Kidney-targeting DNA nanometer raft-IL-33 as well as preparation method and application thereof

A kidney-targeted, SH-DNA technology, applied in the field of kidney-targeted DNA nano-raft-IL-33 and its preparation, can solve problems such as complexity, poor accumulation, poor cytokine targeting performance, etc. Achieve the effects of relieving pain, reducing drug costs, and improving drug targeting efficiency

Pending Publication Date: 2021-12-24
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, exosomes are primarily targeted to and enriched in the liver, leading to suboptimal accumulation of cytokine-encapsulated exosomes in the kidney
The technology of us

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  • Kidney-targeting DNA nanometer raft-IL-33 as well as preparation method and application thereof
  • Kidney-targeting DNA nanometer raft-IL-33 as well as preparation method and application thereof
  • Kidney-targeting DNA nanometer raft-IL-33 as well as preparation method and application thereof

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Example Embodiment

[0037] The present invention discloses a cytokine-based chemical synthesis method based on DNA nanobire, including the following steps:

[0038] (1) Preparation: Prepare DNA Single Chain (M 13MP 18DNA single strand, staple chain, capture chain), mercapto-modified DNA (SH-DNA), recombinant interleukin 33, 3- (2-pyridyl dithiosis) N-hydroxy succinimide (SPDP), 1XTAE-MG 2+ Buffer.

[0039] (2) Preparation of DNA-IL-33, separated by ultrafiltration tubes, and produces preparation efficiency by ultraviolet-visible spectroscopy;

[0040] (3) Synthesis of DNA origami rafts, with a capture chain complementary to DNA in step (1) by PCR synthesis. The morphology is characterized by the ultrafiltration tube separation, and its morphology is characterized by the atomic force microscope (AFM).

[0041] (4) Assembly synthesis of DNA nanoptrin, mixing the DNA-IL-33 and step (2) of step (1), and assembled DNA nanofibran, assembled, separated by ultrafiltration tube, characterized by AFM Morality,...

Example Embodiment

[0062] Example 1 Preparation of DNA-IL-33

[0063] 100 μg of IL-33 powder was dissolved in 50 μl of NaCl solution. 50 μl of 6.6 μm IL-33 solution was reacted for 2 hours in 1X PBS buffer (pH 8.5) in 1X PBS buffer (pH 8.5). Excess SPDP was removed by Zeba rotary dehydrate column (7 kmWco, 0.5 mL). Then, IL-33 and mercapto-modified DNA chains (excess 3 times) were counted at room temperature at room temperature in 1 xpbs buffer (pH 7.4). The efficiency of HS-DNA modified IL-33 was evaluated by monitoring the net increase of the UV-VIS spectrum due to the formation of pyridin-2-thionone (pad coefficient: 8080 m-1 cm-1) at 343 nm. In 1X PBS (pH = 7.4), the excess SH-DNA chain was removed with a 30 kD cut filter (Amicon) (3000 g, 10 minutes).

Example Embodiment

[0064] Example 2 Synthesis of DNA origami

[0065] In 1x Tae-Mg 2+ In buffer, 10 nm M 13 MP 18 DNA single strands were mixed with 50 times excess staple nail chain and 100 times excess capture. The mixture was annealed from 95 ° C to 4 ° C.

[0066] Table 3 Annealed procedure from 95 ° C to 4 ° C

[0067] temperature gradient 95℃ 2min 95-60℃ -0.1℃ / 12s 60℃ 12min 60-25℃ -0.1℃ / 12s 4℃ Keep

[0068] At 1XTAE-MG 2+ Ultrafiltration (3000 g, 10 minutes) was permeated (3000 g, 10 minutes) twice in buffer, and then ultrafiltrated once (3000 g, 10 minutes) in 1X PBS (pH = 7.4), and the excess staple chain was removed. And capture the chain and characterize the atomic force microscope (AFM).

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Abstract

The invention relates to the field of biological medicines, in particular to a kidney-targeting DNA nanometer raft-IL-33 as well as a preparation method and application thereof. According to the kidney-targeting DNA nanometer raft-IL-33 as well as the preparation method and application thereof, a DNA nanometer technology is adopted to accurately and quantitatively load the IL-33 on a DNA nanometer raft, such that the drug use amount and the drug administration frequency of the IL-33 are reduced, the drug cost is reduced, the pain caused by multiple drug administration is reduced, the drug targeting efficiency is improved, and the side effect is reduced.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the kidney-targeted DNA nano-raft-IL-33 and its preparation method and application. Background technique [0002] Numerous studies have shown that the cytokine interleukin 33 (IL-33) has a positive role in the prevention of ischemic acute kidney injury (AKI) and renal protection. The IL-33-induced increase in ILC2 prompts ILC2 to produce large amounts of Th2 cytokines (IL-4 and IL-13), thereby suppressing the pro-inflammatory activity of the type 1 immune response (TNF-α, IL-1b, IL-6, CXCL1 and CXCL2 significantly decreased). In addition, IL-33-induced ILC2 directly promotes tubule repair by producing Areg to attenuate renal injury. IL-33 can also promote the increase of anti-inflammatory macrophages (M2). M2 produces anti-inflammatory cytokines and secretes HO-1 anti-inflammatory enzyme, thereby inhibiting inflammation and tissue damage. IL-33 has a positive preventive and protect...

Claims

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Application Information

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IPC IPC(8): A61K47/69A61K47/54A61K38/20A61P13/12B82Y5/00B82Y30/00B82Y40/00
CPCA61K38/20A61K47/549A61K47/6949A61P13/12B82Y5/00B82Y30/00B82Y40/00
Inventor 李威
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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