Cancer stem cell-targeting carbon nano-tube-salinomycin drug delivery system, preparation method and uses thereof

A carbon nanotube and delivery system technology, applied in drug delivery, drug combination, pharmaceutical formulation, etc., can solve the problems of non-distributed targeting, stem cell function damage, and limited application, so as to prevent cancer recurrence and metastasis, Effects of inhibiting proliferation and inducing apoptosis

Inactive Publication Date: 2016-04-06
INST OF PHARMACOLOGY & TOXICOLOGY ACAD OF MILITARY MEDICAL SCI P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because salinomycin is difficult to enter into the tumor tissue, it has no targeted distribution, and has no poor selectivity for cancer stem cells and normal tissue stem cells. While killing cancer stem cells, it also causes inhibition and damage to normal stem cell functions, resulting in toxic side effect
At the same time, salinomycin has poor water solubility, and the in vivo administration is only dissolved in ethanol and injected intraperitoneally, which greatly limits its application.
There are no reports on tumor stem cell-targeted drug delivery systems

Method used

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  • Cancer stem cell-targeting carbon nano-tube-salinomycin drug delivery system, preparation method and uses thereof
  • Cancer stem cell-targeting carbon nano-tube-salinomycin drug delivery system, preparation method and uses thereof
  • Cancer stem cell-targeting carbon nano-tube-salinomycin drug delivery system, preparation method and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] The preparation of embodiment 1SAL-SWNTs-CHI-HA

[0124] The preparation of SAL-SWNTs-CHI-HA is a relatively straightforward process, as figure 1 shown. Since commercial SWNTs are too long (>20 μm) to enter cells, and impurities such as metal catalysts and amorphous carbon particles have been shown to be cytotoxic and tissue toxic, the modification previously required purification and oxidation of commercial SWNTs. Oxidation introduces active functional groups such as carboxyl groups and hydroxyl groups at both ends of carbon nanotubes and sidewall defects, and shortens the length of carbon nanotubes, laying the foundation for the next step of functionalization of carbon nanotubes. For the preparation of SAL-SWNTs-CHI-HA, firstly, salinomycin was loaded onto the surface of carbon nanotubes based on the non-covalent hydrophobic interaction between hydrophobic SWNTs and salinomycin, and then chitosan was wrapped around SAL- SWNTs surface to improve their water solubil...

Embodiment 2

[0150] Example 2 Sorting, culturing and identification of gastric cancer stem cells

[0151] It has been reported in the literature that CD44+ gastric cancer cells have the characteristics of gastric cancer stem cells. In this study, the cell surface marker CD44 was used to sort gastric cancer stem cells from AGS gastric cancer cell lines.

[0152] Cell culture and passage: The culture medium used for human gastric cancer AGS cells is DMEM / F12 (1:1), containing 10% fetal bovine serum and antibiotics (penicillin 100U / ml and streptomycin 100μg / ml), at 37°C , 5%CO 2 cultured in an incubator. 0.25% trypsin was used for digestion and passaging.

[0153] Sorting and culturing of gastric cancer stem cells: AGS cells in the logarithmic growth phase were digested with 0.25% trypsin, the digested single cells were collected, washed twice with PBS, and the cell concentration was adjusted to 1×10 6 cells / ml, add the antibody anti-CD44-FITC and incubate at 4°C for 30 min, and finally...

Embodiment 3F

[0158] Example 3 Targeting of FITC-SWNTs-CHI-HA to gastric cancer stem cells

[0159] Flow cytometric analysis: 4×10 CD44+ cells 5 Each well was inoculated in a 6-well plate and incubated with FITC-SWNTs-CHI or FITC-SWNTs-CHI-HA (the final concentration of FITC was 5.0 μM) at 37°C for 3 hours after incubation for 24 hours. Washed three times, digested with 0.25% trypsin, blown into cell suspension with PBS, and measured the fluorescence intensity of FITC bound to cells by flow cytometry (emission wavelength is 488nm, detection wavelength is 520nm). The number of cells used for each analysis was not less than 105, and the number of collected cells was 10,000. Data were analyzed using FCSExpressV3 software. In the receptor competitive inhibition experiment, CD44+ cells were pre-incubated with 5 mg / mL excess free HA for 30 min to saturate the CD44 receptors on the surface of CD44+ cells, and then respectively mixed with FITC-SWNTs-CHI or FITC-SWNTs-CHI-HA (FITC The final con...

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Abstract

The present invention relates to a cancer stem cell-targeting carbon nano-tube-salinomycin drug delivery system, which comprises carbon nano-tubes, drug molecules adsorbed to the carbon nano-tubes, a modification material, and a targeting molecule. The present invention further relates to a preparation method and uses of the carbon nano-tube targeting drug delivery system. According to the present invention, the novel strategy is provided for selective targeting and effective clearing of cancer stem cells, and cancer recurrence and metastasis caused by cancer stem cells can be fundamentally prevented.

Description

technical field [0001] The invention relates to the field of cancer stem cell targeted therapy, in particular to a carbon nanotube-salinomycin drug delivery system targeting cancer stem cells, a preparation method and application thereof. Background technique [0002] Tumor is an abnormal lesion formed by the body under the action of various carcinogenic factors, a certain cell in the local tissue loses its normal regulation of its growth at the gene level, resulting in its clonal abnormal proliferation. The biggest difficulty in tumor treatment is drug resistance, recurrence and metastasis. According to the theory of tumor stem cells, the reason for drug resistance, recurrence and metastasis is the existence of tumor stem cells. [0003] Cancer stem cells are a small number of special cancer cells with self-renewal and multi-directional differentiation potential existing in tumor tissues, which are directly related to the occurrence, recurrence and metastasis of tumors. C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/04A61K47/36A61K47/34A61K47/38A61K31/351A61P35/00A61P35/02
CPCA61K9/0092A61K31/05A61K31/12A61K31/155A61K31/26A61K31/365A61K31/7048A61K47/52A61K47/61A61K47/6923A61K47/6925A61K31/351A61P35/00A61P35/02A61K9/5161A61K9/5192
Inventor 姚红娟张英鸽孙岚刘岩
Owner INST OF PHARMACOLOGY & TOXICOLOGY ACAD OF MILITARY MEDICAL SCI P L A
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