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Veterinary preparation namely lysozyme-loaded chitosan microspheres and preparation method thereof

A technology of chitosan microspheres and lysozyme, which is used in antibacterial drugs, pharmaceutical formulations, medical preparations containing active ingredients, etc., to achieve the effects of enhancing inhibitory ability, reducing difficulty and mild reaction conditions

Inactive Publication Date: 2015-03-11
NANJING HAOYU AGRI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the veterinary preparations that enhance the inhibitory effect of lysozyme on Gram-negative bacteria (G-) through the auxiliary effect of chitosan, and the related preparation methods have not been reported.

Method used

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  • Veterinary preparation namely lysozyme-loaded chitosan microspheres and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Add chitosan hydrochloride with a molecular weight of 5kDa and a degree of deacetylation>85% into water, stir and dissolve at room temperature, and filter through micropores to obtain a chitosan aqueous solution with a concentration of 5.0mg / ml. Then add lysozyme with an activity of 20000U / mg, so that its concentration in the chitosan aqueous solution is 0.1mg / ml. With dextran sulfate as the negatively charged cross-linking agent, the aqueous solution of dextran sulfate is added dropwise to the aqueous chitosan solution containing lysozyme at a rate of 7.0ml / h using a medical syringe pump, and the stirring of the aqueous chitosan solution at this time The speed was 700 rpm, and the stirring was continued for 30 min after the dropwise addition was completed, so as to obtain a suspension of chitosan microspheres loaded with lysozyme. The suspension was allowed to settle naturally for 6 hours, and the upper milky white suspension was separated. Centrifuge at 4°C (at a rat...

Embodiment 2

[0028] Add chitosan hydrochloride with a molecular weight of 20kDa and a degree of deacetylation>85% into water, stir and dissolve at room temperature, and filter through micropores to obtain a chitosan aqueous solution with a concentration of 1.0mg / ml. Then add lysozyme with an activity of 20000U / mg, so that its concentration in the chitosan aqueous solution is 0.8mg / ml. With dextran sulfate as the negatively charged cross-linking agent, the aqueous solution of dextran sulfate is added dropwise to the aqueous solution of chitosan containing lysozyme at a rate of 2.5ml / h using a medical syringe pump. The speed was 1000 rpm, and the stirring was continued for 30 min after the dropwise addition was completed, so as to obtain a suspension of chitosan microspheres loaded with lysozyme. The suspension was allowed to settle naturally for 6 hours, and the upper milky white suspension was separated. Centrifuge at 4°C (at a rate of 8500xg) for 30min, remove the sediment from the lower...

Embodiment 3

[0031]Add chitosan hydrochloride with a molecular weight of 20kDa and a degree of deacetylation>85% into water, stir and dissolve at room temperature, and filter through micropores to obtain a chitosan aqueous solution with a concentration of 1.5mg / ml. Then add lysozyme with an activity of 20000U / mg, so that its concentration in the chitosan aqueous solution is 0.8mg / ml. With sodium alginate as the negatively charged cross-linking agent, the aqueous solution of dextran sulfate is added dropwise to the aqueous chitosan solution containing lysozyme at a rate of 4.5ml / h using a medical syringe pump. At this time, the stirring rate of the aqueous chitosan solution is 300rpm, continue to stir for 10min after the dropwise addition, to obtain the chitosan microsphere suspension loaded with lysozyme. The suspension was allowed to settle naturally for 6 hours, and the upper milky white suspension was separated. Centrifuge at 4°C (at a rate of 8500xg) for 30min, remove the sediment fro...

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Abstract

The invention belongs to the field of veterinary preparation, and relates to a veterinary preparation namely lysozyme-loaded chitosan microspheres and a preparation method thereof. The chitosan with a molecular weight of 5 to 50 kDa is crosslinked with an electronegative crosslinking agent through the electrostatic action so as to form microspheres, and the lysozyme is encapsulated in the microspheres. The ion-crosslinking method and the freeze-drying method are co-used to prepare the veterinary preparation, the reaction conditions are mild, the technology process is controllable, the industrialization development becomes easier, and thus the preparation method is practical. The lysozyme and chitosan can generate a synergetic effect, the chitosan with a molecular weight of 5-50 kDa can penetrate the outer membrane of G- cells, and thus the lysozyme can be transferred to the inner wall layer of G- cells to hydrolyze the peptidoglycan structure, so that the inhibiting performance of lysozyme on Gram-negative bacteria (G-) is effectively strengthened, and the preparation can partially replace the broad-spectrum antibacterial effect of antibiotics.

Description

technical field [0001] The invention belongs to the field of veterinary drug preparations, and relates to a veterinary drug preparation loaded with chitosan microspheres and a preparation method thereof. Background technique [0002] Under the current breeding environment, diseases caused by bacterial infections in livestock and poultry are on the rise, and the prevention and control methods mainly rely on antibiotics. Due to long-term irrational use, animal-derived bacteria have developed serious resistance to antibiotics, which has also aroused people's concerns about food safety. It is imminent to find alternatives to antibiotics with high efficiency and broad spectrum. At present, the innovative research on veterinary antibacterial drugs is mainly based on the following aspects: (1) Chemically improve the parent structure or pharmacophore of antibiotics to develop a new generation of antibiotics for the drug resistance mechanism of bacteria. However, from the perspectiv...

Claims

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Application Information

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IPC IPC(8): A61K38/47A61K9/16A61P31/04A61K31/722
Inventor 余刚秦竹林家彬周忠凯李辉唐玉新汤赤陈俊
Owner NANJING HAOYU AGRI TECH
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