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Adriamycin nanoparticles entrapped by bacterial outer membrane vesicles and application of adriamycin nanoparticles

A technology of outer membrane vesicles and doxorubicin, which is applied in the field of medicine, can solve the problems of no induced immune effect, limited clinical application, low immunogenicity, etc., and achieve the goal of prolonging drug half-life, good safety, and prolonging half-life in vivo Effect

Inactive Publication Date: 2021-06-04
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] So far, OMVs have been widely used as delivery vehicles for proteins, plasmids, small interfering RNA (siRNA) and other therapeutic drugs; Escherichia coli with low immunogenicity and human epidermal growth factor receptor 2 (HER2 ) specifically bound OMVs, which can kill HER2+ tumors by delivering siRNA targeting spindle kinesin (KSP), and achieve tumor cell-specific therapeutic effects. However, in the above studies, OMVs were only used as drug delivery vehicles. , and did not exert the ability to induce immune effects; bacteria and their related products are generally considered to be pathogenic to a certain extent, and safety issues limit their clinical application; however, researchers still believe that the OMVs are retained to a certain extent Immunogenicity will aid cancer immunotherapy
[0007] According to literature search, so far, there has been no relevant research on the preparation of OMVs loaded with doxorubicin (DOX-OMV nanoparticles) from OMVs of attenuated Klebsiella pneumoniae and the study of its anti-non-small cell lung cancer effect. to report

Method used

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  • Adriamycin nanoparticles entrapped by bacterial outer membrane vesicles and application of adriamycin nanoparticles
  • Adriamycin nanoparticles entrapped by bacterial outer membrane vesicles and application of adriamycin nanoparticles
  • Adriamycin nanoparticles entrapped by bacterial outer membrane vesicles and application of adriamycin nanoparticles

Examples

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Effect test

Embodiment 1

[0048] Example 1: Preparation and Characterization of DOX-OMV Nanoparticles

[0049] The preparation method of OMVs and DOX-OMV nanoparticles is as above-mentioned, transmission electron microscope and zeta potential analyzer observation show, the OMVs average particle diameter of attenuation Klebsiella pneumonia origin is 71.23nm, and particle diameter increases after packing DOX to be 71.23nm 93.09nm, when the mass ratio of DOX and OMVs is 1:45, the LC-MS detection encapsulation efficiency reaches 78% ( figure 1 ).

Embodiment 2

[0050] Example 2: Uptake of drugs in DOX-OMV by non-small cell lung cancer cells

[0051] Non-small cell lung cancer A549 cells were inoculated on confocal plates, cultured overnight, and respectively treated with OMVs, free DOX (20 μg / ml), and DOX-OMV nanoparticles or DOX liposome nanoparticles (DOX-LIPO) of the same amount of drug Treatment for 12h; or DOX-OMV treatment for 0h, 6h, 12h and 24h. After staining with Hoechst33342 dye, the uptake of drugs by cells was observed by laser confocal microscope. figure 2 The results showed that OMVs could significantly enhance the uptake efficiency of doxorubicin by tumor cells.

Embodiment 3

[0052] Example 3: Anti-non-small cell lung cancer effect of DOX-OMV in vitro

[0053] A549 cells (5×10 3 / well) were inoculated in a 96-well plate, cultured overnight, and treated with different concentrations of DOX, DOX-OMV and DOX-LIPO for 24 hours, the cell viability was detected by MTT method and the IC50 value was calculated. image 3 A The results showed that the IC50 values ​​of DOX, DOX-LIPO and DOX-OMV were 35.51, 12.19 and 11.92 μg / ml, respectively. Western-blot detection of expression of apoptosis-related proteins, image 3 The results of B show that DOX-OMV can significantly induce the splicing and activation of apoptosis-related protein caspase 3 and its downstream key protein PARP. The results of flow cytometry also showed that DOX-OMV could significantly induce tumor cell apoptosis ( image 3 C).

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Abstract

The invention belongs to the technical field of medicines, and relates to drug-loaded nanoparticles, in particular to adriamycin nanoparticles entrapped by bacterial outer membrane vesicles and an application of the adriamycin nanoparticles. The invention provides adriamycin nanoparticles DOX-OMV which are entrapped by bacterial outer membrane vesicles and are prepared by adopting attenuated klebsiella pneumoniae sourced bacterial OMVs (outer membrane vesicles) to entrap a tumor chemotherapeutic drug DOX (doxorubicin). The test research on the effect of resisting non-small cell lung cancer shows that the DOX-OMV nanoparticles can play a lung cancer cell targeting role and an anti-tumor immune induction effect at the same time, and the half-life period of the drug is prolonged, so that the non-small cell lung cancer resisting curative effect of the chemotherapeutic drug adriamycin is remarkably enhanced, and the DOX-OMV nanoparticles have good safety. The prepared DOX-OMV nanoparticles can be used for preparing drugs for resisting non-small cell lung cancer.

Description

technical field [0001] The invention belongs to the technical field of medicine and relates to drug-loaded nanoparticles, in particular to a doxorubicin nanoparticle carried by bacterial outer membrane vesicles and its application. Background technique [0002] According to statistics, the incidence of lung cancer accounts for 11.6% of new cancer cases worldwide, and its mortality rate accounts for 18.4% of global cancer deaths; in China, lung cancer is the cancer with the highest incidence and mortality among men and women. Surveys show that the more common type of lung cancer is non-small cell lung cancer (NSCLC), which accounts for 85% of all lung cancers. In clinical practice, according to the different stages of lung cancer, patients can be treated with surgical resection, radiation therapy and chemotherapy. In recent years, immunotherapy and targeted therapy have made significant contributions to improving the therapeutic effect of NSCLC. However, combined chemotherap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/46A61K31/704A61K39/108A61P35/00
CPCA61K9/5176A61K31/704A61K39/0266A61P35/00A61K2039/522A61K2300/00
Inventor 叶丽库德莱迪·库尔班张慧刘嘉扬
Owner FUDAN UNIV
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