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CN-118084918-B - Chemiluminescent probe, preparation method thereof and application thereof in detection of beta amyloid

CN118084918BCN 118084918 BCN118084918 BCN 118084918BCN-118084918-B

Abstract

The invention discloses a chemiluminescent probe, a preparation method thereof and application thereof in detecting beta amyloid. The structural formula of the chemiluminescent probe is shown as formula (I). The chemiluminescent probe prepared in the invention has longer chemiluminescent wavelength, superior beta amyloid binding capacity and blood brain barrier crossing capacity, and can be used for detecting beta amyloid content and early diagnosis of Alzheimer's disease.

Inventors

  • YANG JING
  • BO BO
  • LIU SHUAINAN

Assignees

  • 中国药科大学

Dates

Publication Date
20260508
Application Date
20240129

Claims (11)

  1. 1. A chemiluminescent probe having a structural formula as shown in (I): Wherein R is Or (b) 。
  2. 2. The chemiluminescent probe of claim 1 wherein the chemiluminescent probe has any one of the following chemical structures: 。
  3. 3. a method of preparing a chemiluminescent probe according to any one of claims 1-2 comprising the steps of: (1) Adding compound 1, ((1, 3-dioxolan-2-yl) methyl) triphenyl phosphonium bromide, anhydrous potassium carbonate and 18-crown ether-6 into an organic solvent, extracting and concentrating after the reaction is finished to obtain a crude product, adding the crude product and hydrochloric acid into tetrahydrofuran, and separating and purifying after the reaction is finished to obtain a compound 2; (2) N-butyllithium and 2, 2, 6, 6-tetramethylpiperidine are added into an organic solvent in protective gas nitrogen, bis- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) methane and a compound 2 are added, and after the reaction is finished, the compound 3 is obtained through separation and purification; (3) Adding a compound 3, 5-bromo-2-aminopyrazine and tetraphenylpalladium phosphate into a1, 4-dioxane containing saturated sodium carbonate solution, and separating and purifying after the reaction is finished to obtain a compound 4; (4) Adding the compound 4, methylglyoxal-1, 1-dimethyl acetal, 6M hydrochloric acid and water into an organic solvent, and standing to precipitate after the reaction is finished to obtain the chemiluminescent probe.
  4. 4. The method of preparing a chemiluminescent probe of claim 3 wherein in step (1), the molar ratio of compound 1, ((1, 3-dioxolan-2-yl) methyl) triphenylphosphine bromide, anhydrous potassium carbonate, and 18-crown-6 is 1:3:4:0.01 to 0.05.
  5. 5. The method of preparing a chemiluminescent probe of claim 3 wherein in step (2) the molar ratio of compound 2, bis- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) methane, n-butyllithium, and 2, 2, 6, 6-tetramethylpiperidine is 1:1.5 to 2:2:3.
  6. 6. The method for preparing a chemiluminescent probe of claim 3 wherein in step (3), the molar ratio of the compound 3, 5-bromo-2-aminopyrazine to tetraphenylpalladium is 1:1.5-2:0.01-0.05.
  7. 7. The method of preparing a chemiluminescent probe of claim 3 wherein in step (4), the molar ratio of compound 4 to methylglyoxal-1, 1-dicarboxaldehyde is 1:1.3-1.8.
  8. 8. Use of a chemiluminescent probe according to any one of claims 1-2 in the preparation of a formulation for detecting amyloid beta.
  9. 9. Use of a chemiluminescent probe according to any one of claims 1-2 for the detection of amyloid beta, which is not targeted for the treatment and/or diagnosis of a disease.
  10. 10. The use according to claim 9, wherein a chemiluminescent probe is added to a test solution comprising beta amyloid protein to obtain a test solution, and wherein the beta amyloid protein in the test solution is detected qualitatively or quantitatively.
  11. 11. The use according to claim 10, wherein the chemiluminescent probe emits in the wavelength range of 570-620 nm without an excitation light source.

