Nano-compound and its preparation method and application

A nano-composite, gold nano-cages technology, applied in the field of biotechnology and biomedical materials, to achieve the effects of avoiding drug poisoning, inhibiting growth, and protecting drug toxicity damage

Active Publication Date: 2018-07-10
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, after modifying nanomaterials with various tumor targeting strategies, 50% or more of the drug is still enriched in the liver, [Yang Wang, In vivo dual-targeted chemotherapy of drug resistant cancer by rationally designed nanocarrier, Biomaterials ,2016,75,71-81], while chemotherapy drugs enriched in the liver can still produce significant hepatotoxicity

Method used

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  • Nano-compound and its preparation method and application
  • Nano-compound and its preparation method and application
  • Nano-compound and its preparation method and application

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Experimental program
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Effect test

Embodiment 1

[0054] 1. Construction of Au-ODN

[0055] The oligonucleotide (ODN) sequence designed and synthesized in the present invention was purchased from Shanghai Jierui Bioengineering Co., Ltd. The 3' end of one chain of ODN was modified with a sulfhydryl group, and the sequence was as follows: 5'-GCAGCTCGAGCGCTGCGCACGCGT-SH-3'; its complementary sequence was: 3'-CGTCGAGCTCGCGACGCGTGCGCA-5'.

[0056] Gold nanocages (size, 50-100nm) purchased on the market were used to disperse them in phosphate buffer (pH7.0 or so) at a concentration of 0.1-0.5 mg / mL to prepare a gold nanocage solution;

[0057] Add the synthesized ODN to the gold nanocage solution according to the molar ratio of gold and double-stranded oligonucleotide (ODN) at 1:200 to 1:500. Under the action of sulfhydryl, ODN can effectively coordinate with the gold nanocage and bind to the nanocage; after adding the ODN, add an appropriate amount of NaCl solution to the solution to make the final concentration 0.1M; then contin...

Embodiment 2

[0060] 1. Drug capture by nanocomposite Au-ODN

[0061] The chemotherapy drug doxorubicin hydrochloride (DOX) was purchased from Aladdin Company.

[0062] Prepare a solution containing DOX in phosphate buffer solution, so that the concentration of DOX is about 1μM, and then prepare different concentrations of Au-ODN (0.4~3.8nM), add DOX to the Au-ODN solution, react for 30min, and invert for 3~ Mix 5 times.

[0063] The invention designs and synthesizes Au-ODN, which can specifically capture the chemotherapeutic drug doxorubicin (DOX) in the solution. After DOX is captured by Au-ODN, the fluorescence will be weakened or quenched, so it can be accurately detected by the fluorescence quantitative method. The ability of Au-OND to capture DOX. The result is as image 3 as shown in a.

[0064] 2. Stability of Au-ODN in DNase I

[0065] Au-ODN is resuspended in phosphate buffer solution at a concentration of 1-3nM, and then DOX is added to the solution at a final concentration ...

Embodiment 3

[0067] Flow cytometry and laser confocal microscopy were used to detect the entry of Au-ODN into normal liver cells, and CCK8 method was used to detect the protective effect of Au-DON on the viability of normal liver cells under chemotherapy drug treatment. The specific operation is as follows:

[0068] 1. Detection by flow cytometry and confocal laser microscopy

[0069] (1) Normal liver cells QSG-7701 and L02 were mixed with 1×10 4 The density of each well was inoculated in 12-well plates respectively. After 24 hours of culture, Au-ODN resuspended in culture medium was added to the cell culture plate at a concentration of 5-20 μM. The ODN was labeled with fluorescent dye FAM, and the culture was continued for 0 ~4h, and real-time monitoring of Au-ODN entering liver cells by flow cytometry, the results are as follows Figure 4 shown.

[0070] At the end of the culture, the culture medium was aspirated, and the cells were washed 2-3 times with phosphate buffer to remove the...

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Abstract

The invention discloses a nano-compound and its preparation method and application in preparation of a chemotherapeutic drug cytoprotective agent and belongs to the technical field of biotechnology and biomedical materials. The nano-compound comprises a GC base pair-rich oligonucleotide and a gold nanocage. The end 3' of one chain in oligonucleotide is connected to the gold nanocage through a thiol group. The content of the GC base pairs in the oligonucleotide is greater than 50%. The nano-compound Au-ODN can enter cytoplasm through endocytosis, can capture the chemotherapeutic drug entering the cell, and can inhibit the effect of the drug entering the nucleus. 70-80% of the nano-compound can target liver tissue, and only about 5% of the nano-compound can enter cancer tissue. Therefore, the nano-compound Au-ODN can be used for preparing a chemotherapeutic drug cytoprotective agent for preventing liver cells from being damaged by drugs and has no influence on the anti-tumor effect of the chemotherapy drugs.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and biomedical materials, and in particular relates to a nanometer material, a preparation method thereof, and an application thereof in preparing a chemotherapeutic drug cytoprotectant. Background technique [0002] Cancer is one of the diseases with the highest morbidity and mortality in the world. At present, the methods of cancer treatment recognized by the international medical community are still traditional chemotherapy and radiotherapy. However, the use of chemotherapy drugs will cause serious damage to the body, such as liver and kidney failure, hair loss, and emaciation. [0003] In recent years, with the deepening of scientific research, new drugs have continuously entered the market. The anticancer drug (Nivolumab) launched in 2015 is a small molecule drug designed based on the programmed cell death protein PD-1, and has achieved remarkable results in clinical trials. There are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/69A61K47/54A61P35/00A61K31/704
CPCA61K31/704
Inventor 赵瑞波杨新燕刘雪瑶唐睿康
Owner ZHEJIANG UNIV
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