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A kind of ATP responsive nano micelles, preparation method and application

A technology of nanomicelle and nanomicelle solution, applied in the field of bioengineering, can solve the problems of non-viral gene carrier targeting and low loading efficiency, achieve high drug loading and encapsulation rate, enhance stability and smooth surface Effect

Active Publication Date: 2021-02-02
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to provide an ATP-responsive nano-micelle, preparation method and application to solve the technical problem of low efficiency of non-viral gene carrier-specific targeting and loading in the prior art

Method used

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  • A kind of ATP responsive nano micelles, preparation method and application
  • A kind of ATP responsive nano micelles, preparation method and application
  • A kind of ATP responsive nano micelles, preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0048] Embodiment 1 Preparation of polylysine-phenylboronic acid and dopamine-hyaluronic acid

[0049] (1) Preparation of polylysine-phenylboronic acid:

[0050]

[0051] According to the above polymer synthesis route, the specific description is as follows:

[0052] Add 4-(bromomethyl)phenylboronic acid and polylysine into anhydrous dimethyl sulfoxide, stir and react at 80°C for 24 hours, the 4-(bromomethyl)phenylboronic acid and polylysine The molar ratio of lysine is 8:1-10:1 respectively, and finally the product is dialyzed and freeze-dried to complete the preparation of polylysine-phenylboronic acid. H NMR spectrum such as image 3 showed that polylysine-phenylboronic acid (PLL-PBA) was successfully synthesized. Note: PLL-polylysine, PBA-phenylboronic acid.

[0053] (2) Preparation of dopamine-hyaluronic acid:

[0054]

[0055] The synthetic route of dopamine-hyaluronic acid (HA-Cat), wherein HA is hyaluronic acid, Dopamine is dopamine, and EDC is 1-(3-dimethyl...

Embodiment 2

[0058] Example 2 Preparation of HA / Bcl-2 siRNA-PLL-PBA-p-gp siRNA-DOX nanomicelles

[0059] Put 5 mg of PLL-PBA and 2.5 mg of Bcl-2 siRNA and p-gp siRNA into 10 mg of DOX aqueous solution to form a Bcl-2 siRNA-PLL-PBA-p-gp siRNA-DOX nanocomplex loaded with genes and drugs, Then put HA into the above-mentioned nano-micelle solution and stir to form HA / Bcl-2 siRNA-PLL-PBA-p-gp siRNA-DOX nano-micelle. The results of the nanomicelles by transmission electron microscopy are as follows: Figure 4 As shown, the prepared nanoparticles are circular and have good dispersibility, and the particle size is mainly concentrated between 100-200nm, especially the particle size of 150nm. Note: Dox is doxorubicin hydrochloride, nm-nanometer.

Embodiment 3

[0060] Example 3 Cytotoxicity of HA / PLL-PBA

[0061] MCF-7 cells were plated in 96-well plates, the density of cells per well was 1×105, and cultured overnight at 37°C in a cell culture incubator with a volume fraction of CO2 of 5%. Nanomicelles of different concentrations were added to the 96-well plate (the cell group with only the culture medium was the negative control group), and 5 replicate wells were set for each experimental condition. After 24 hours of culture, 20 μL of CCK8 (5 mg / mL) solution was added, and the culture was continued for 2 hours. After that, the culture solution was aspirated, tested at 450 nm in a microplate reader, and the cell survival rate was calculated. Among them, CCK8 is a rapid and highly sensitive kit for cell viability and cytotoxicity detection.

[0062] The experimental results are shown in 5. Compared with the control group, the OD value of the cells incubated with nanoparticles within 24 hours did not change significantly, indicating t...

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Abstract

The invention provides an intracellular ATP-responsive nano-micelle. The intracellular ATP responsive nano-micelle consists of polylysine-phenylboronic acid, dopa-hyaluronic acid and a drug resistancereversion gene. The particle size is 100-200 nanometers, the molecular weight of polylysine is 2000-6000 Daltons, and the polylysine is spherical. The invention also provides a preparation method ofthe ATP-responsive nano-micelle and an application of the ATP-responsive nano-micelle in preparation of drugs for treating cancers and as a drug carrier. The nano-micelle can efficiently load and deliver RNA and anti-cancer drugs, can effectively reverse multi-drug resistance of the anti-cancer drugs and can realize tumor targeting of drug-resistant breast cancer. The preparation method is simpleand convenient and is suitable for large-scale production.

Description

technical field [0001] The invention belongs to the field of bioengineering, and relates to a tumor-targeted drug delivery technology, in particular to an ATP-responsive nano-micelle, a preparation method thereof, and an application in a drug for treating tumors. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, and its incidence is increasing year by year. In developed cities such as Shanghai and Beijing in China, the incidence of breast cancer ranks first among new malignant tumors in women. Breast cancer is one of the solid tumors that are sensitive to chemotherapy. Chemotherapy plays an irreplaceable role in the comprehensive treatment of breast cancer. Although the treatment options for breast cancer have been continuously improved, many patients still experience recurrence and metastasis after standard chemotherapy, which leads to The main reason for this phenomenon is that tumor cells develop multidrug resistance (multid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/69A61K31/704A61K47/54A61K47/60A61K48/00A61P35/00B82Y5/00B82Y30/00B82Y40/00
CPCA61K31/704A61K48/0041A61K47/542A61K47/60A61K47/6907A61P35/00B82Y5/00B82Y30/00B82Y40/00
Inventor 刘培峰刘红梅马丁孔宪明
Owner SHANGHAI INST OF ONCOLOGY
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