Description

Chemiluminescent probe, preparation method thereof and application thereof in detection of beta amyloid Technical Field The invention relates to a chemiluminescent probe and a preparation method and application thereof, in particular to a chemiluminescent probe and a preparation method thereof and application thereof in detecting beta amyloid, belonging to the field of biological medicine. Background Alzheimer's Disease (AD) is a latent neurodegenerative Disease, clinically characterized by global dementia such as dysmnesia, aphasia, disuse, disbelief, and personality and behavior changes, which severely affects human health and quality of life. Unfortunately, there is currently no therapeutic regimen that can effectively prevent and reverse the course of AD, and methods that can diagnose AD early are particularly important. The pathogenesis of AD is currently unknown, and among the many hypotheses of the etiology of AD, the cascade of amyloid beta (AmyLoid beta, abeta) has been widely accepted, namely, the cleavage of Amyloid Precursor Protein (APP) by enzymes to form Abeta monomers, the aggregation of Abeta monomers to form plaques, the interference of synaptic transmission of neuronal signals, and finally the death of neuronal cells. A large number of researches show that the deposition of Abeta protein starts to occur 10-20 years before the AD patient has symptoms, so that the sensitive detection of Abeta is expected to realize the early diagnosis of AD. The optical probe has higher space-time resolution capability and is widely used for in-vivo and in-vitro detection research of Abeta, but most of the current optical probes are fluorescent probes, and the signal to noise ratio of imaging is low due to the requirement of external high-energy excitation light. The chemiluminescence is based on chemical reaction to generate light, and an external light source is not needed, so that background signals brought by excitation light can be effectively reduced, and the detection sensitivity is higher. However, the emission wavelength of the chemiluminescent probes reported for detecting aβ is much shorter and the ability to penetrate the blood brain barrier is poor. Disclosure of Invention The invention aims to provide a chemiluminescent probe, a preparation method of the chemiluminescent probe and an application of the chemiluminescent probe in detecting beta amyloid. The technical scheme is that the chemiluminescent probe has a structural formula shown in (I): (I) Wherein R is 、、Or (b)。 Preferably, the chemiluminescent probe has any one of the following chemical structures: in another aspect, the present invention provides a method for preparing the chemiluminescent probe, comprising the steps of: (1) Adding compound 1, ((1, 3-dioxolan-2-yl) methyl) triphenyl phosphonium bromide, anhydrous potassium carbonate and 18-crown ether-6 into an organic solvent, extracting and concentrating after the reaction is finished to obtain a crude product, adding the crude product and hydrochloric acid into tetrahydrofuran, and separating and purifying after the reaction is finished to obtain a compound 2; (2) N-butyllithium and 2, 2, 6, 6-tetramethylpiperidine are added into an organic solvent in protective gas nitrogen, bis- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) methane and a compound 2 are added, and after the reaction is finished, the compound 3 is obtained through separation and purification; (3) Adding a compound 3, 5-bromo-2-aminopyrazine and tetraphenylpalladium phosphate into a1, 4-dioxane containing saturated sodium carbonate solution, and separating and purifying after the reaction is finished to obtain a compound 4; (4) Adding the compound 4, methylglyoxal-1, 1-dimethyl acetal, 6M hydrochloric acid and water into an organic solvent, and standing to precipitate after the reaction is finished to obtain the chemiluminescent probe. Preferably, in the step (1), the molar ratio of the compound 1, ((1, 3-dioxolan-2-yl) methyl) triphenyl phosphonium bromide, anhydrous potassium carbonate and 18-crown ether-6 is 1:3:4:0.01-0.05. Preferably, in step (1), the organic solvent is anhydrous toluene. Preferably, in the step (1), the dosage of the anhydrous toluene is calculated by 5mL of anhydrous toluene in a ratio of 4-diethylaminobenzaldehyde per millimole. In the step (1), the use amount of the hydrochloric acid solution is calculated according to the proportion of 0.5mL of hydrochloric acid in each millimole of 4-diethylaminobenzaldehyde. Preferably, in step (1), the protective gas is nitrogen or argon. Preferably, in the step (1), the reaction time is 10-12 hours. Preferably, in step (1), the extraction is performed with dichloromethane. Preferably, in the step (1), the separation and purification are performed by silica gel column chromatography. The silica gel column chromatography is to perform chromatographic separation in a 100-200 mesh silica gel column, and adopts a mixed solvent of petroleum ether and e