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Hybrid protein nano oxygen carrier and its preparation method and application

A technology of hybrid protein and oxygen carrier, which is applied in the field of oxygen carrier, can solve problems such as difficulty in maintaining the functional stability of hemoglobin, mismatch of blood type, and restrictions on the application of emergency and severe patients, and achieve good biocompatibility and oxygen carrier stability, The preparation method is simple and easy, and the effect of being convenient for popularization and application

Active Publication Date: 2021-07-20
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] As an important means of life-saving, clinical blood transfusion is widely used in clinical operations, disaster relief and war wound treatment, but there are still many problems in blood transfusion: 1. Blood may carry pathogens (hepatitis virus, HIV, etc.), and some diseases may pass through Transfusion transmission; 2. Blood type mismatch leads to agglutination effect, which can threaten the life of the transfused person; 3. Shortage of blood sources; 4. The short storage time of blood limits its treatment of acute and critically ill patients and in emergency situations application under
At present, a variety of artificial oxygen carrier products have been used clinically as blood substitutes. These artificial oxygen carriers include perfluorocarbons, polyhemoglobin, cross-linked hemoglobin, hemoglobin liposomes, etc., but perfluorocarbons need to be refrigerated. , must inhale high oxygen with a concentration of more than 95% at the same time during infusion, and the biological half-life is relatively short, and there are serious side effects
Polyhemoglobin and cross-linked hemoglobin, because the hemoglobin molecule is not protected by other molecules, it is difficult to maintain the functional stability of hemoglobin in the body, and it is easy to cause nephrotoxicity
Hemoglobin liposomes have poor structural stability in the body, and the encapsulated hemoglobin is easy to leak

Method used

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  • Hybrid protein nano oxygen carrier and its preparation method and application
  • Hybrid protein nano oxygen carrier and its preparation method and application
  • Hybrid protein nano oxygen carrier and its preparation method and application

Examples

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preparation example Construction

[0037] According to the second aspect of the present invention, the present invention provides a method for preparing the above hybrid protein nano oxygen carrier, comprising the following steps:

[0038] (a) mixing the reducing agent with serum albumin to perform a reduction reaction to obtain reduced serum albumin;

[0039] (b) Homogenously mixing reduced serum albumin and hemoglobin to prepare a hybrid protein nano-oxygen carrier.

[0040] The preparation method of the hybrid protein nano-oxygen carrier provided by the invention is simple and easy, easy to operate and popularize, and suitable for large-scale preparation.

[0041] In step (a), the reduction reaction between the reducing agent and the serum albumin breaks the disulfide bond inside the serum albumin molecule to obtain the reduced serum albumin molecule in a random state. In step (b), the reduced serum albumin molecule cross-links with the free sulfhydryl group of hemoglobin, reconstitutes an intermolecular di...

Embodiment 1

[0068] This example provides a hybrid protein nano-oxygen carrier, which is prepared according to the following steps:

[0069] (a) Co-dissolve 300mg bovine serum albumin and 30mg dithiothreitol in 5mL deionized water, shake at room temperature for 1 hour, pour the solution into a dialysis bag (3kD), and dialyze in anaerobic deionized water for 12 hours to obtain Reduced serum albumin solution, dilute the reduced serum albumin solution to 6 mL with deionized water to obtain a reduced serum albumin solution with a concentration of 50 mg / mL;

[0070] (b) Dissolve 20 mg of reduced serum albumin solution and 10 mg of hemoglobin in 2 mL of pure water and homogenize for 10 minutes, adjust the pH value to 7 with sodium bicarbonate, and drop into the mixed solution at a rate of 1 mL / min under strong stirring Add 2 mL of absolute ethanol, stir and homogenize for 0.5 hours, put the mixed solution into a dialysis bag with a molecular weight cut-off of 100 kD, and dialyze for 12 hours to ...

Embodiment 2

[0072] This example provides a hybrid protein nano-oxygen carrier, which is prepared according to the following steps:

[0073] (a) Dissolve 300mg of human serum albumin and 300mg of homocysteine ​​in 5mL of deionized water, shake at room temperature for 1 hour, pour the solution into a dialysis bag (3kD), and dialyze in anaerobic deionized water for 12 hours to obtain Reduced serum albumin solution, dilute the reduced serum albumin solution to 6 mL with deionized water to obtain a reduced serum albumin solution with a concentration of 50 mg / mL;

[0074] (b) Dissolve 20 mg of reduced serum albumin solution and 0.4 mg of hemoglobin in 2 mL of pure water and homogenize for 180 minutes, adjust the pH value to 9 with sodium bicarbonate, and add to the mixed solution at a speed of 1 mL / min under vigorous stirring. Add 5 mL of absolute ethanol dropwise, stir and homogenize for 12 hours, put the solution into a dialysis bag with a molecular weight cut-off of 300 kD, and dialyze for 1...

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Abstract

The invention provides a hybrid protein nano-oxygen carrier and its preparation method and application, which relates to the technical field of oxygen carrier, and is mainly composed of hemoglobin and serum albumin covalently bonded through disulfide bonds, and its size is 20-200 nanometers, which relieves The existing artificial oxygen carrier has poor stability in the body and may have technical problems of side effects. Through the covalent combination of serum albumin and hemoglobin through a disulfide bond, the hemoglobin is protected and has good biocompatibility and oxygen-carrying properties in the body. Stability, can efficiently transport oxygen to the body, and will not cause cross-infection and agglutination reaction, and has no technical effect of side effects. In addition, the hybrid protein nano-oxygen carrier provided by the invention has a stable structure, can be synthesized on a large scale, and has a storage time of more than two years, which is convenient for popularization and application.

Description

technical field [0001] The invention relates to the technical field of oxygen carriers, in particular to a hybrid protein nano-oxygen carrier and its preparation method and application. Background technique [0002] As an important means of life-saving, clinical blood transfusion is widely used in clinical operations, disaster relief and war wound treatment, but there are still many problems in blood transfusion: 1. Blood may carry pathogens (hepatitis virus, HIV, etc.), and some diseases may pass through Transfusion transmission; 2. Blood type mismatch leads to agglutination effect, which can threaten the life of the transfused person; 3. Shortage of blood sources; 4. The short storage time of blood limits its treatment of acute and critically ill patients and in emergency situations under the application. [0003] In recent years, nanotechnology has enabled the rapid development of artificial oxygen carriers, which has attracted widespread attention. Artificial oxygen car...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/64A61K38/42C07K19/00A61P7/00
CPCA61K38/42C07K14/765C07K14/805C07K19/00
Inventor 蔡林涛罗震宇郑明彬陈志宽田浩陈泽
Owner SHENZHEN INST OF ADVANCED TECH
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