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Nucleic acid aptamer cluster modified nanostructure and preparation method and application thereof

A nucleic acid aptamer and nanostructure technology, applied in the field of biomedicine, can solve the problems of limited hybridization sites, achieve low toxicity and side effects, improve cell targeting effect and internalization efficiency, and achieve large-scale production

Pending Publication Date: 2022-07-22
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the sites on the surface of various nanostructures that can hybridize with nucleic acid aptamers are often limited, and the nucleic acid aptamers currently used are mostly monovalent ligands.

Method used

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  • Nucleic acid aptamer cluster modified nanostructure and preparation method and application thereof
  • Nucleic acid aptamer cluster modified nanostructure and preparation method and application thereof
  • Nucleic acid aptamer cluster modified nanostructure and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] In this example, a nanostructure modified by a nucleic acid aptamer cluster is prepared, and the steps are as follows:

[0099] (1) Preparation of linked strand-modified nucleic acid aptamer clusters

[0100] a) Preparation of DBCO-modified linker strands and aptamer strands

[0101] 100 μL NH 2 The C6-modified linker chain SEQ ID NO: 1 (260 μM concentration) was mixed with 39 μL DBCO-NHS (20 mM concentration) dissolved in N,N-dimethylformamide solution, to which 50 μL acetonitrile and 2 μL triethyl acetate were added amine, after stirring for 12 h, sodium acetate with a final concentration of 0.3M and 75% ethanol were added to the reaction system, shaken and mixed, and then placed at -80°C overnight.

[0102] At 4°C, centrifuge at 13,000 rpm for 30 min to precipitate DNA, remove the supernatant, dissolve the obtained precipitate in water, and use a purification column (3 kDa molecular weight cut-off) at 13,000 rpm for 10 min to separate and concentrate to remove exc...

Embodiment 2

[0128] In this example, a nanostructure modified by a nucleic acid aptamer cluster is prepared. The difference from Example 1 is that the target of the nucleic acid aptamer chain is nucleolin, the nucleic acid sequence is SEQ ID NO: 3, and the remaining conditions and implementation Example 1 is the same.

[0129] SEQ ID NO: 3: ggtggtggtggttgtggtggtggtggtttttt.

Embodiment 3

[0131] In this example, a nanostructure modified by a nucleic acid aptamer cluster is prepared. The difference from Example 1 is that in step (3), the molar ratio of the nucleic acid aptamer cluster and the nanostructure is 5:1, and the other conditions are the same as Example 1 is the same.

[0132] After gel electrophoresis detection, it was shown that Example 2 and Example 3 successfully prepared nanostructures modified by polyvalent nucleic acid aptamer clusters. rules, and has good dispersion. The above results are very similar to the results in Example 1, and are not repeated here for the purpose of saving space.

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Abstract

The invention provides a nanostructure modified by an aptamer cluster as well as a preparation method and application of the nanostructure. The aptamer cluster consists of a branched chain type organic molecule, at least two aptamer chains and one connecting chain, the nanostructure contains a capture chain; the nucleic acid aptamer cluster is connected with the nanostructure through the interaction between the connecting chain and the capturing chain. The invention also provides a preparation method of the nanostructure modified by the nucleic acid aptamer cluster. The nucleic acid aptamer cluster is modified on the nanostructure, so that the effective concentration of the nucleic acid aptamer is improved, the purpose of more accurately delivering drugs to target cells, effectively inhibiting the activity of the target cells and inducing the apoptosis of the target cells is achieved, and good specificity and biological safety are achieved; the preparation method is simple, the production efficiency is high, and popularization and application of related products are promoted.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a nanostructure modified by a nucleic acid aptamer cluster and a preparation method and application thereof. Background technique [0002] For many years, tumor has always been one of the malignant diseases with extremely high mortality rate. At present, the clinical methods used to treat tumors mainly include surgical resection, radiotherapy and chemotherapy. However, surgical resection is mostly only suitable for patients with early stage and physical condition tolerable, and it is easy to affect the human immune system and trigger systemic inflammatory response, leading to tumor metastasis and recurrence. Radiotherapy and chemotherapy often lead to severe systemic side effects due to the lack of tumor cell-specific killing. The ultimate goal of tumor therapy is to effectively eliminate tumor cells while avoiding damage to normal cells. Therefore, there is an...

Claims

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Application Information

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IPC IPC(8): C07H21/04C07H1/00A61K31/711A61K47/54A61P35/00
CPCC07H21/04C07H1/00A61K47/549A61K31/711A61P35/00Y02A50/30
Inventor 丁宝全刘建兵武晓慧
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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