Preparation method of NF-kB decoy ODNs carried by targeting DNA nano-self-assembly body and application thereof

A nanometer self-assembly and targeting technology, applied in the field of medicine, can solve the problems of poor anti-inflammatory effect, poor stability, and large side effects, and achieve the effect of enhancing anti-inflammatory effect, improving stability and small side effects.

Active Publication Date: 2017-11-10
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In order to solve the technical problems of poor stability, poor anti-inflammatory effect and large side effects of NF-κB decoy ODNs in the treatment of

Method used

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  • Preparation method of NF-kB decoy ODNs carried by targeting DNA nano-self-assembly body and application thereof
  • Preparation method of NF-kB decoy ODNs carried by targeting DNA nano-self-assembly body and application thereof
  • Preparation method of NF-kB decoy ODNs carried by targeting DNA nano-self-assembly body and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Synthesis of NF-κB decoy ODNs:

[0033] The two single-chain NF-κB D and NF-κB D' of the synthesized NF-κB decoy ODNs were mixed in an equimolar ratio, diluted with TM (Tris∙HCl, pH 8.0 and MgCl2) buffer to a final concentration of 0.5 μM, Vortex at low speed until well mixed. In a PCR instrument, the temperature was lowered from 80 °C to 25 °C for 2 h. NF-κB decoy ODNs were synthesized and stored at 4 ℃.

[0034] (2) Synthesis of DNA nano-self-assembly:

[0035] Mix the four single-strands S1, S2, S3, and S4 of the synthetic DNA nano-self-assembly in equimolar ratios, dilute with 5mM TM (Tris∙HCl, pH 8.0 and MgCl2) buffer to a final concentration of 0.5 μM, and vortex at low speed until well mixed. In a PCR instrument, after denaturation at 95°C for 5 min, the temperature was rapidly lowered to 4°C and maintained for 30 s. DNA nano-self-assembly was synthesized and stored at 4°C.

[0036] (3) Synthesis of NF-κB decoy ODNs loaded with DNA nano-self-assembly: ...

Embodiment 2

[0045] (1) Synthesis of NF-κB decoy ODNs:

[0046] The two single-chain NF-κB D and NF-κB D' of the synthesized NF-κB decoy ODNs were mixed in an equimolar ratio, diluted with TM (Tris∙HCl, pH 8.0 and MgCl2) buffer to a final concentration of 0.5 μM, Vortex at low speed until well mixed. In a PCR instrument, the temperature was lowered from 80 °C to 25 °C for 2 h. NF-κB decoy ODNs were synthesized and stored at 4 ℃.

[0047] (2) Synthesis of DNA nano-self-assembly:

[0048] Mix the four single-strands S1, S2, S3, and S4 of the synthetic DNA nano-self-assembly in equimolar ratios, dilute with 5mM TM (Tris∙HCl, pH 8.0 and MgCl2) buffer to a final concentration of 0.5 μM, and vortex at low speed until well mixed. In a PCR instrument, after denaturation at 95°C for 5 min, the temperature was rapidly lowered to 4°C and maintained for 30 s. DNA nano-self-assembly was synthesized and stored at 4°C.

[0049] (3) Synthesis of NF-κB decoy ODNs loaded with DNA nano-self-assembly: ...

Embodiment 3

[0058] (1) Synthesis of NF-κB decoy ODNs:

[0059] The two single-chain NF-κB D and NF-κB D' of the synthesized NF-κB decoy ODNs were mixed in an equimolar ratio, diluted with TM (Tris∙HCl, pH 8.0 and MgCl2) buffer to a final concentration of 0.5 μM, Vortex at low speed until well mixed. In a PCR instrument, the temperature was lowered from 80 °C to 25 °C for 2 h. NF-κB decoy ODNs were synthesized and stored at 4 ℃.

[0060] (2) Synthesis of DNA nano-self-assembly:

[0061] Mix the four single-strands S1, S2, S3, and S4 of the synthetic DNA nano-self-assembly in equimolar ratios, dilute with 5mM TM (Tris∙HCl, pH 8.0 and MgCl2) buffer to a final concentration of 0.5 μM, and vortex at low speed until well mixed. In a PCR instrument, after denaturation at 95°C for 5 min, the temperature was rapidly lowered to 4°C and maintained for 30 s. DNA nano-self-assembly was synthesized and stored at 4°C.

[0062] (3) Synthesis of NF-κB decoy ODNs loaded with DNA nano-self-assembly: ...

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Abstract

The invention belongs to the technical field of medicine, and relates to a preparation method of NF-kB decoy ODNs carried by a targeting DNA nano-self-assembly body and application thereof. NF-kB decoy ODNs carried by the targeting DNA nano-self-assembly body is composed of a DNA tetrahedron, NF-kB decoy ODNs and VCAM-1 targeting polypeptide, wherein NF-kB decoy ODNs and VCAM-1 targeting polypeptide are connected to the DNA nano-self-assembly body according to the rule of complementation base pairing. The average particle size of the DNA nano-self-assembly body is less than 10 nm. NF-KB decoy ODNs carried by the targeting polypeptide-modified DNA nano-self-assembly body improves the targeting capability of genetic drugs and the in-vitro anti-inflammatory effect of the drugs, and has no toxicity to cells and tissue.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a preparation method and application of targeted DNA nanometer self-assembled NF-κB decoy ODNs. Background technique [0002] The nuclear factor-κB / Rel transcription factor family plays a central role in coordinating the expression of several genes involved in immune and inflammatory processes. NF-κB is expressed in the cytoplasm of almost all cells. Normally, NF-κB is usually It is a heterodimer of inhibitory protein (IκB) combined with p50 and p65 subunits, but in the relevant cells of diseases involved in inflammation, the cells are activated by various factors, such as pro-inflammatory cytokines, bacterial lipopolysaccharide (LPS ) and phorbol ester, etc., when NF-κB is activated, the inhibitor of NF-κB, IκB, will be phosphorylated and degraded, and release the P50 / P65 complex that is separated from IκB. Transcription and expression of inflammatory genes. Gene drug NF-κB dec...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/54A61K47/64A61P29/00A61P19/02
CPCA61K48/0025A61K48/005
Inventor 赵永星张楠华海婴付玲玲
Owner ZHENGZHOU UNIV
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