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Preparation method and application of dual-targeting DNA nano drug-loaded complex

A nano-drug-loading and complex technology is applied in the field of biomedicine to achieve the effect of strong resistance to nuclease degradation and important application prospects

Pending Publication Date: 2021-12-10
QUANZHOU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, the above-mentioned targeted therapy strategies all only aim at a single target on the cell surface to realize the recognition of tumor cells.

Method used

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  • Preparation method and application of dual-targeting DNA nano drug-loaded complex
  • Preparation method and application of dual-targeting DNA nano drug-loaded complex
  • Preparation method and application of dual-targeting DNA nano drug-loaded complex

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Experimental program
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Effect test

Embodiment 1

[0054] Embodiment 1 Materials and methods

[0055] The cell line used in the present invention is human malignant melanoma cell A375, purchased from China Center for Type Culture Collection. The single-stranded DNA used to construct the DNA tetrahedron in the present invention was purchased from the Shanghai Synthesis Department of Shanghai Sangong. The specific sequence, modification method and modification site are shown in Table 1 below.

[0056] Table 1 DNA single-strand sequence and modification method

[0057]

[0058] The nucleic acid aptamers involved in the present invention were all purchased from the Shanghai Synthesis Department of Shanghai Sangon. The specific sequences, modification methods and modification sites are shown in Table 2 below.

[0059] Table 2 Nucleic acid aptamer sequence and modification method

[0060]

[0061] 1.1 The preparation of the solution involved in the present invention

[0062] 1) 10% complete cell culture medium: Take 180mL of ...

Embodiment 2

[0096] Embodiment 2 experimental result

[0097] Construction of the first part DNA nano drug-loaded complex

[0098] 1. Construction of single-targeting and dual-targeting DNA nano drug-loaded complexes

[0099] 1.1 Construction of DNA tetrahedron

[0100] In the present invention, similar to the DNA tetrahedron structure reported in the literature, although the present invention adds an arm chain with a sticky end to the DNA tetrahedron, the design of the sticky end is based on the premise that the spatial structure of the DNA tetrahedron is not affected. The constructed "DNA tetrahedron-Aptamer-AS1411" has a special spatial structure, and the commonly used markers are not accurate. We increase the DNA single strands one by one by PCR, mutually verify the increase in the molecular weight of the nucleic acid chain during the construction process, and verify the construction by distinguishing the molecular weight change with the accuracy of PAGE gel. It can be seen from the...

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Abstract

The invention discloses a preparation method and application of a dual-targeting DNA nano drug-loaded complex. DNA single chains S1, S2, S4, S5, S7 and S8, a nucleic acid aptamer 1, a natural nucleic acid aptamer AS1411 and chemically modified nucleic acid four ligands AS1411-1, AS1411-2 and AS1411-3 are mixed in a TM buffer solution, the mixture is placed in a PCR instrument to be constructed, and a DNA tetrahedral Aptamer 1-AS1411, a DNA tetrahedral Aptamer 1-AS1411-1, a DNA tetrahedral Aptamer 1-AS1411-2 and a DNA tetrahedral Aptamer 1-AS1411-3 are obtained through construction respectively. According to the preparation method and application of the dual-targeting DNA nano drug-loaded complex, four dual-targeting DNA nano drug-loading systems based on CD44 and nucleolin nucleic acid aptamers are constructed, and compared with a single-targeting drug-loading complex and a negative control group, four dual-targeting drug-loaded complexes have stronger nuclease degradation resistance, capability of targeting melanoma A375 cells, capability of inhibiting proliferation of the A375 cells and capability of inducing the A375 cells to generate G1 phase retardation.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a preparation method and application of a dual-target DNA nano drug-carrying complex. Background technique [0002] Melanoma (Malignant melanoma) is one of the most malignant human skin tumors. In recent years, the incidence of melanoma has continued to increase and is showing a younger trend. Although recent advances in immunotherapy and molecularly targeted therapy have provided some more effective treatment modalities for melanoma, these treatments still cannot completely solve the problem of drug toxicity. Therefore, developing new targeted therapies for melanoma and reducing the side effects of chemotherapy drugs are crucial to improving the overall efficacy of melanoma treatment. [0003] CD44 molecule is a kind of transmembrane single-chain glycoprotein, which regulates various biological functions of cells by mediating signal transduction between cells and between cells and ex...

Claims

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Application Information

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IPC IPC(8): A61K47/54A61K47/64A61K31/704A61P35/00B82Y5/00B82Y40/00
CPCA61K47/549A61K47/64A61K31/704A61P35/00B82Y5/00B82Y40/00
Inventor 孙丽丹谢晓兰吕凤娇
Owner QUANZHOU NORMAL UNIV
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