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Complexes for target killing tumor cell, preparation and application thereof

A technology for tumor cells and complexes, which is applied in the field of preparing and targeting complexes for killing tumor cells. It can solve the problems of high toxicity and limited use of ricin, and achieve low toxicity, good water phase dispersion ability, and good biological phase. capacitive effect

Inactive Publication Date: 2012-01-18
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the combination of RTA and RTB, the natural Ricin is quite toxic, as long as 1 mg of ricin is enough to poison an adult, as long as a few ricin molecules enter the cytoplasm, it is enough to kill the cell, such a high Toxicity limits people's use of ricin

Method used

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  • Complexes for target killing tumor cell, preparation and application thereof
  • Complexes for target killing tumor cell, preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 Preparation of MWNT-RTA complex

[0032] (1) Purification of MWNT

[0033] Put 1g MWNT in a round bottom flask, add 16ml concentrated HNO 3 , refluxed in an oil bath at 140° C. for 4 hours, centrifuged, discarded the supernatant, repeated washing and centrifugation until the supernatant pH ~ 6, and filtered with a microporous membrane with a pore size of 0.45 μm to obtain a black solid.

[0034] (2) Preparation of truncated MWNT

[0035] The purified MWNTs were further truncated by ultrasonication at 40°C for 8 hours in concentrated sulfuric acid and concentrated nitric acid with a volume ratio of 3:1, so that a large number of carboxyl groups appeared at both ends of the tube and part of the tube wall. The cut-off carbon tube dispersion is suction-filtered with a microporous membrane of 0.45 μm in diameter, washed with a large amount of distilled water until the filtrate is close to neutrality, and then the obtained filtrate is centrifuged at 9000 rpm fo...

Embodiment 2

[0038] Example 2 Broad-spectrum killing effect of MWNT-RTA complex on numerous cell lines

[0039] (1) Cell culture

[0040] The cells used in the present invention are all preserved by the laboratory of Professor Zhong Jiang, School of Life Sciences, Fudan University. Among them, the culture medium used for human liver cell HL7702, human cervical cancer Hela cell, and African green monkey kidney cell COS-7 is 10% fetal RPMI 1640 of bovine serum; human breast cancer cell MCF-7 uses DMEM culture medium containing 10% fetal bovine serum and 1% penicillin-streptomycin. The cells used were cultured in a saturated humidity incubator with a volume fraction of 5% CO2 at 37°C, and cells in the logarithmic growth phase were selected for the experiment.

[0041] (2) Determination of cell death rate

[0042] Add 150 μL of the prepared MWNT-RTA complex to the above cells and continue to culture for 22 hours. The cultured cells were removed from the culture medium, digested with trypsin...

Embodiment 3

[0043] Example 3 Targeted Killing Effect of MWNT-RTA-anti-HER-2 Complex on Human Breast Cancer Cell MDA-MB-453

[0044] Add 250 μL MWNT (0.05 mg / mL), 50 μL RTA (2 μM) and 50 μL anti-HER-2 protein (1 μM) into 150 μL PBS (pH=7.4), stir in ice bath for 2-3 hours, at this time the toxin protein and target Proteins can be adsorbed on the surface of carbon nanotubes due to electrostatic, hydrophobic and other weak interactions.

[0045] The culture medium of human breast cancer cell MDA-MB-453 and Chinese hamster ovary cell CHO is DMEM medium. Add 150 μL of the prepared MWNT-RTA-anti-HER-2 complexes to breast cancer cells and normal cells, and continue to culture for 22 hours. The cultured cells were quickly detected by flow cytometry according to the method in Example 2(2). Figure 4 Column 7 in the column is the death data of two strains of cells caused by MWNT-RTA, in which the ratio of the death rate of cancer cells to normal cells is 1:1, and MWNT-RTA-anti-HER-2 (column 8) ha...

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Abstract

The present invention provides a compound for target killing tumour cell, a method for preparing the same and an appliance, which relates to nano-technology and life science field. The nanometer compound is composed of carbon nanotube less than 50nm -1mum, ricin protein A-chain, anti-HFR-2 targeting protein. The short carbon nanotube combines with ricin protein A-chain and anti-HER-2 targeting protein respectively. The compound has high killing rate by acting on human breast cancer cell and normal cell. The preparing method is simple, and the prepared compound has better biocompatibility and better target for tumour cell, and provides new thinking for clinic research of target killing tumour cell.

Description

technical field [0001] The invention relates to the fields of nanotechnology and life sciences, and in particular provides a compound targeting and killing tumor cells, a preparation method and an application. Background technique [0002] Due to the selective permeability of the cell membrane, many biomolecules, especially proteins such as macromolecules, cannot actively pass through the cell membrane. Therefore, finding new biomacromolecule carriers is an urgent need in the fields of drug delivery, gene and protein therapy. [0003] With its comprehensive excellent properties, carbon nanotubes have shown great application value to people. In recent years, the biomolecular functionalization of carbon nanotubes provides an opportunity for the further application of carbon nanotubes in living biological systems. At present, the exploratory research for the purpose of biological application is increasing rapidly, and has gradually become a new research hotspot. In 2003, Fren...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61K47/04A61P35/00
Inventor 孔继烈翁雪香余绍宁王美艳葛晶
Owner FUDAN UNIV
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