Genetic drug for targeted treatment of liver cancer and its preparation process

A technology for the treatment of liver cancer and gene medicine, which is applied in the field of medical medicine and gene medicine, and can solve problems such as unsatisfactory curative effect

Inactive Publication Date: 2006-04-26
华中科技大学同济医学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Like other tumors, surgery and chemotherapy are often used in the treatment of liver cancer, but the curative effect is not satisfactory

Method used

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  • Genetic drug for targeted treatment of liver cancer and its preparation process
  • Genetic drug for targeted treatment of liver cancer and its preparation process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Preparation of Asor-PLL linker

[0020] Preparation of Asialomucin (Asor) from Human Plasma: Ion Exchange Chromatography and Ammonium Sulfate Fractionation Technique with DEAE-cellulose as Filler [See Whitehead P.H, Sammons H.G. et al Asimpletechnique for the isolation of orosomucoid from normal and pathological sera.Biochim.Bioiphy.Acta, 1966; 124:209-211], extract human serum mucin (Orosomucoid, OR) from human plasma, and then remove sialic acid in human serum mucin by acid hydrolysis , to produce asialo mucin (Asor).

[0021] Human serum mucinoid (Orosomucoid, OR)

[0022] ↓Acid hydrolysis to remove sialic acid

[0023] Asialomucin (Asor)

[0024] Link asialomucin (Asor) with poly-L-lysine (PLL) with protein cross-linking agent water-soluble carbodiimide (EDC) to prepare asialomucin (Asor)-poly-L-lysine Amino acid (PLL) linker (Asor-PLL), specifically, mix Asor, PLL, and EDC at a mass ratio of 1:1:0.5, dissolve in distilled water, adjust the pH to 7.2-7.4...

Embodiment 2

[0048] The asialyl mucin (Asor)-poly-L-lysine (PLL) linker and the pcDNA3 eukaryotic expression vector containing chicken anemia virus vp3 gene were dissolved in 0.9% NaCl solution respectively, and the two solutions were mixed by mass ratio ( Protein: nucleic acid) is 4: 1 fully mixed after, 37 ℃ places 16 hours, through filter sterilization, the prepared concentration is 200 μ g / ml (with the pcDNA3 eukaryotic expression vector containing chicken anemia virus vp3 gene in 0.9% NaCl solution Concentration calculated by mass) drug injection preparation of the present invention.

Embodiment 3

[0050] Asialomucin (Asor)-poly-L-lysine (PLL) linker and the pcDNA3 eukaryotic expression vector containing chicken anemia virus vp3 gene were dissolved in phosphate buffered saline (which consists of 0.8% NaCl, 0.02% KCl, 0.144% Na 2 HPO 4 , 0.024% KH 2 PO 4 ), the two solutions were fully mixed according to the mass ratio (protein:nucleic acid) of 3:1, placed at 37°C for 16 hours, and sterilized by filtration to obtain a concentration of 200 μg / ml (containing chicken anemia in phosphate buffer saline The concentration calculated by the mass of the pcDNA3 eukaryotic expression vector of the virus vp3 gene) the pharmaceutical injection preparation of the present invention.

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Abstract

The invention relates to a genetic drug for targeted treatment of liver cancer and its preparation process, which is the compound of asialoorosomucoid-poly-L-lysine adaptor and eukaryotic expression vector containing Chicken anemia virus vp3 genes, the eukaryotic expression vector is pcDNA3 or pcDNA3.1, the mass ratio (protein : nucleic acid) of the asialoorosomucoid-poly-L-lysine adaptor and the eukaryotic expression vector containing Chicken anemia virus vp3 genes is 3-6:1.

Description

technical field [0001] The invention relates to medical medicine, in particular to gene medicine, especially gene medicine for treating liver cancer and its preparation method. Background technique [0002] Liver cancer is one of the five common malignant tumors, and its incidence is increasing worldwide, seriously endangering human life and health. Like other tumors, surgery and chemotherapy are often used in the treatment of liver cancer, but the curative effect is not satisfactory. With the gradual perfection of gene molecular biology theory and technology, gene therapy brings hope to the treatment of liver cancer. How to achieve targeted gene therapy? This is the first question gene therapy researchers need to consider. [0003] Direct killing of tumor cells is one of the main strategies of liver cancer gene therapy. Whether the therapeutic gene only targets liver cancer cells without damaging normal liver cells is the primary issue worthy of attention. That is, it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P35/00
Inventor 屈伸
Owner 华中科技大学同济医学院
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