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Liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and preparation method thereof

A technology targeting liposomes and therapeutic effects, applied in preparations for in vivo experiments, drug combinations, anti-tumor drugs, etc., can solve the problems of unmonitored distribution, high toxicity and side effects of chemotherapy, etc., to achieve long-term application, no Toxic and side effects, little damage effect

Inactive Publication Date: 2012-05-02
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problem of high toxicity and side effects of chemotherapy and the inability to monitor its distribution in the body, and to provide a liposome pharmaceutical composition with tumor targeting, in vivo tracing and therapeutic effects and a preparation method

Method used

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  • Liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and preparation method thereof
  • Liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and preparation method thereof
  • Liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Preparation of targeted liposome pharmaceutical composition carrying reporter gene

[0055] (1) DOPC, DOPE, cholesterol, DOPE-PEG2000, DOPE-PEG2000-Mal are mixed according to the original molar ratio of 40:40:40:6:3, and dissolved in methanol;

[0056] (2) utilizing a rotary evaporator to evaporate the solution of step (1) to form a liposome membrane;

[0057] (3) The liposome membrane formed in step (2) was hydrated for one hour with phosphate buffered saline (PBS), so that the total lipid concentration was 3.32 millimoles per liter;

[0058] (4) Ultrasound makes the solution obtained in step (3) become tiny uniform lipid bubbles, and cross the polycarbonate membrane of 100 nanometers to make it a uniform liposome with a particle diameter of about 100 nanometers;

[0059] (5) adding the CD44 antibody according to the molar ratio of the anti-CD44 antibody to DOPE-PEG2000-Mal as 1:50, and using the sulfhydryl group to combine with the PEG chain on the liposome to obtain...

Embodiment 2

[0064] Preparation of non-targeting liposome pharmaceutical composition carrying reporter gene

[0065] (1) DOPC, DOPE, cholesterol, DOPE-PEG2000, DOPE-PEG2000-Mal are mixed according to the original molar ratio of 40:40:40:6:3, and dissolved in methanol;

[0066] (2) utilizing a rotary evaporator to evaporate its solution to form a liposome membrane;

[0067] (3) The liposome membrane formed was hydrated for one hour with phosphate buffered saline (PBS), so that the total lipid concentration was 3.32 mmol per liter;

[0068] (4) Ultrasound makes it become a tiny uniform lipid bubble, and crosses a polycarbonate membrane of 100 nanometers to make it a uniform non-targeted liposome with a particle size of about 100 nanometers;

[0069] (5) Coupling of the reporter gene and the liposome, the liposome is mixed with the plasmid pCDNA3.1-RL-RFP-tTK constructed in Example 1 at a ratio of 20 micrograms of plasmid per 100 microliters of liposomes, and the lipid The positive charge o...

Embodiment 3

[0071] Efficacy identification and in vivo tracking of targeted liposomes

[0072] An orthotopic model of liver cancer in NOD / SCID combined immunodeficiency mice was established. Digest well-grown HepG2 cell lines marked with firefly luciferase and green fluorescent protein with trypsin, wash once with PBS, and resuspend with RMPI1640 basic medium to make the cell density 1×10 per ml medium. 7 cells. Anesthetized with water and chloral, cut the upper left abdomen of the NOD / SCID combined immunodeficiency mouse, exposed the liver lobe, injected 10 microliters of the above cell suspension into the liver lobe of the mouse, and sutured it. About a week later, the liver was in situ A detectable tumor grows. 100 microliters of the targeted liposome-DNA complex prepared in Example 1 was injected into the tumor-bearing mice through the tail vein, followed by intraperitoneal injection of ganciclovir every day at a dose of 50 mg / kg body weight, once a day .

[0073] In vivo chemilum...

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Abstract

The invention discloses a liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and a preparation method thereof. The medicinal composition is a liposome targeting medicament which is resistant to CD44 antibody coupling, has a molecular imaging function simultaneously, and can be used for monitoring the in-vivo distribution of a medicament in real time in a living body state. In particular, plasmids containing three fusion genes, including renilla luciferase, red fluorescent proteins and suicide gene thymidine kinase, are coupled to CD44 antibody mediation-resistant immunoliposome, the specificity of liposome nanoparticles in a liver cancer in-situ model of an in-vivo targeting NOD / SCID (Non-Obese Diabetic / Severe Combined Immune-Deficiency) mouse is monitored by detecting a renilla luciferase signal with a living body imaging system, and apoptosis of liver cancer cells is induced by applying target thymidine kinase of ganciclovir; and moreover, a targeted liposome can be coated with adriamycin for inducing apoptosis of the liver cancer cells. The liposome medicinal composition provided by the invention does not have any toxic or side effect, has small damage and a good effect, and is suitable to be applied for a long time.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to a new liposome pharmaceutical composition with functions of tumor targeting, in vivo tracing and treatment. Specifically, a plasmid containing three fusion genes of renilla luciferase, red fluorescent protein and suicide gene thymidine kinase is coupled on the immunoliposome mediated by anti-CD44 antibody, and renilla luciferase and red fluorescent protein have Tracer function, to judge the distribution of targeted liposomes in the body; the suicide gene thymidine kinase has a therapeutic effect, and the target thymidine kinase of ganciclovir is used to induce apoptosis of liver cancer stem cells. At the same time, anti-CD44 antibody-mediated immunoliposome-encapsulated doxorubicin can target CD44-positive liver cancer stem cells and induce cell apoptosis. Background technique [0002] At present, tumor has become an important disease that threatens human ...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K49/00A61K47/42A61K31/704A61K48/00A61P35/00
Inventor 李宗金向荣王丽娜
Owner NANKAI UNIV
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