Exosome-encapsulated nano drug-loading system for tumor treatment and preparation thereof

A nano-drug loading and tumor treatment technology, applied in the field of nanomaterials and oncology, can solve the problems of poor targeting and killing effect, reduce the proportion of CSCs, etc., and achieve the effect of a mild preparation method

Active Publication Date: 2018-09-18
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention can especially solve the problem of poor targeting and killing effects of CSCs. After the drug-carrying system enters the blood through intravenous administration, it can efficiently accumulate in tumor tissue,

Method used

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  • Exosome-encapsulated nano drug-loading system for tumor treatment and preparation thereof
  • Exosome-encapsulated nano drug-loading system for tumor treatment and preparation thereof
  • Exosome-encapsulated nano drug-loading system for tumor treatment and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: Preparation and characterization of exosome-encapsulated nano-drug delivery system

[0065] 1. Experimental materials and reagents

[0066] H22 mouse liver cancer cells, doxorubicin (with red fluorescence), boron-doped p silicon wafer (resistance 0.00055-0.0015Ω-cm) were purchased from Virginia Company, USA

[0067] 2. Experimental steps

[0068] (1) Place the silicon wafer in concentrated H 2 SO 4 with H 2 o 2 in the mixed solution (volume ratio v:v=3:1) for 15 minutes, washed with ultrapure water for 3 times, and dried in the air for later use. Install the polished side of the silicon wafer in an electrochemical corrosion device, add HF / ethanol solution with a volume ratio of 4:1, 165mA / cm 2 After continuous corrosion for 300s at a certain current intensity, a dark red film appeared on the surface of the silicon wafer, washed with absolute ethanol for 3 times, and continued to add 3.3% HF / ethanol solution (mass ratio), 4.5mA / cm 2 Under the current in...

Embodiment 2

[0075] Example 2: Identification of surface membrane structure of exosome-encapsulated nano-drug delivery system

[0076] 1. Experimental materials and reagents

[0077] FITC-CD63 antibody, CD63 antibody, TSG101 antibody, Calnexin antibody

[0078] 2. Experimental steps

[0079] (1) Take a certain amount of E-PSiNPs, add 500 μL 5% BSA solution and incubate for 30 min, continue to add 10 μL FITC-CD63 antibody, incubate overnight at 4°C with shaking, centrifuge at 20,000g for 30 min, discard the supernatant, and wash 3 times with PBS. Take 20 μL of the E-PSiNPs solution incubated with FITC-CD63 and drop it on the confocal dish. After fully spreading, use the FV1000 confocal microscope to observe the co-localization of the green fluorescence of the FITC-CD63 antibody and the red fluorescence of PSiNPs. The specific parameters are: PSiNPs: Ex=488nm, Em=680nm; FITC: Ex=488nm, Em=520nm.

[0080] (2) Take E-PSiNPs, exosomes and corresponding cells with the same protein amount, and...

Embodiment 3

[0082] Example 3: Uptake behavior of exosome-encapsulated nano-drug delivery system in tumor cells and their CSCs in vitro

[0083] 1. Experimental materials and reagents

[0084] H22 mouse liver cancer cells, doxorubicin, 3D soft fibrin glue, 3D soft fibrin glue (1) In vitro tumor cell uptake behavior

[0085] 2×10 5 H22 was cultured in suspension in the cell culture plate. After 24 hours, the medium was removed, and 1 mL of serum-free DOX, DOX@PSiNPs or DOX@E-PSiNPs with DOX concentrations of 0.5, 1, and 2 μg / mL were added to the cell culture plate, respectively. Medium, 37°C, 5% CO 2 This normal cell culture condition was incubated for 4 hours, the cells were collected, washed 3 times with PBS, and centrifuged at 250g for 10 minutes. Add 500 μL of pre-cooled PBS to the cell pellet to resuspend the cells, pass through a 200-mesh sieve, and detect intracellular DOX fluorescence with a FC500 flow cytometer. Specific detection parameters: Ex=488nm, the emission light is det...

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Abstract

The invention discloses an exosome-encapsulated nano drug-loading system for tumor treatment, and preparation thereof. The system is obtained by utilizing cell endocytosis of a drug-loading nanometermaterial and then exocytosis. The drug-loading nanometer material is loaded with an antitumor medicine including at least one of a chemotherapeutic, a medicine used for immunotherapy and a medicine used for modifying a tumor microenvironment. The composition, structure and the like of the key outer component biofilm encapsulating the nano drug-loading system are improved, and compared with the prior art, the exosome-encapsulated nano drug-loading system provides a novel route for biofilm-based biological processed nanoparticles. By utilization of the exosome-encapsulated nano drug-loading system, the composition and the structure of the exosome can be maintained greatly, and the obtained exosome-encapsulated nano drug-loading system has good stability and tumor targeting performance duringblood circulation. .

Description

technical field [0001] The invention belongs to the field of nanomaterials and oncology, and more specifically relates to a nano drug-carrying system wrapped in exosomes for tumor treatment and its preparation, especially a method for improving the tumor accumulation and deep penetration of anti-tumor drugs 1. Preparation and application of nano-drug delivery system for targeting and killing tumor stem cells. Background technique [0002] Tumor recurrence and metastasis are the most important factors that cause the low survival rate of cancer patients. Studies have shown that tumor recurrence and metastasis are closely related to cancer stem cells (CSCs). The drug resistance of CSCs is one of the main reasons for the failure of tumor therapy, mainly including the following aspects: (1) The high expression of ABC transporter in CSCs is the primary factor for their drug resistance; (2) CSCs are in a quiescent state in G0 phase It is very important to maintain drug resistance...

Claims

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Application Information

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IPC IPC(8): A61K47/46A61K47/04A61K31/704A61P35/00A61P35/02A61P35/04
CPCA61K31/704A61K47/02A61K47/46A61P35/00A61P35/02A61P35/04
Inventor 甘璐雍土莹杨祥良张晓琼别娜娜
Owner HUAZHONG UNIV OF SCI & TECH
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