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Method for promoting arginine to quickly release NO by black phosphorus

A technology of arginine and black phosphorus, which is applied in the directions of biochemical equipment and methods, pharmaceutical formulations, medical preparations with inactive ingredients, etc., can solve the problems of slow release and low release amount, etc. Good stability and good photothermal effect

Active Publication Date: 2020-11-03
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] As a NO donor, arginine has been used in various antibacterial and antitumor treatments, but there are defects such as slow release and low release in this process, so an effective strategy is urgently needed to make up for the existing deficiencies

Method used

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  • Method for promoting arginine to quickly release NO by black phosphorus
  • Method for promoting arginine to quickly release NO by black phosphorus
  • Method for promoting arginine to quickly release NO by black phosphorus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Synthesis and Characterization of BP-Arg-GOx

[0032] (1) Synthesis of BP-Arg: Measure 2 mL of 1 mg / mL black phosphorus in DMF (or dimethyl sulfoxide) solution, add 6 mg of arginine (you can also use canavanic acid and other guanidine groups that can be oxidized to produce NO) compound) and 3 mg DMAP, and then 200 μL of deionized water was added, and the mixture was reacted overnight for 8 h. After the reaction was completed, the mixed solution was dialyzed for 6 hours to remove unreacted arginine. After the dialysis, the obtained product was centrifuged at 17,000 g for 4 minutes, washed three times with deionized water, and finally the obtained product was resuspended in deionized water for storage.

[0033] (2) Synthesis of BP-Arg-GOx: Weigh 2mg GOx, add 5mg EDC and 10mg NHS, activate the carboxyl group on GOx in PB, add 5mg BP-Arg to the activated GOx after 30min, react overnight, centrifuge Take the pellet and resuspend it in PB for subsequent use.

[0034] BP-Arg...

Embodiment 2

[0036] Determination of particle size spectrum: Take appropriate amount of black phosphorus and BP-Arg-GOx respectively, and measure their hydrodynamic diameters in PB 7.4 respectively. In addition, the hydrodynamic diameter of BP-Arg was measured on the 1st, 5th, and 10th day to monitor its stability.

Embodiment 3

[0038] Determination of NO release profile: take a certain amount of BP-Arg-GOx, and measure its NO release.

[0039] as attached Figure 1-4 As shown, the modified product BP-Arg-GOx and the raw material BP were analyzed by TEM and DLS. The surface of the modified BP showed a significant difference from the raw material BP. The DLS results also proved that BP-Arg-GOx compared with BP , the particle size increases to a certain extent.

[0040] as attached Figure 5 As shown, the DLS analysis of the modified product BP-Arg-GOx shows that BP-Arg-GOx still maintains good stability after 5 days in PBS solution.

[0041] as attached Figure 6 As shown, the modified product BP-Arg-GOx showed similar enzyme activity to free GOx after NIR irradiation, indicating the success of BP-Arg-GOx modification and the maximum retention of enzyme activity.

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Abstract

The invention discloses a method for promoting arginine to quickly release NO by black phosphorus. The method comprises the following steps: taking 4-dimethylaminopyridine (DMAP), a guanidyl-containing compound and the black phosphorus, and performing reacting in a mixed solution of an organic solvent and water; after the reaction is finished, removing the unreacted guanidyl-containing compound; centrifugally separating the reacted dispersion liquid to obtain protected black phosphorus (BP- guanidyl-containing compounds); taking 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), N-hydroxysuccinimide (NHS) and glucose oxidase (GOx), and performing reacting in PB at normal temperature; adding the BP-guanidyl-containing compounds into the reaction in the previous step, and performing centrifuging after the reaction to obtain GOx and a BP-guanidyl-containing compound conjugate (BP-guanidyl-containing compound-GO<x>); and dissolving a BP-guanidyl-containing compound- GOx in a glucose solution, irradiating the black phosphorus with near-infrared light to promote the GOx to generate hydrogen peroxide (H2O2), and oxidizing the guanidyl-containing compound with H2O2 to quickly generate NO. According to the method, the black phosphorus photothermal effect of the black phosphorus is utilized to promote glucose oxidase to generate hydrogen peroxide and promote arginine oxidation to quickly generate the NO. The obtained covalently protected black phosphorus is good in stability, high in enzyme activity, rapid in NO release and high in yield.

Description

technical field [0001] The invention relates to a preparation method of a two-dimensional material, in particular to a method for promoting fast release of NO from arginine by black phosphorus. Background technique [0002] Nitric oxide (NO) is an important biological signal molecule and effector molecule in organisms, which is produced by NO synthase (NOS) catalyzed by arginine, and plays various physiological roles in various tissues in organisms such as expansion Blood vessels, anti-tumor, etc., but limited by the amount of endogenous NO production, it is often difficult to meet the needs of the body to cope with lesions, so exogenous NO donors have been developed as their main source. The types of NO donors are mainly divided into organic nitrates, S-nitrosothiols, nitroprussides, furazan N-oxides, azonium dialkoxides, etc. These small molecule NO donors have been widely used in the treatment of diseases, but they also have the disadvantages of short half-life, poor spe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61K38/44A61K47/54A61K47/55A61P35/00A61P9/08B82Y5/00
CPCA61K41/0052A61K38/443A61K47/542A61K47/55A61P35/00A61P9/08C12Y101/03004B82Y5/00A61K2300/00
Inventor 乔海石黄鑫陈维
Owner CHINA PHARM UNIV
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