Tumor targeted therapeutic drug carrier as well as preparation method and application thereof

A treatment drug and tumor-targeting technology, applied in biochemical equipment and methods, anti-tumor drugs, and other methods of inserting foreign genetic materials, etc., can solve problems such as difficult control, random carrier loading, and poor therapeutic effect. Achieve large packaging volume, good blood stability, and good tumor treatment targeting effect

Active Publication Date: 2011-07-27
CHENGDU NUOEN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no method to control the maximum loading of the carrier, which makes the loading of the carrier more random and difficult to control, which leads to poor therapeutic effect

Method used

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  • Tumor targeted therapeutic drug carrier as well as preparation method and application thereof
  • Tumor targeted therapeutic drug carrier as well as preparation method and application thereof
  • Tumor targeted therapeutic drug carrier as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: The transfectivity experiment of DOTAP and lecithin in serum-free medium to H1299 cells in different ratios image 3 As shown, the transfection ability of DOTAP decreased with the increase of lecithin addition. 1 μg / μl of pFCB-eGFP was mixed with equal volumes of DL liposomes at different Dotap:Lecithin molar ratios. H1299 cells were inoculated into 12-well culture plates and treated with Opti-MEM culture medium containing 2.5 μl DL:DNA complex for 4 hours for transfection, and then replaced Opti-MEM with culture medium RPMI containing 10% fetal bovine serum, and culture plates were placed in The incubator was incubated for another 48 hours, observed and photographed with a fluorescence microscope at a magnification of 200, and obtained image 3 the result of.

Embodiment 2

[0066] Example 2: Different transfection efficiencies of complexes formed by DOTAP and lecithin and DNA under high or low serum concentration conditions

[0067] Such as Figure 4 As indicated, H1299 cells were seeded into 12-well culture plates at 37°C, 5% CO 2 Incubate overnight. When the cells grow to 30% saturation, the cells are treated with 1ml culture medium containing 1μl, 2.5μl or 5μl DL:pFCB-eGFP, the culture medium is RPMI-1640 containing 10% fetal bovine serum (FBS) or 1ml 100% FBS. The treated cells were gently shaken by hand every 30 minutes. After 4 hours, the transfection culture solution was replaced with 1ml RPMI-1640+10% FBS culture solution, and after 48 hours of cultivation, the photos were observed with a fluorescence microscope at a magnification of 200. 8mM liposomes with different Dotap: lecithin molar ratios and the same A volume of 1 μg / μl pFCB-eGFP plasmid DNA was bound. The photographs shown here are for Dotap:lecithin molar ratios of 20:8 to ...

Embodiment 3

[0068] Example 3: The relationship between the transfection efficiency and the particle size of the complex formed by the carrier formed by DOTAP and lecithin with a molar ratio of 20:9 and DNA

[0069] Such as Figure 5 As shown, the transfection efficiency of the complex is closely related to the particle size of the complex. A DNA mixture containing 0.2 μg / μl of pFCB-eGFP and 1 μg / μl of pFCB-Luc was mixed with an equal volume of DL [20:9]. The complexes were then filtered through filters with a pore size of 1 μm and 0.45 μm., labeled 1 μm and 0.45 μm. A DNA mixture containing 0.2 μg / μl pFCB-eGFP and 0.6 μg / μl pFCB-Luc was mixed with an equal volume of DL [20:9]. The complexes were then filtered through filters with a pore size of 0.2 μm and 0.1 μm., labeled 0.2 μm and 0.1 μm. H1299 cells were inoculated into 12-well culture plates, and the transfection operation was performed when the cells reached 30% saturation within 24 hours. Transfections were performed with 2 μl a...

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Abstract

The invention discloses a drug carrier for general administration through intravenous injection, a preparation method and technology thereof applied to tumor gene therapy, belonging to the field of tumor targeted therapy. The carrier provided by the invention is a novel liposome formed by DOTAP or other analogues and lecithin or derivatives thereof according to a mole ratio of 20: (7-13), can form a stable composite with bioactive substances, and can directionally and efficiently carry the bioactive substances out of the human body to culture and carry into target cells in the human body. Thecomposite provided by the invention has larger carrying capacity; the particle size is greatly decreased, namely, the particle size is optimized to 200nm and below; high transfection efficiency is kept under the condition of high serum concentration; when the carrier package provided by the invention carries cancer suppressor gene or cell suicide gene DNA and the like, the formed composite can betargeted and carried out of the human body to culture, carried away from the human body and carried into the tumor cells in the human body so as to achieve the purpose of carrying out gene therapy onthe tumor.

Description

technical field [0001] The invention relates to a tumor gene therapy technology, in particular to a carrier of a human tumor-targeted gene therapy drug capable of systemic administration, its preparation method and application. Background technique [0002] At present, in the treatment of tumors, most biologically active substances, that is, tumor therapeutic drugs, lack the ability to send themselves into cells. Therefore, direct administration has disadvantages such as low bioavailability and high cytotoxicity. In order to overcome these disadvantages, a large number of carriers have been studied. With the understanding of the molecular mechanism of many diseases at the gene level, gene therapy has become the most promising way to treat many diseases, especially malignant tumors, that is, to use various carriers to encapsulate biologically active substances such as nucleic acids and carry them to drug targets for transduction and treatment. expression for tumor-targeted t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/24A61K47/18A61K48/00A61P35/00
CPCA61K31/713A61K47/186A61K47/18A61K9/1271A61K31/7088A61K48/00C12N2810/10C12N15/88A61K45/06A61K9/1272A61K9/0073A61K47/24A61K47/00A61K9/0019A61P35/00
Inventor 肖虹
Owner CHENGDU NUOEN BIOLOGICAL TECH
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