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Construction method and application of a dual-target DNA nanotherapy system

A construction method and dual-target technology, applied in the field of biomedicine, to achieve the effect of stable structure and simple synthesis method

Active Publication Date: 2021-06-04
ZHENGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The invention proposes a construction method and application of a dual-target DNA nanotherapy system, which solves the technical problems existing in traditional drug delivery systems

Method used

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Examples

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Embodiment 1

[0027] A method for constructing a dual-target DNA nanotherapy system in this embodiment:

[0028] (1) Synthesis of DNA tetrahedron:

[0029] Mix the six single strands S1, S2, S3, S4, S5, and S6 required for the synthesis of DNA tetrahedron in equimolar proportions, and add Tris∙HCl (pH 8.0) and MgCl 2 Buffer to make single chain final concentration 0.5 μM, Tris∙HCl (pH 8.0) final concentration 10 mM, MgCl 2 The final concentration is 5mM, vortex at low speed to make it evenly mixed. The sample was placed in a PCR instrument, denatured at 95°C for 5 minutes, then rapidly cooled to 4°C and kept for 30s. The blank DNA tetrahedron was synthesized and stored at 4°C.

[0030] (2) Synthesis of ssDNA-peptide

[0031] Add cross-linking agent Sulfo-SMCC powder to the ssDNA solution modified with C6 amino group at the 5' end, so that the molar ratio of the two substances is 1:110, pH 7~9, under magnetic stirring, react at room temperature for 2 hours, and dialyze for 24 hours to re...

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Abstract

The invention belongs to the technical field of biomedicine, in particular to a construction method and application of a dual-target DNA nanotherapy system. Dissolve six single-stranded linear DNAs in in vitro buffered saline solution, anneal and self-assemble to form a DNA tetrahedron solution; add cross-linking agent Sulfo-SMCC powder to the ssDNA solution modified with C6 amino group at the 5' end, react at room temperature for 2 hours, remove untreated The reacted cross-linking agent is then added to the peptide powder, reacted at room temperature for 12 hours, and the excess peptide is removed to obtain ssDNA-peptide; ssDNA-peptide is added to the DNA tetrahedron solution to complete the construction of the dual-target DNA nanotherapy system. The structure of the DNA tetrahedron carrier constructed by the present invention is stable, and the stability of the polypeptide drug can be improved after the carrier is loaded with drugs, and the dual-target DNA nanotherapy system has strong targeting, and mainly plays a biological therapeutic role by regulating cell signaling pathways.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a construction method and application of a dual-target DNA nanotherapy system. Background technique [0002] DNA nanostructures have the advantages of passive targeting into the lesion area, overcoming multidrug resistance, and increasing the residence time of drugs in target cells. Among many DNA nanostructures, the synthesis method of DNA tetrahedron is simple, and the DNA materials used are less , strong structural rigidity, and easy to modify. [0003] Fas (CD95) is a type I transmembrane (TM) protein on the cell surface, and it is a prototype member of the death receptor (DR) family in the tumor necrosis factor receptor (TNFR) superfamily. FasL binds to induce tumor cell apoptosis by regulating related signaling pathways. [0004] Studies have found that due to the increased exposure of negatively charged phosphatidylserine on the surface of cancer cells, antimicrobial...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/54A61K47/62A61K47/66A61K47/64A61K38/10A61K38/16A61P31/04A61P35/00
CPCA61K38/10A61K38/16A61K47/549A61K47/62A61K47/64A61K47/66A61P31/04A61P35/00
Inventor 赵永星张楠杨亚楠杨静
Owner ZHENGZHOU UNIV
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