Gold nanostar materials cross-linked with pH-responsive transmembrane small peptides and their applications

A gold nano-transmembrane technology, which is applied in the direction of medical preparations containing active ingredients, wave energy or particle radiation treatment materials, non-active ingredients of polymer compounds, etc., to achieve stable properties, increase cell phagocytosis, and biocompatibility Good results

Active Publication Date: 2017-05-31
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, there is no relevant literature report on the preparation of gold nanostar materials with cross-linked pH-responsive transmembrane small peptides and the application of photothermal therapy

Method used

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  • Gold nanostar materials cross-linked with pH-responsive transmembrane small peptides and their applications
  • Gold nanostar materials cross-linked with pH-responsive transmembrane small peptides and their applications
  • Gold nanostar materials cross-linked with pH-responsive transmembrane small peptides and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031]Example 1: Preparation and identification of pH-responsive gold nanostars with cross-linked small peptides with a particle size of about 60 nm.

[0032] (1) Prepare 0.02mL, 2mM maleimide-polyethylene glycol-modified small peptide pHLIP, and 0.02mL, 2mM amino-polyethylene glycol-sulfhydryl with water, mix the two compounds well, and shake well .

[0033] (2) Add 2.5mL of 0.01M chloroauric acid into 97.5mL of water, heat and stir for 5min to 10min, then add 3mL of 1% trisodium citrate, the color of the solution changes from colorless to dark red, stop heating and stirring, A sodium citrate gold solution was prepared.

[0034] (3) Add chloroauric acid (0.25mL, 0.01M) to 10mL water, mix well, add 0.1mL sodium citrate gold solution prepared in (2), then add 0.05mL 0.1M reducing agent ascorbic acid and 0.04mL 0.01 M silver nitrate, the color of the solution changed from light pink to dark blue, and the gold nanostar material was preliminarily prepared.

[0035] (3) After re...

Embodiment 2

[0040] Example 2: Quantitative analysis of cell phagocytosis under pH 6.4 and pH 7.4 conditions

[0041] (1) Under sterile conditions, configure DMEM medium with pH 6.4, add 4.93mL 7.5% NaHCO to 950mL DMEM aqueous medium successively 3 solution and 44.37 mL of 5.2% NaCl solution. Afterwards, 10 mL of penicillin / streptomycin antibiotic mixture was added, and 100 mL of fresh peptide bovine serum was added to obtain a DMEM medium with a pH of 6.4. The DMEM medium with pH 7.4 can be obtained by adding only 10 mL of penicillin / streptomycin antibiotic mixture and 100 mL of fresh peptide bovine serum without adding additional salt solution.

[0042] (2) Cell culture: The breast cancer cell line MCF-7 was divided and grown in DMEM medium with pH 7.4, and placed at 37°C, 5% CO 2 cultured in a cell culture incubator. When the cells grow to a confluence of 70% to 80%, replace the medium with fresh pH 6.4 and pH 7.4 respectively, and dissolve the prepared uncrosslinked and crosslinked ...

Embodiment 3

[0044] Example 3: Identification of pH-responsive gold nanostars cross-linked with small peptides on cell growth inhibition at pH 6.4.

[0045] MCF-7 cells in the logarithmic growth phase were inoculated in a 96-well culture plate. After the confluence of the cells was about 80%, the concentrations of 0mg / L, 0.5mg / L, 1mg / L, 2mg / L and 4mg were added to each well. / L cross-linked small peptide gold nanostar particles dissolved in DMEM medium with pH 6.4, the final volume is 100 μL, set up five duplicate wells, after incubation for 4 hours, discard the supernatant, wash with PBS 3 times, add 100 μL For fresh medium, use 1W / cm 2 The 808nm near-infrared laser was irradiated for 6 minutes, and the control group was not irradiated with laser. After that, 20 μL of tetramethylazolazolium blue (5 mg / mL) was added to each well, and the culture was continued for 4 hours. Sulfone (200 μL / well) was shaken for 10 minutes, and the A570 optical density value was detected. Tumor cell survival...

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Abstract

The invention discloses a pH crosslinking and transmembrane small peptide responding gold nanostar material.Crosslinking of amino-polyethylene glycol-sulfydryl and pHLIP modified with maleimide-polyethylene glycol is conducted firstly, and then the surface of a gold nanostar synthesized with the optimized wet chemistry method is modified to obtain the gold nanostar material.Transmembrane small peptide pHLIP is folded and inserted into cytomembrane to form stable transmembrane alpha-helix in the tumor pH acid environment and helps crosslinking materials enter cells, and then the gold nanostar phagocytosis amount of cells is increased.Under the irradiation of near-infrared laser light, luminous energy absorbed by the gold nanostar is converted into thermal energy, the temperature of a material enrichment region is increased quickly in a short time, tumor cells are killed by means of the thermal effect, and general toxicity is avoided.In vitro study shows that the gold nanostar material can specifically target breast cancer cells in the acid environment, and inhibit growth of tumor cells remarkably after photo-thermal therapy.Due to the fact that pHLIP can be inserted into cytomembrane in a transmembrane mode in the tumor acid environment, the gold nanostar crosslinked with pHLIP can also be widely applied to photo-thermal therapy of other clinical tumors.

Description

technical field [0001] The invention relates to the preparation of a gold nano-star material, in particular to the preparation and application of a gold nano-star material cross-linked with a pH-responsive membrane-penetrating small peptide pHLIP. Background technique [0002] Tumor treatment research has great medical and social significance, and it is a major and urgent need to improve the health of Chinese residents. Traditional anti-tumor treatment mainly includes chemotherapy or physical therapy, but all of them are invasive injuries. For example, certain chemotherapy drugs such as doxorubicin, cisplatin, and mitomycin have great toxicity and side effects. Drug administration will seriously affect the function of normal cells and destroy immune function; most chemotherapy drugs have a short circulation time in the body, it is difficult to target the tumor site to exert their drug effect, and long-term administration will cause some tumor cells to develop drug resistance...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/42A61K41/00A61P35/00
CPCA61K41/0052A61K47/42
Inventor 卢光明田迎张允雷王守巨滕兆刚
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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