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Application of ruthenium complex serving as nucleic acid vector of target cell nucleus

A technology of ruthenium complex and nucleic acid carrier is applied in the new application field of ruthenium complex as nucleic acid carrier, and can solve the problems of gene carrier application of ruthenium complex not being reported to target cell nucleus, large molecular weight of ruthenium complex and limited effect, etc.

Active Publication Date: 2016-01-13
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Kumbhar et al. reported an application of ruthenium polypyridyl complexes as gene carriers (S.S.Bhat, A.S.Kumbhar, A.K.Kumbhar, A.Khan, P.Lonnecke, Rutheium(II) polypyridylcomplexes as carriers for DNAdelivery, Chem.Comm., 2011,47, 11068-1070); Chao Hui et al reported the method of polypyridine ruthenium (II) complex as gene carrier (B.Yu, Y.Chen, C.Ouyang, H.Huang, L.JiandH.Chao; Aluminescenttetranuclearruthenium (II ) complex as a tracking non-viral gene vector, Chem.Commun., 2013,49,810-812). However, the molecular weight of the ruthenium complex used in these reports is relatively large, the synthesis process is relatively complicated, and its effect as a gene carrier is very limited, and the above reports Ruthenium complexes and their optical isomers have not been reported as gene carriers targeting the nucleus

Method used

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  • Application of ruthenium complex serving as nucleic acid vector of target cell nucleus
  • Application of ruthenium complex serving as nucleic acid vector of target cell nucleus
  • Application of ruthenium complex serving as nucleic acid vector of target cell nucleus

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1:[Ru(bpy)pBEPIP](ClO 4 ) 2 － c-myc booting section dna Preparation of complex

[0042] Experimental method: 1mM [Ru(bpy) 2 pBEPIP] (ClO 4 ) 2 ( figure 1 A) Mix evenly with 1mMc-mycPu22 solution at a ratio of 1:1, heat to 90°C for 0.5-10min, then naturally cool to room temperature, and stand at 4°C for 24 hours. The reaction mixture was dialyzed with distilled water to remove the polypyridine ruthenium (II) complex that was not loaded into the nucleic acid, the polypyridine ruthenium (II) complex-nucleic acid complex, and TEM detection found that the ruthenium complex promoted the c-mycPu22 sequence self-assembled into nanotubes ( figure 1 B).

Embodiment 2

[0043] Example 2:[Ru(bpy) 2 pBEPIP] (ClO 4 ) 2 - telomere dna Preparation of complex

[0044] Experimental method: 1mM [Ru(bpy) 2 pBEPIP] (ClO 4 ) 2 Mix evenly with 1mM telomere DNA (5'-TTAGGG-3') solution at a ratio of 1:1, heat to 90°C for 0.5-10min, then naturally cool to room temperature and let stand at 4°C 24 hours. The reaction mixture was dialyzed with distilled water to remove the polypyridine ruthenium (II) complex that was not loaded into the nucleic acid, the polypyridine ruthenium (II) complex-nucleic acid complex, and TEM detection found that the ruthenium complex promoted the telomeric DNA sequence self-assembled into a nanotube-like structure ( figure 2 ).

Embodiment 3

[0045] Example 3:[Ru(bpy) 2 pBEPIP] (ClO 4 ) 2 and bcl-2 booting section dna Preparation of complex

[0046] Experimental method: 1mM [Ru(bpy) 2 pBEPIP] (ClO 4 ) 2 Mix evenly with 1mMBcl-2pu27 solution at a ratio of 1:1, heat to 90°C for 0.5-10min, then naturally cool to room temperature, and stand at 4°C for 24 hours. The reaction mixture was dialyzed with distilled water to remove the polypyridine ruthenium (II) complex that was not loaded into the nucleic acid, the polypyridine ruthenium (II) complex-nucleic acid complex, and TEM detection found that the ruthenium complex promoted the telomeric DNA sequence self-assembled into a nanotube-like structure ( image 3 ).

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Abstract

The invention discloses application of a ruthenium complex serving as a nucleic acid vector of a target cell nucleus. According to experimental data, the ruthenium (II) complex can be effectively combined with nucleic acid sequences and effectively change forms of long-sequence nucleic acids to enable the nucleic acids to be effectively transported into living cells in a transmembrane manner and well positioned in cell nucleuses, so that nucleic acid transporting efficiency is greatly improved. Therefore, the nucleic acid sequences can be conveniently transferred into the cells to realize gene therapy, fluorescent staining tracing or the like. By a ruthenium complex-nucleic acid compound preparation method, stable ruthenium complex-nucleic acid compounds can be prepared efficiently, and better effects are achieved.

Description

technical field [0001] The invention relates to a new application of the ruthenium complex, in particular to a new application of the ruthenium complex as a nucleic acid carrier. Background technique [0002] The treatment of tumor and other diseases by gene therapy technology has been paid more and more attention by researchers. The so-called gene therapy refers to the transfection (transfection) of certain functional genetic materials, including siRNA, mRNA, miRNA, DNA, nucleic acid aptamers, and promoter regions of cancer genes, into cells so that they can be expressed in cells, and eventually To achieve the purpose of treating diseases. The key to gene therapy is to transfect therapeutic genes into cells with safe and efficient vectors. Viral (viral) vectors such as RNA virus or DNA virus, or non-viral (non-viral) vectors, including calcium phosphate precipitation, lipofection, microinjection, etc., can be used to transfect therapeutic genes (H.Yin ,R.L.Kanasty,A.A.El...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C09K11/06C12Q1/68C12N15/11
CPCC09B57/10C09B69/008C12N15/87C12Q1/6806C07F15/0053C12Q2563/137C09B57/00C12Q1/68
Inventor 梅文杰
Owner GUANGDONG PHARMA UNIV
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