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Docetaxel drug-loading nanoparticles, preparation method and application thereof

A docetaxel nano-loading technology, which is applied in the field of docetaxel nano-loaded particles, to achieve the effects of reduced toxicity, low cost, and convenient operation

Active Publication Date: 2015-11-25
SUZHOU NANO HEALTH BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on EGF1-multilineage paclitaxel nano drug-loaded particles has not been reported yet.

Method used

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  • Docetaxel drug-loading nanoparticles, preparation method and application thereof
  • Docetaxel drug-loading nanoparticles, preparation method and application thereof
  • Docetaxel drug-loading nanoparticles, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: The preparation of docetaxel-coated nanoparticles includes the following steps:

[0029] 1) Weigh out 1 mg of docetaxel and 50 mg of maleimide-polyethylene glycol-polylactic acid-hydroxyglycolic acid polymer (Malemide-PEG-PLGA) and dissolve in 1ml of dichloromethane, shake to make it fully dissolved, The organic phase is obtained.

[0030] ) Under the condition of ice bath ultrasound, slowly and uniformly add the organic phase to 20ml of 1% polyvinyl alcohol aqueous solution with a syringe within 90s of continuous ultrasound, and continue to set the ultrasound for 12min (parameter setting, power is 300w, 1s on and 1s off ) Obtain O / W type emulsion.

[0031] ) The cling film is pierced and sealed, and the dichloromethane is slowly volatilized by magnetic stirring for 10 hours, and the nanoparticles are uniformly solidified.

[0032] ) High-speed centrifugation of the nanoparticle suspension (instrument parameter setting: 16000rpm, 25°C, 60min), and the supernata...

Embodiment 2

[0036] Example 2: Preparation of EGF1 protein and sulfhydryl modification

[0037] One: The preparation process of EGF1 protein includes the construction of pET28a-EGF1 plasmid, transformation, protein induced expression and purification and identification.

[0038] ) Cut the EGF1 coding gene and pET28a plasmid with EcoRI and HindⅢ, and then ligate the linear DNA with the same restriction site to obtain pET28a-EGF1 plasmid.

[0039] ) Transform the pET28a-EGF1 plasmid into BL21(DE3) strain, screen positive colonies on an agar plate containing 100ug / ml ampicillin, pick the positive colonies, inoculate them in 5ml LB medium containing 100ug / ml ampicillin, shake 24h;

[0040] 3) Inoculate 1L fresh LB culture medium (containing 100mg / L kana) according to 1:100, and culture for 6h in a shaker at 250rpm;

[0041] 4) Add IPTG to a final concentration of 1mmol / L, induce expression at 23°C at 200rpm for 4h, harvest BL21 bacterial solution; 4°C, 10000g, centrifuge for 10min to collect the ...

Embodiment 3

[0053] Example 3: Modification of EGF1 protein on the surface of nanoparticles coated with docetaxel

[0054] 1) Take 5ml 0.01mol / L PBS, add the freeze-dried docetaxel nano drug-loaded particles, pipette to disperse, mix and transfer to a small beaker;

[0055] 2) Add the thiolated EGF1 protein to the above solution according to the molar ratio of -SH:MAL-PEG-PLGA of 1:1;

[0056] 3) Room temperature, avoid light, charge N 2 Stir overnight with low-speed magnetic force;

[0057] 4) Centrifuge the stirred solution at 14000 rpm at 4°C for 60 min, and remove the supernatant;

[0058] 5) Add 2.0ml0.01mol / LPBS, pipette to disperse and mix;

[0059] 6) Pass the solution obtained in step 5 above through a Sepharose CL-4B column to remove unbound free proteins, and collect the eluate containing DTX-EGF1-NP.

[0060] ) DTX-EGF1-NP is freeze-dried to obtain nano-particle freeze-dried powder.

[0061] ) Characterization of DTX-EGF1-NP: The nanoparticle powder was resuspended in 0.001mol / L ...

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Abstract

The present invention discloses docetaxel drug-loading nanoparticles, a preparation method and application thereof. The docetaxel drug-loading nanoparticles include nanoparticles, docetaxel wrapped in the nanoparticles, and sulfhydrylation EGF1 protein modified to nanoparticle surface through covalent bond. The nanoparticles are formed from a maleimide-polyethylene glycol- polylactic acid-hydroxy acid polymer. The nanoparticles disclosed by the present invention have good biological targeting, can actively target tumor cells or tumor vascular cells overly expressing tissue factor, and effectively improve the drug concentration at the tumor site, and reduce the toxic and side effects of the drug.

Description

technical field [0001] The invention relates to the field of biomaterials and nanotechnology, in particular to a docetaxel nano drug-loaded particle with high encapsulation efficiency and drug loading capacity, high biological safety, and capable of slowly releasing the drug docetaxel, and a preparation method thereof and apply. Background technique [0002] Cancer is the second most serious disease that threatens human health and life after cardiovascular disease worldwide. The incidence of breast cancer ranks second in the whole cancer, and it is the most common malignant tumor among women in my country, and the incidence is increasing year by year. The current treatment methods for breast cancer include surgery, radiotherapy, chemotherapy, endocrine therapy and molecular targeted therapy, but surgical resection is still the main method. Molecular targeted therapy can improve the efficacy of tumor chemotherapy drugs and reduce their toxic and side effects. It is the most...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/16A61K31/337A61K47/34A61K47/42A61P35/00A61K47/64A61K47/69
Inventor 李凤英李超
Owner SUZHOU NANO HEALTH BIOTECH
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