉
Owner SHIHEZI UNIVERSITY
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