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Chemiluminescent probe for detecting fibroblast activating protein as well as synthesis method and application of chemiluminescent probe

A fibroblast, chemiluminescence technology, applied in chemical instruments and methods, chemiluminescence/bioluminescence, analysis by chemical reaction of materials, etc., to achieve important application value and expand the scope of application.

Active Publication Date: 2021-06-25
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection methods of FAPα mainly include RT-PCR, immunohistochemical method, Western bolt method, fluorescence method, etc., and there is a gap in the technology of chemiluminescence method for the determination of FAPα.

Method used

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  • Chemiluminescent probe for detecting fibroblast activating protein as well as synthesis method and application of chemiluminescent probe
  • Chemiluminescent probe for detecting fibroblast activating protein as well as synthesis method and application of chemiluminescent probe
  • Chemiluminescent probe for detecting fibroblast activating protein as well as synthesis method and application of chemiluminescent probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Synthesis of the adamantane-dioxetane chemiluminescent probe CFCL for the detection of fibroblast activation protein (see the attached figure 1 shown).

[0042] (1) The synthesis of compound 7, the specific steps are:

[0043] (1) Dissolve m-hydroxybenzaldehyde (25.6 g, 209.6 mmol, 1 equiv.) in 150 mL of 90 % acetic acid, and after the temperature of the reaction solution drops to 0°C, add tert-butyl hypochlorite (25.0 g, 230.6 mmol, 1.1 equiv.); HPLC monitors the reaction; after the reaction is complete, filter to obtain the product (recorded as compound 1), as a white solid 13.8g (42.2 % crude product yield); MS (ESI-): m / C 7 h 5 ClO 2 Theoretical values: 155.00, 156.99; Visible [M-H]-154.88, 156.88;

[0044] (2) Dissolve compound 1 (100 %, 10.8 g, 69.2 mmol, 1 equiv.) in 100 mL of methanol, add tetrabutylammonium tribromide (1.7 g, 3.5 mmol, 0.05 equiv.) and Trimethyl orthoformate (11.8 g, 110.8 mmol, 1.6 equiv.), reacted at room temperature for 4 ...

Embodiment 2

[0057] Example 2: Synthesis of an adamantane-dioxetane chemiluminescence probe BFCL for detecting fibroblast activation protein. (For the synthetic route see figure 2 shown), the specific steps are:

[0058] (1) Combine tert-butoxycarbonyl-glycine-proline-OH (1.1 g, 44.04 mmol, 1 equiv.) and ethyl 2-ethoxy-1(2H)-quinolinecarboxylate (2.0 g, 8.08 mmol , 2 equiv.) was dissolved in 20 mL of dichloromethane, and a dichloromethane solution of p-aminobenzyl alcohol (1.0 g, 8.08 mmol, 2 equiv.) was added dropwise. The reaction was monitored by high performance liquid chromatography. After the reaction, add appropriate amount of dichloromethane to dilute and wash with water. The organic phases were combined, dried with saturated sodium sulfate solid, the solvent was removed by rotary evaporation in vacuum, and purified by silica gel column chromatography. The obtained product (referred to as compound b1) was a white solid, 1.24 g (82.7 % yield);

[0059] (2) Compound b1 (50 mg, 0...

Embodiment 3

[0061] Example 3: CFCL or BFCL Determination of Fibroblast Activation Protein in Vitro.

[0062] Prepare a series of FAPα solutions with different concentrations, take 100 μL of each concentration into small wells of a black 96-well plate, add 100 μL of 40 μM CFCL or BFCL chemiluminescent probes to each well, and use Xenogen IVIS ® The in vivo imager takes pictures and performs quantitative analysis by the instrument's own software. Such as image 3 As shown, the response value increased with the increase of FAPα concentration, and there was a good linear relationship between the response value and the FAPα concentration.

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Abstract

The invention belongs to the technical field of small-molecule chemiluminescence probes, and particularly relates to a chemiluminescence probe for detecting fibroblast activation protein (FAP[alpha]) as well as a synthesis method and application of the chemiluminescence probe. The chemiluminescent probe disclosed by the invention is an adamantane-dioxetane chemiluminescent probe and comprises two types, namely CFCL and BFCL. An adamantane-dioxetane derivative and an FAP[alpha] sensitive group are subjected to a high performance liquid chromatography monitoring reaction in an anhydrous inert solvent in the presence of an acid-binding agent and a catalyst, and the adamantane-dioxetane chemiluminescent probe is prepared. The chemiluminescent probe can be used for in-vivo and in-vitro detection of FAP[alpha], including determination of FAP[alpha] in cells, tissues or plasma, and imaging of FAP[alpha] in animal bodies. The chemiluminescent probe can also be used for evaluating and screening FAP[alpha] inhibitors. According to the present invention, the adamantane-dioxetane is improved to obtain the chemiluminescence probe, such that the FAP[alpha] detection is achieved, the important technical breakthrough is provided, and the application range of the chemiluminescence probe is further expanded.

Description

technical field [0001] The invention belongs to the technical field of small molecule chemiluminescent probes, and in particular relates to a chemiluminescent probe for detecting fibroblast activation protein, a synthesis method and application thereof. Background technique [0002] Compared with fluorescence technology, chemiluminescence originates from chemical reactions, does not require external excitation light sources, and has the unique advantage of not being interfered by light scattering and autofluorescence; compared with bioluminescence, chemiluminescence does not need to rely on various luciferase-substrates. Specific reactions to substances. Chemiluminescent systems, including luminol, peroxalate, etc., have complex composition systems, weak applicability in physiological environments, and the participation of oxidative substances in the reaction, and the application of optical analysis in vivo is mostly limited to Determination of hydrogen peroxide, active oxy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D405/12C09K11/07G01N21/76A61K49/00
CPCC07D405/12C09K11/07G01N21/76A61K49/0013
Inventor 傅安辰曹志娟王宏博毛彦佳孙茹
Owner FUDAN UNIV
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