利用自旋标记肽偶联苯乙烯-马来酸共聚物开发基于esr的基质金属蛋白酶活性诊断探针

IF 1.7 4区 医学 Q3 PHARMACOLOGY & PHARMACY
Masato Sasaki, Kyouka Kato, Yuhei Ohta, Haruka Nakamura, Akihiko Kuniyasu, Shoko Okazaki, Keizo Takeshita
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引用次数: 0

摘要

基质金属蛋白酶(Matrix metalloproteinases, MMPs),特别是MMP-2和MMP-9,在肿瘤生长和恶性过程中发挥着重要作用。因此,测量它们在体内的活性可以提高癌症的诊断和治疗。鉴于近几十年来体内ESR技术在医学和制药领域的应用,我们开发了一种基于依赖于硝基自由基旋转相关时间的ESR光谱变化来评估MMP活性的探针。探针是通过MMP底物肽和聚醚胺将硝基自由基偶联到苯乙烯-马来酸共聚物上合成的。探针的ESR信号被与牛血清白蛋白形成复合物而变宽。当将MMP-2或MMP-9添加到复合物中时,尖锐信号的强度随时间显著增加。这种增加被MMP-2/MMP-9的特异性抑制剂完全抑制,并且在含有混乱底物肽的探针中没有发生。MMP-2降解复合物的比常数(kcat/Km)为4.7 × 103 M-1 s-1,与先前报道的用于癌症治疗和诊断的MMP-2底物相比,低1-2个数量级。这种较低的催化效率归因于相对于其他MMP-2底物较高的Michaelis-Menten常数,表明底物结合亲和力降低。尽管需要提高探针的亲和力,但本研究证明了ESR信号在响应MMP-2和MMP-9活性时增加的机制,强调了其在无创体内评估MMP活性的潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of an ESR-Based Diagnostic Probe for Matrix Metalloproteinase Activity Using a Spin-Labeled Peptide-Conjugated Styrene-Maleic Acid Copolymer.

Matrix metalloproteinases (MMPs), specifically MMP-2 and MMP-9, play a significant role in tumor growth and malignancy. Therefore, measuring their activity in vivo could enhance the diagnosis and treatment of cancer. Given the medical and pharmaceutical applications of the in vivo ESR techniques in recent decades, we developed a probe to evaluate MMP activity based on ESR spectral changes that depend on the rotational correlation time of the nitroxyl radical. The probe was synthesized by conjugating the nitroxyl radical to a styrene-maleic acid copolymer via an MMP substrate peptide and polyetheramine. The ESR signal of the probe was broadened by complex formation with bovine serum albumin. When either MMP-2 or MMP-9 was added to the complex, the intensity of the sharp signal increased markedly over time. This increase was completely inhibited by specific inhibitors of MMP-2/MMP-9 and did not occur with a probe containing scrambled substrate peptides. The specific constant (kcat/Km) for degradation of the complex by MMP-2 was 4.7 × 103 M-1 s-1, comparable to or 1-2 orders of magnitude lower than that of previously reported MMP-2 substrates designed for cancer therapy and diagnosis. This lower catalytic efficiency was attributed to a higher Michaelis-Menten constant relative to other MMP-2 substrates, suggesting a reduced substrate binding affinity. Despite the need for improved probe affinity, this study demonstrates a mechanism in which ESR signals increase in response to MMP-2 and MMP-9 activity, highlighting its potential for noninvasive in vivo assessment of MMP activity.

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来源期刊
CiteScore
3.50
自引率
5.00%
发文量
247
审稿时长
2 months
期刊介绍: Biological and Pharmaceutical Bulletin (Biol. Pharm. Bull.) began publication in 1978 as the Journal of Pharmacobio-Dynamics. It covers various biological topics in the pharmaceutical and health sciences. A fourth Society journal, the Journal of Health Science, was merged with Biol. Pharm. Bull. in 2012. The main aim of the Society’s journals is to advance the pharmaceutical sciences with research reports, information exchange, and high-quality discussion. The average review time for articles submitted to the journals is around one month for first decision. The complete texts of all of the Society’s journals can be freely accessed through J-STAGE. The Society’s editorial committee hopes that the content of its journals will be useful to your research, and also invites you to submit your own work to the journals.
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