A kind of serum albumin with metal chelating function, its preparation method and its application in inhibiting the aggregation of β-amyloid protein

A serum albumin and amyloid technology, applied in the field of biomedicine, can solve the problems of insufficient Aβ aggregation inhibition effect, adsorption inhibition of Aβ aggregation, limited metal ion binding ability, etc. Gentle and easy effect

Inactive Publication Date: 2020-11-03
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, under physiological conditions, the concentration of HSA in cerebrospinal fluid is low, which is not enough to inhibit Aβ aggregation through simple adsorption, and it has defects such as poor specificity.
In addition, the binding ability of HSA to metal ions is also very limited, and the inhibitory effect on Aβ aggregation in the presence of metal ions is insufficient.

Method used

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  • A kind of serum albumin with metal chelating function, its preparation method and its application in inhibiting the aggregation of β-amyloid protein
  • A kind of serum albumin with metal chelating function, its preparation method and its application in inhibiting the aggregation of β-amyloid protein
  • A kind of serum albumin with metal chelating function, its preparation method and its application in inhibiting the aggregation of β-amyloid protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Synthesis and characterization of metal chelating functional serum albumin with a molar ratio of iminodiacetic acid to serum albumin of 5

[0043] Weigh 20 mg of serum albumin (HSA), dissolve it in 20 mL of boric acid-borax buffer (20 mmol / L, pH 8.0) to obtain a serum albumin solution with a concentration of 1 mg / mL, and add 55 μL of 1,4-butanediol diglycidyl ether. The above mixture was stirred and reacted at 120rpm and 25°C for 8h. After the reaction, the free 1,4-butanediol diglycidyl ether in the system was removed through a SephadexG-25 gel filtration column, and the mobile phase was boric acid-borax buffer (20mmol / L, pH 8.0). Weigh 0.2 mg of iminodiacetic acid powder and add it to the solution obtained in the previous step. The above mixture was stirred and reacted at 120 rpm and 25° C. for 24 h. After the reaction, the free iminodiacetic acid in the system was removed through a SephadexG-25 gel filtration chromatographic column, and the mobile phase...

Embodiment 2

[0049] Example 2: Synthesis and characterization of metal chelating functional serum albumin with a molar ratio of iminodiacetic acid to serum albumin of 50

[0050] 200 mg of serum albumin was weighed and dissolved in 20 mL of boric acid-borax buffer (20 mmol / L, pH 8.5) to obtain a serum albumin solution with a concentration of 10 mg / mL. , and add 825 μL of 1,4-butanediol diglycidyl ether. The above mixture was stirred and reacted at 120rpm and 37°C for 10h. After the reaction, the free 1,4-butanediol diglycidyl ether in the system was removed by SephadexG-25 gel filtration chromatography column, and the mobile phase was boric acid-borax buffer (20mmol / L, pH 8.5). Weigh 19.8 mg of iminodiacetic acid powder and add it to the solution obtained in the previous step. The above mixture was stirred and reacted at 120 rpm and 37° C. for 36 h. After the reaction, remove free iminodiacetic acid in the system through a SephadexG-25 gel filtration chromatographic column, the mobile p...

Embodiment 3

[0056] Example 3: Synthesis and characterization of acidified serum albumin with a molar ratio of iminodiacetic acid to serum albumin of 400

[0057] Weigh 400 mg of serum albumin, dissolve it in 20 mL of boric acid-borax buffer (20 mmol / L, pH 9.0) to obtain a serum albumin solution with a concentration of 20 mg / mL, and add 2200 μL of 1,4-butanediol disulfide glyceryl ether. The above mixture was stirred and reacted at 120rpm and 50°C for 12h. After the reaction, the free 1,4-butanediol diglycidyl ether in the system was removed by SephadexG-25 gel filtration chromatography column, and the mobile phase was boric acid-borax buffer solution (20mmol / L, pH 9.0). Weigh 316.8 mg of iminodiacetic acid powder and add it to the solution obtained in the previous step. The above mixture was stirred and reacted at 120rpm and 50°C for 48h. After the reaction, the free iminodiacetic acid in the system was removed by SephadexG-25 gel filtration chromatographic column, the mobile phase was w...

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Abstract

The invention relates to a serum albumin with a metal chelating function as well as a preparation method and an application in inhibition for the aggregation of beta-amyloid proteins. The serum albumin with the metal chelating function is obtained through modifying 1,4-butanediol diglycidyl ether on an amino group on the surface of a serum albumin at first, and then modifying a metal chelating agent iminodiacetic acid on an epoxy group of 1,4-butanediol diglycidyl ether, and the average modification number of iminodiacetic acid on each serum albumin ranges from 2 to 40. According to the serum albumin with the metal chelating function, which is disclosed by the invention, the chelating capacity of the serum albumin for metal ions is improved, and then the self-aggregation of the beta-amyloid proteins and an aggregation process of the beta-amyloid proteins under the induction of the metal ions are inhibited, and the cytotoxicity of beta-amyloid protein aggregates is reduced. The serum albumin with the metal chelating function is used for a medicine for treating the diseases related to beta-amyloid protein aggregation. The medicine is used for treating and slowing down Alzheimer disease and other diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a serum albumin with metal chelating function, a preparation method and an application in inhibiting beta-amyloid aggregation. Background technique [0002] Alzheimer's disease (AD) is the most important form of senile dementia, accounting for 50%-70% of all dementias, and is currently a neurodegenerative disease that has attracted much attention (Nature 2014,507,448- 454). The incidence of AD will continue to increase with the increase of human age. As the aging society continues to intensify, it is estimated that by 2050, AD patients will reach 115 million (ACS chemical neuroscience 2014,5,718-730), becoming a major threat to the elderly. one of the diseases. Since there is no effective method to treat AD clinically (Cell 2005, 120, 545-555), as a medical and social problem that seriously affects human health, the prevention and treatment of AD has become a re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765C07K1/113A61K38/38A61K47/54A61P25/28
CPCA61K38/385C07K14/765
Inventor 孙彦谢宝龙余林玲董晓燕
Owner TIANJIN UNIV
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