Anti-GOLPH2-lipoplexes liposome carrier as well as preparation and application thereof

A technology of liposomes and cationic liposomes, applied in the field of gene carriers, can solve the problems of reduced vitality and number, influence on treatment effect, weakened recognition ability of normal cells and cancer cells, etc., to achieve amplified killing effect, enhanced killing effect, Avoidance of lethal effects

Inactive Publication Date: 2018-07-27
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the activity and number of the receptors are correspondingly reduced under pathological conditions, which w...

Method used

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  • Anti-GOLPH2-lipoplexes liposome carrier as well as preparation and application thereof
  • Anti-GOLPH2-lipoplexes liposome carrier as well as preparation and application thereof
  • Anti-GOLPH2-lipoplexes liposome carrier as well as preparation and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Preparation of liposome lipoplexes.

[0039] Measure 12mL of 0.9% NaCl solution and 1.3mL of absolute ethanol, add each to a 15mL stoppered centrifuge tube, place in a 60°C water bath for 1-2 hours, and bring the solution to 60°C.

[0040] Accurately weigh 100 mg each of DDAB and DOPE, and put them into a centrifuge tube filled with 1.3 mL of absolute ethanol. After each component was completely dissolved, quickly transfer the mixed solution into a centrifuge tube filled with 12mL 0.9% NaCl solution, and quickly invert and mix well. During the above operation, the temperature of the solution was maintained at 60°C. Filter with a filter membrane with a pore size of 100 nm to obtain ordinary cationic liposomes with a particle size of about 100 nm.

[0041] Ordinary cationic liposomes were ultrafiltered with 10 times the volume of water at 15,000 rpm / min for 35 min, and the volume was adjusted to 13 mL to obtain a lipoplexes solution.

Embodiment 2

[0042] Example 2: Construction of Anti-GOLPH2 antibody-modified liposome Anti-GOLPH2-lipoplexes.

[0043] Take 0.5 mL of liposome lipoplexes solution, add 30 μg DSPE-PEG-Mal, and activate in a water bath at 60° C. for 1 hour to obtain an activated liposome lipoplexes solution.

[0044] According to the molar ratio of Anti-GOLPH2 antibody to 2-IT of 1:2, mix the Anti-GOLPH2 antibody solution with a concentration of 0.647mg / mL and the PBS solution of 2-IT, and react at room temperature for 1h to obtain the activated Anti-GOLPH2 antibody .

[0045] Take 100 µL of activated lipoplexes solution, add it to a sterilized 1.5 mL centrifuge tube, add the above-mentioned activated Anti-GOLPH2 antibody to a final concentration of 8 µg / mL, and react at room temperature for 4 h under nitrogen protection to obtain the complex Anti-GOLPH2. GOLPH2-lipoplexes.

[0046] According to the molar ratio of Glycine and 2-IT of 2:1, Glycine and 2-IT were mixed in PBS solution, left to react at room t...

Embodiment 3

[0047] Example 3: Preparation of Anti-GOLPH2-lipoplexes / pBI-Sur-TK complex.

[0048] Add 2 μg of plasmid pBI-Sur-TK to 4 μL Anti-GOLPH2-lipoplexes, mix well, and react at room temperature for 20 minutes to prepare the Anti-GOLPH2-lipoplexes / pBI-Sur-TK complex.

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Abstract

The invention discloses an anti-GOLPH2-lipoplexes liposome carrier, which is prepared from activated cationic liposome lipoplexes and activated anti-GOLPH2 antibodies through crosslinking by water-soluble carbodiimide. The anti-GOLPH2-lipoplexes liposome carrier belongs to a targeted non-virus liposome carrier. The liposome carrier is used for covering recombinant plasmids pBI-Sur-TK to form a compound; the dual targeted and control treatment on the liver cancer is performed; the targeted enrichment and expression of the treatment gene in liver cancer cells are realized; the killing effect onthe liver cancer cells is enhanced; the injury to the normal liver cells is reduced.

Description

technical field [0001] The present invention relates to a gene carrier with biological target recognition function, in particular to a targeted liposome carrier modified based on Anti-GOLPH2 antibody, and a liposome carrier constructed by loading a specific plasmid Complex, the carrier complex constructed in the present invention can specifically kill liver cancer cells. Background technique [0002] As a malignant tumor, primary liver cancer has always been at the top of the list in terms of morbidity and mortality. The current treatment methods for primary liver cancer mainly include surgical treatment, radiofrequency ablation, liver transplantation, chemical drug treatment, physical radiation therapy and emerging biological gene therapy. The prognosis of surgical treatment is poor, and the immune rejection of liver transplantation is large. Chemotherapy and physical therapy are mostly used for the treatment of advanced liver cancer. [0003] Biological gene therapy is t...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K9/127A61K47/42A61K47/24A61P35/00
Inventor 于保锋袁洋洋张丽娜田九博王萱马培元姚志坚曾思衡刘畅张婵穆秀丽孙公勤解军刘志贞郭睿孟凡秀张琪曹睿张紫燕傅松涛杨丽红王惠珍徐钧刁海鹏陈显久胡晓年王海龙弓韬赵虹张栋
Owner SHANXI MEDICAL UNIV
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