Liver cancer suicide gene therapy medicine based on aminated silicon dioxide nanoparticle-CD/TK (cytosine deaminase-thymidine kinase) fused gene compound

A silicon dioxide and nanoparticle technology, applied in the field of tumor treatment in clinical medicine, can solve the problems of side effects, sequelae, large trauma, etc., and achieve the effect of efficient killing

Inactive Publication Date: 2013-10-23
ZHEJIANG CELLPRO BIOTECH
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, the treatment of liver cancer mainly includes surgery, radiotherapy, chemotherapy, immunotherapy and interventional therapy. Among them, surgery is the preferred m

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  • Liver cancer suicide gene therapy medicine based on aminated silicon dioxide nanoparticle-CD/TK (cytosine deaminase-thymidine kinase) fused gene compound
  • Liver cancer suicide gene therapy medicine based on aminated silicon dioxide nanoparticle-CD/TK (cytosine deaminase-thymidine kinase) fused gene compound
  • Liver cancer suicide gene therapy medicine based on aminated silicon dioxide nanoparticle-CD/TK (cytosine deaminase-thymidine kinase) fused gene compound

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Embodiment Construction

[0026] The preferred embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, so that the protection scope of the present invention can be more clearly defined.

[0027] In the examples, the following abbreviations have the following meanings, and the undefined abbreviations use their generally accepted definitions:

[0028] °C = degrees Celsius; min = minutes; sec = seconds; ng = nanograms; mM = millimoles; mL = milliliters; μL = microliters.

[0029] (1) hTERT promoter cloning

[0030] First extract human peripheral blood DNA.

[0031] Take 10 mL of heparin anticoagulated peripheral blood into a 50 mL centrifuge tube, add 2 to 3 times the volume of 1× red blood cell lysate, mix, and let stand on ice for 30 min until the solution becomes transparent. Centrifuge at 4°C at 3000 rpm for 10 min. Clear liquid. ...

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Abstract

The invention provides a liver cancer suicide gene therapy medicine based on an aminated silicon dioxide nanoparticle-CD/TK (cytosine deaminase-thymidine kinase) fused gene compound. A preparation method for the liver cancer suicide gene therapy medicine comprises the following steps of: (1) amplifying CD genes and TK genes via PCR (polymerase chain reaction), and constructing a fused gene, cloning the fused gene into a conventional plasmid vector, promoting the expression of the fused gene by a human-source telomerase promoter in the vector, and enhancing the transcription of the fused gene by a CMV (cytomegalovirus) enhancer; (2) mixing the constructed plasmids with aminated silicon dioxide nanoparticles, so as to form a stable nano-DNA (deoxyribonucleic acid) compound; and (3) taking the nano-DNA compound for therapy for liver cancer as a gene therapy medicine, wherein the medicine is capable of transfecting liver cancer cells in vivo or in vitro, and Cd/TK is capable of being highly expressed in the liver cancer cells, and capable of converting a pro-medicine 5-Fc to a toxic medicine 5-Fu, thus having obvious toxic and killing effects on the liver cancer cells.

Description

Technical field [0001] The invention relates to the treatment of tumors in the medical clinic, in particular to a liver cancer suicide gene therapy drug based on aminated silica nano particles-CD / TK fusion gene complex. Background technique [0002] At present, the treatment of liver cancer mainly includes surgery, radiotherapy, chemotherapy, immunotherapy and interventional therapy. Surgery is the preferred method, but it usually requires a combination of multiple therapies, but these methods have their limitations, such as incompleteness and trauma. Serious, serious side effects and sequelae. Gene therapy is a treatment method that has been developed in recent years, which is different from traditional therapies. It refers to the introduction of foreign genetic material (DNA) into the patient’s somatic cells through a vector and effective expression, or the modification of tumor cells or immune effector cells in vitro and back These cells are transferred to replace mutant gene...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/48A61P1/16A61P35/00
Inventor 曾桥薛志刚
Owner ZHEJIANG CELLPRO BIOTECH
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