Thrombin-loaded DNA (deoxyribonucleic acid) self-assembly nano-structure, as well as preparation method and application thereof

A nanostructure, thrombin technology, which is used in medical preparations with non-active ingredients, medical preparations containing active ingredients, drug combinations, etc., can solve the problems of uncomfortable intravenous injection and short half-life of thrombin, and achieve uniform size, Good stability and good biocompatibility

Active Publication Date: 2014-07-30
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the high procoagulant activity of thrombin itself makes it unsuitab...

Method used

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  • Thrombin-loaded DNA (deoxyribonucleic acid) self-assembly nano-structure, as well as preparation method and application thereof
  • Thrombin-loaded DNA (deoxyribonucleic acid) self-assembly nano-structure, as well as preparation method and application thereof
  • Thrombin-loaded DNA (deoxyribonucleic acid) self-assembly nano-structure, as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Preparation of DNA self-assembled nanostructure loaded with thrombin in the present invention

[0058] Specific steps are as follows:

[0059] (1) Preparation of rectangular DNA origami structure

[0060] The M13 phage genomic DNA backbone and staple strands were mixed, and the final concentrations of the backbone and staple strands were 10 nM and 80 nM, respectively. The mixture was slowly annealed using a gradient PCR instrument. The annealing conditions were as follows: from 65° C. to 25° C., each 1° C. was a gradient, and the residence time of each gradient was 10 minutes. The entire annealing process was about 7 hours.

[0061] After the annealing procedure was completed, the DNA origami structure samples were taken out and centrifuged with a 100kDa spin column (MWCO) to remove excess staple strands and capture strands. The centrifugation conditions are: take 100 μL sample, add 350 μL 1×TAE-Mg buffer solution, centrifuge at 4700 rpm for 3 minutes, the ...

Embodiment 2

[0067] Example 2: Anti-tumor effect of the DNA self-assembled nanostructure loaded with thrombin in the present invention

[0068] Evaluation method of anti-tumor effect:

[0069] will be 2.0×10 6 Human MDA-MB-231 breast cancer cells were orthotopically inoculated in the mammary pad of BALB / c nude mice until the tumor volume grew to about 100mm 3 . The thrombin-loaded DNA self-assembled nanoparticle solution prepared in Example 1 was injected into the tail vein of tumor-bearing mice at a dosage of about 1.5 U unit thrombin per mouse, and the tumor volume was measured once every 2 days. After 6 injections, the changes in tumor volume were statistically analyzed. Tumor volume was calculated according to the following formula: volume = (d 2 ×D) / 2, where d is the smallest diameter of the tumor and D is the largest diameter. Inject normal saline, free thrombin, and blank DNA nanoparticles as negative controls. Experimental results such as image 3 shown.

[0070] Depend on ...

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Abstract

The invention discloses a method for preparing a thrombin-loaded DNA (deoxyribonucleic acid) self-assembly nano-structure. The method comprises the following steps: self-assembling a single-stranded DNA molecule into a rectangular nano-structure by utilizing a DNA paper folding technology; immobilizing thrombin on the surface of the rectangular nano-structure, and immobilizing a 'lock' structure capable of answering vascular endothelial specific receptor of a tumor at two long edges of the rectangular nano-structure so as to close the thrombin inside the structure and generate response when thrombin is carried to a tumor blood vessel tissue. Therefore, target delivery and controllable release of the tumor blood vessel tissue by the thrombin can be realized; and the prepared thrombin-loaded DNA (deoxyribonucleic acid) self-assembly nano-structure can be used for intravenous injection, realizes targeted treatment of tumors, and has a wide clinical application prospect.

Description

technical field [0001] The invention relates to the technical field of nanocarrier medicines, in particular to a DNA self-assembled nanostructure loaded with thrombin, its preparation method and application. Background technique [0002] Tumor vasculature is closely related to tumor growth, invasion, and metastasis. It is not only the main place for tumor tissue to obtain nutrients and oxygen, but also the channel for tumors to discharge metabolites and tumor cells to escape and metastasize. good target. Tumor blood vessel-targeted therapy mainly includes two strategies: anti-angiogenesis therapy and tumor blood supply blocking therapy. Among them, blocking the tumor blood supply therapy is to embolize the existing blood vessels, so that the tumor cells will die due to lack of nutrients and oxygen starvation. The target of this treatment method is blood components, so it is not easy to produce drug resistance and has a wide range of applications. In addition, for most can...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61K9/14A61K38/48A61P35/00
Inventor 聂广军李素萍丁宝全宋晨田艳华蒋乔
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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