Method for detecting and accelerating beta-amyloid protein aggregation by means of lanthanide fluorescence complex
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A complex and fluorescent technology, applied in the field of medicine, can solve the problems of detection, acceleration of Aβ aggregation, etc.
Active Publication Date: 2016-09-28
NANJING UNIV OF TECH
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However, it has not been found that it is used to detect and accelerate Aβ aggregation and to develop AD diagnostics.
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Embodiment 1
[0032] Example 1: Detection of Aβ aggregates by lanthanide fluorescent complexes:
[0033] Step 1: Synthesis of the preferred lanthanide fluorescent complex Eu-L, the synthesis route is shown below.
[0034]
[0035] Weigh 2-aminopyridine (0.376g, 4mmol) and α-bromo-p-nitroacetophenone (0.486g, 2mmol), dissolve in 20mL of acetonitrile solution, stir, and reflux at 80°C for 3h, a large amount of red blood appears in the solution The orange-yellow precipitate was suction-filtered, washed with acetonitrile, collected, and dried to obtain an orange-yellow solid (0.520 g, yield: 90%). 1 H NMR (CDCl 3 , 500MHz, δ, ppm): 6.90 (t, 1H, imidazo[1, 2, a]pyridine), 7.30 (t, 1H, imidazo[1, 2, a]pyridine), 7.73 (d, 1H, imidazo[ 1, 2, a] pyridine), 8.01 (s, 1H, imidazo [1, 2, a] pyridine), 8.15 (d, 2H, phenyl), 8.18 (d, 1H, imidazo [1, 2, a] pyridine ), 8.30 (d, 2H, phenyl). ESI-MS found (calcd) for C 13 h 9 N 3 o 2 (m / z): 239.07(239.23)[M+H] + .
[0036]Weigh compound 1 (0.120g...
Embodiment 2
[0046] Example 2: Lanthanide Fluorescent Complex Accelerates Aβ Aggregation:
[0047] Step 1: Synthesis of the preferred lanthanide fluorescent complex Eu-L, the synthesis method is the same as that of Step 1 of Example 1.
[0048] Step 2: Different concentrations of Eu-L (0, 5, 20 and 40 μM) were incubated with Aβ monomer samples in a buffer solution at 37° C. for 7 days to accelerate Aβ aggregation.
[0049] Step 3: Using Eu-L time-resolved fluorescence detection method to track and monitor the acceleration of Aβ aggregation.
[0050] Figure 6 For example two different concentrations of Eu-L to Aβ 40 Accelerated turbidity of aggregation (A 405 ) detection diagram (A) and Thioflavin T (ThT) fluorescence detection diagram (B). The experimental results showed that Aβ 40 Self-aggregation is mainly in the stagnation period (nucleation period) of aggregation in the first 4 days, and enters the elongation period after 4 days. After co-incubation with Eu-L (final concentratio...
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Abstract
The invention discloses a method for detecting and accelerating beta-amyloid protein aggregation by means of a lanthanide fluorescence complex, and belongs to the technical field of medicines. According to the method, the lanthanide fluorescence complex is used for detecting A-beta aggregate, an imidazo[1,2,a]pyridine derivative serves as an antenna group to be coupled with a europium (III)-DTPA luminescence center to prepare a fluorescence complex Eu-L, the antenna group can be selectively combined with A-beta hydrophobic region amino acid residues with the beta-folding conformation, photoinduced electron transfer of amino acid hinders energy transfer from the antenna group to europium (III), Eu-L fluorescence is reduced, and the A-beta aggregate is detected by means of fluorescence changes. According to the method, the lanthanide fluorescence complex is further used for accelerating A-beta aggregation, on the basis of high affinity of the antenna ligand and the A-beta aggregate, the Eu-L can accelerate conversion of A-beta oligomers into corpus fibrosum, the A-beta aggregation process is simultaneously monitored by means of autofluorescence, and a good application prospect in the aspect of AD diagnosis and treatment is achieved.
Description
technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for detecting and accelerating the aggregation of beta amyloid protein with a lanthanide fluorescent complex. Background technique [0002] Alzheimer's disease (AD) is a degenerative disease of the central nervous system with progressive cognitive decline that seriously endangers the physical and mental health of middle-aged and elderly people. The main pathological features of AD are senile plaques (SP) formed by the deposition of β-amyloid protein (Aβ), neurofibrillary tangles (NFTs) formed by the aggregation of abnormally phosphorylated Tau protein, and the loss of neurons in the cerebral cortex and hippocampus. Degeneration or death, in which Aβ plays an important role in the formation of various characteristic pathologies, is the initiating factor and central link in the development of AD. Aβ mainly includes Aβ40 and Aβ42, which are protein fragments p...
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