Chinese Journal of Organic Chemistry ›› 2024, Vol. 44 ›› Issue (7): 2257-2264.DOI: 10.6023/cjoc202312029 Previous Articles     Next Articles

ARTICLES

新型查尔酮衍生物氨肽酶N荧光探针的设计及应用

胡亦然a,b, 张巳亮a,b, 罗海艳a,b, 赵璐瑶a,b, 郭旭东a,b, 王双青a, 胡睿a,*(), 杨国强a,b,*()   

  1. a 中国科学院化学研究所光化学重点实验室 北京分子科学国家研究中心 北京 100190
    b 中国科学院大学 北京 100039
  • 收稿日期:2023-12-30 修回日期:2024-03-22 发布日期:2024-04-10
  • 基金资助:
    国家自然科学基金(22273110); 中国科学院青年创新促进会(2020035)

Design and Application of a Novel Chalcone Derivative Fluorescent Probe for Aminopeptidase N

Yiran Hua,b, Siliang Zhanga,b, Haiyan Luoa,b, Luyao Zhaoa,b, Xudong Guoa,b, Shuangqing Wanga, Rui Hua(), Guoqiang Yanga,b()   

  1. a Beijing National Laboratory for Molecular Sciences, Key Laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190
    b University of Chinese Academy of Sciences, Beijing 100039
  • Received:2023-12-30 Revised:2024-03-22 Published:2024-04-10
  • Contact: E-mail: hurui@iccas.ac.cn;gqyang@iccas.ac.cn
  • Supported by:
    National Natural Science Foundation of China(22273110); Youth Innovation Promotion Association of Chinese Academy of Sciences(2020035)

Aminopeptidase N (APN) is a widely expressed hydrolytic enzyme in organisms that can hydrolyze neutral or alkaline amino acids at the N-terminus of protein polypeptide chains. Overexpressed APN may indicate disorders of biosystems, such as various tumors, drug-induced liver injury (DILI), and glomerulonephritis, and is widely used for early diagnosis and therapy of related diseases. Particularly, elevated APN level in urine is becoming one of the key markers of early kidney injury due to its special sensitivity to glomerulonephritis than other proteins. Herein, based on the structure of 4-amino chalcone, a novel fluorescent probe Ala-OMN for detecting APN was designed and synthesized by caging the 4-amino of OMN by alanine. Ala-OMN is non-fluorescent in phosphate buffered saline (PBS). After reaction with APN, the caging group alanine was hydrolyzed and OMN was released, which further combined with bovine serum protein (BSA) to form a 1∶1 complex, concomitantly with a bright green fluorescence signal for indicating APN. Ala-OMN exhibits excellent sensitivity, selectivity, and anti-interference ability in response to APN, with a detection limit as low as 0.058 ng/mL. Ala-OMN has been successfully applied to detect APN concentration in urine samples, and the results are in good agreement with commercial reagent kits, suggesting that Ala-OMN could be a new diagnostic reagent for the clinical detection of early glomerulonephritis.

Key words: chalcone derivatives, aminopeptidase N (APN), fluorescent probe, bovine serum protein (BSA)