Construction method and application of double-target-spot DNA (Deoxyribonucleic Acid) nano therapy 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: 2018-11-02
ZHENGZHOU UNIV
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Problems solved by technology

[0006] The invention proposes a construction method and application of a dual-target DNA nan...

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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 biomedicines and in particular relates to a construction method and application of a double-target-spot DNA (Deoxyribonucleic Acid) nano therapy system.The construction method comprises the following steps: dissolving six single-stranded linear-chain DNA into an in-vitro buffer salt solution; carrying out annealing and self-assembling to form a DNAtetrahedron solution; adding crosslinking agent Sulfo-SMCC powder into a 5'-end C6 amino modified ssDNA solution; reacting at room temperature for 2h; removing an unreacted crosslinking agent and thenadding polypeptide powder; reacting at the room temperature for 12h and removing excessive polypeptide to obtain ssDNA-peptide; adding the ssDNA-peptide into the DNA tetrahedron solution, so as to finish construction of the double-target-spot DNA nano therapy system. According to the construction method provided by the invention, a constructed DNA tetrahedron carrier has a stable structure; aftera medicine is loaded on the carrier, the stability of a polypeptide medicine can be improved; the double-target-spot DNA nano therapy system has relatively strong targeting performance and mainly express a biological therapy effect through regulating and controlling a cell signal path.

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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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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