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Nano enzyme as well as preparation method and application thereof

A nano-enzyme and nano-particle technology, applied in the field of biomedical materials, can solve the problems of low utilization rate of small molecule drugs, large side effects, difficult to treat acute kidney injury, etc., and achieve excellent biocompatibility and biosafety, good effect of treatment

Active Publication Date: 2022-07-01
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a nanozyme and its preparation method and application, aiming at solving the problem that the existing small molecule drugs have low utilization rate, large side effects, and are difficult to be used in the treatment of acute kidney injury. question

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  • Nano enzyme as well as preparation method and application thereof
  • Nano enzyme as well as preparation method and application thereof
  • Nano enzyme as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Synthesis of Rh-Ser

[0044] Rh-Ser synthesis: 60 mg RhCl 3 ·3H 2O was dissolved in 50 mL of water, then 40 mg of methoxypolyethylene glycol mercapto was added to the solution with vigorous stirring. After 0.5 h, 5 mL of NaBH was quickly added dropwise to the mixture 4 Aqueous solution (2 mg / mL). After 10 minutes of reaction, the above solution was purified by ultrafiltration centrifugation. The sample was then freeze-dried to obtain a solid product. The dried solid product was then dispersed in water (2 mg / mL, 5 mL) and mixed with L-serine and stirred overnight to obtain Rh-Ser. Free L-serine was removed by ultrafiltration centrifugation and the purified solution was lyophilized for ready use. The threonine-modified rhodium nanozyme replaces serine with threonine, and the resulting sample is called Rh-Thr.

[0045] figure 1 is the route map for the synthesis of Rh-Ser, in which RhCl 3 ·3H 2 O stands for rhodium chloride trihydrate, and L-Ser stands...

Embodiment 2

[0047] Example 2: Rh-Ser has the ability to scavenge various reactive oxygen species / nitrogen.

[0048] The efficiency of scavenging hydroxyl radicals with different concentrations of Rh-Ser (0-100 μg / mL) was determined by hydroxyl radical antioxidant capacity (HORAC) kit (Cell Biolabs, USA). Testing was performed according to the protocol provided by the manufacturer.

[0049] like Figure 5 As shown, Rh-Ser can effectively scavenge hydroxyl radicals with concentration-dependent properties.

[0050] The superoxide anion scavenging efficiency of different concentrations of Rh-Ser (0-5 μg / mL) was determined by superoxide dismutase (SOD) detection kit (Sigma-Aldrich, USA). Testing was performed according to the protocol provided by the manufacturer.

[0051] like Image 6 As shown, Rh-Ser can effectively scavenge superoxide anion in a concentration-dependent manner.

[0052] Determination of Rh-Ser scavenging of 2,2'-azidobis(3-ethylbenzothiazoline-6-sulfonic acid)diammoniu...

Embodiment 3

[0058] Example 3: Rh-Ser cytotoxicity and protection of kidney cells by scavenging various reactive oxygen species / reactive nitrogen species The standard MTT method was used to evaluate the survival rate of Rh-Ser on 293T renal embryonic cells and human renal tubular epithelial cells (HK-2) Impact.

[0059] 293T cells or HK-2 cells at 1 × 10 per well 4 Density seeded into 96-well plates and placed at 37°C, 5% CO 2 Incubate for 12h under conditions. Next, the old medium in the 96-well plate was aspirated, and medium solutions containing different concentrations of Rh-Ser were added. After culturing for 20 or 44 hours, the old medium in the 96-well plate was aspirated, and 100 μL of MTT medium solution (0.8 mg / mL) was added to each well, and the culture was continued for 4 hours. Aspirate the residual medium in the 96-well plate, add 150 μL DMSO solution to each well, shake gently, detect the OD value of each well on a Synergy H1 microplate reader (detection wavelength is 570...

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Abstract

The invention discloses a nano-enzyme as well as a preparation method and application thereof, and the nano-enzyme comprises rhodium nano-particles and serine combined on the surfaces of the rhodium nano-particles. The nano-enzyme (Rh-Ser) provided by the invention has an ultra-small size which is lower than a renal tubule filtration threshold value; meanwhile, due to the fact that serine has the capacity of targeting renal injury molecularly 1 (KIM-1) expressed by damaged renal tubules, modification of serine can enable the rhodium nano-particles to have certain renal targeting capacity, and the rhodium nano-particles can be effectively enriched in the kidneys of damaged mice; a large amount of active oxygen or active nitrogen in the renal canaliculus is removed to relieve and treat glycerol-induced acute kidney injury, and the compound has excellent anti-inflammatory ability. In addition, the Rh-Ser has a good treatment effect, and meanwhile, the Rh-Ser has excellent biocompatibility and biological safety.

Description

technical field [0001] The invention relates to the technical field of biomedical materials, in particular to a nano-enzyme and a preparation method and application thereof. Background technique [0002] Acute kidney injury is an important human health problem with high morbidity and mortality. It is estimated that 1.7 million people worldwide die from acute kidney injury every year. Currently, adjuvant therapy and kidney transplantation are the most common treatments for acute kidney injury. Recent studies have shown that the pathogenesis of acute kidney injury is associated with excess intracellular reactive oxygen species and reactive nitrogen species. Previously, some small-molecule drugs, such as amifostine and acetylcysteine, have been shown to act as antioxidants to eliminate reactive oxygen species, thereby alleviating acute kidney injury. However, the existing small-molecule drugs are not targeted, so the utilization rate is low, and they are prone to cause large ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K33/24A61K47/54A61P13/12A61P39/06B82Y5/00B82Y40/00
CPCA61K47/542A61K33/24B82Y5/00B82Y40/00A61P13/12A61P39/06
Inventor 黄鹏涂天慧张东阳林静
Owner SHENZHEN UNIV
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