Fluorescent probe for detecting pyroglutamamide aminopeptidase I as well as preparation method and application of fluorescent probe

A technology of pyroglutamylamino and fluorescent probes, applied in the field of fluorescent probes for detecting pyroglutamyl aminopeptidase I and its preparation, achieving the effects of excellent photophysical properties, low cost, and strong penetration

Active Publication Date: 2021-12-28
西安天工生物医药研究所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, two-photon fluorescent probes that can specifically detect PGP-1 in clinical human samples have not yet been reported, and it is urgent to explore

Method used

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  • Fluorescent probe for detecting pyroglutamamide aminopeptidase I as well as preparation method and application of fluorescent probe
  • Fluorescent probe for detecting pyroglutamamide aminopeptidase I as well as preparation method and application of fluorescent probe
  • Fluorescent probe for detecting pyroglutamamide aminopeptidase I as well as preparation method and application of fluorescent probe

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preparation example Construction

[0036] A kind of synthetic method for detecting the fluorescent probe of pyroglutamyl aminopeptidase I, comprises the following steps:

[0037] Mix LXY, HATU and DIPEA, completely dissolve in dry dichloromethane solution, stir and react at 0°C; then add TPAN solution, further stir and react at room temperature, and then purify to obtain a deep red solid BH1-BOC;

[0038] Dissolve BH1-BOC in anhydrous dichloromethane, add the prepared trifluoroacetic acid solution under the condition of ice-salt bath, and carry out stirring reaction at room temperature after the dropwise addition; spin to dry the solvent, then add dichloromethane to it, repeat Several times, until all the trifluoroacetic acid was spin-dried; the crude product was purified by TM to obtain a red solid BH1.

[0039] The ability of the probe to analyze and detect PGP-1 was evaluated in vitro; with 350nm as excitation, after adding PGP-1, the fluorescence spectrum of the probe will change obviously. Probe BH1 exhib...

Embodiment 1

[0054] 1. Synthesis of Dimethyl(R)-6-(1-(tert-butoxycarbonyl)-5-oxopyrrolidine-2-carboxamido)naphthalene-2,3-dicarboxylate(BH1-BOC)

[0055] Add dimethyl 6-aminonaphthalene-2,3-dicarboxylate (LXY; 115mg, 0.5mmol), 2-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluroniumhexafluorophosphate (HATU; 190mg, 0.5mmol) and N,N-Diisopropylethylamine (DIPEA; 200μL, 1.25mmol), which were completely dissolved in dry dichloromethane solution (25mL), and the above reactants were stirred at 0°C for 30 minutes. Then, post-TPAN (130 mg, 0.5 mmol) dissolved in dichloromethane (5 mL) was further added, followed by further stirring at room temperature for 4 hours. After the reaction was over, distilled under reduced pressure and spin-dried the solvent, the crude product was purified by silica gel chromatography, using petroleum ether / ethyl acetate (15 / 1, v / v) as eluent to obtain a dark red solid BH1-BOC ( 70 mg), yield: 14%. BH1-BOC: 1 H NMR (500MHz, DMSO-d 6 )δppm: 10.80(s, 1H), 8.44(s, 1H), ...

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Abstract

The invention discloses a fluorescent probe for detecting pyroglutamamide aminopeptidase I as well as a preparation method and application thereof. The method comprises the following steps: mixing LXY, HATU and DIPEA, completely dissolving the mixture in a dry dichloromethane solution, and carrying out a stirring reaction at 0 DEG C; then adding a TPAN solution, further conducting stirring and reacting at room temperature, and carrying out purifying after the reaction is finished so as to obtain a dark red solid BH1-BOC; dissolving BH1-BOC in anhydrous dichloromethane, adding a prepared trifluoroacetic acid solution under an ice salt bath condition, and carrying out room-temperature stirring reaction after dropwise adding is completed; spin-drying a solvent, adding dichloromethane, and repeating the above operations multiple times until the trifluoroacetic acid is completely spin-dried; and purifying a crude product to obtain a red solid BH1.

Description

technical field [0001] The invention belongs to the field of analysis and detection of small molecule fluorescent probes, in particular to a fluorescent probe for detecting pyroglutamyl aminopeptidase I (PGP-1), a preparation method and application thereof. Background technique [0002] Pyroglutamyl aminopeptidase I (PGP-1, EC 3.4.19.3) is a typical cytoplasmic cysteine ​​peptidase. PGP-1 is widely distributed in bacteria, plants and animal tissues, and can be obtained from endogenous proteins and bioactive peptides (vasopressin, luteinizing hormone-releasing hormone, gastrin, thyrotropin-releasing hormone and immunoglobulin , etc.) specifically hydrolyzes L-pyroglutamic acid (L-pGlu) residues at the amino terminus. Thus, PGP-1 has a pronounced effect on determining the amino-terminal sequences of N-blocked peptides and proteins containing L-pGlu residues. The enzyme-catalyzed product L-pGlu also has a variety of pharmacological properties, mainly including excitatory amin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D207/22C09K11/06G01N21/64
CPCC07D207/22C09K11/06G01N21/6428C09K2211/1029Y02P20/55
Inventor 柏桦方斌李林彭勃胡文博姚传好于海东傅莉黄维
Owner 西安天工生物医药研究所有限公司
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