Advances in Proximity-Assisted Bioconjugation

IF 17.7 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Mary Canzano, Gonçalo J. L. Bernardes
{"title":"Advances in Proximity-Assisted Bioconjugation","authors":"Mary Canzano, Gonçalo J. L. Bernardes","doi":"10.1021/acs.accounts.5c00368","DOIUrl":null,"url":null,"abstract":"Proximity-induced chemistry (PIC) refers to the transient reactivity between two or more molecules upon physical closeness which are otherwise unreactive. Harnessed by nature to control fundamental biological processes such as transcription and signal transduction, PIC increases the probability of correctly oriented, effective collisions, facilitating fundamental cellular processes. Within the field of chemical biology, PIC has been employed for several clinically relevant purposes, including the degradation of aberrant biomolecules and construction of protein therapeutics. This Account focuses on the application of PIC strategies for the development of site-specific bioconjugation techniques, termed proximity-assisted bioconjugation (PAB). Site-specific bioconjugation refers to the precise modification of biomolecules to generate homogeneous products. Such techniques are vital for the development of protein therapeutics including antibody–drug conjugates (ADCs), the investigation of the biological mechanisms of post-translational modifications (PTMs), and the visualization of biomolecular interactions <i>in vitro</i> and <i>in vivo</i>. While numerous strategies have been developed, many suffer from poor yields, limited product stability, demanding experimental procedures, and/or a lack of regioselectivity. Thus, PIC principles have been implemented to address these limitations, leading to the development of PAB strategies which achieve precise, regioselective modification of biomolecules. In this Account, we describe the development of PAB techniques within our group at the University of Cambridge and Instituto de Medicina Molecular (iMM) over the past five years. Our journey with PAB began serendipitously while investigating maleic acid derivatives for cysteine bioconjugation. Here, we discovered the secondary participation of proximal lysines on Trastuzumab-V205C and Gemtuzumab-V205C, conjugatable THIOMAB antibodies commonly used in ADCs, leading to the formation of distinct bioconjugate products relative to IgGs without such lysines. Further investigation into the proximal lysine (K207) of Trastuzumab-V205C revealed that residue 207 could be harnessed directly or mutated to precisely tune the stability of ADCs due to proximity interactions between K207 and covalent modifications of C205. Considering that two Trastuzumab drug conjugates are approved for clinical use, these findings have contributed to the evolving understanding of the chemical landscape of this antibody and help inform future ADC design and development. Further, we describe efforts from our group to develop two distinct PAB approaches: regioselective lysine acetylation of histone H3 and phage display-compatible peptide cyclization. These strategies combine induced-proximity with traditional bioconjugation techniques to enable regioselective modification of biomolecules which are historically difficult to selectively modify. These methods are readily adaptable to related systems and serve as representative examples of how to successfully develop PAB strategies for desired applications. In short, this Account highlights our group’s contributions to and insights on PAB methodologies wherein we illustrate how PIC can be thoughtfully applied to bioconjugation techniques for various aims including regioselective bioconjugation and enhanced bioconjugate stability. We expect that PAB approaches will continue to diversify bioconjugation applications and greatly expand the toolkit of chemical biologists.","PeriodicalId":1,"journal":{"name":"Accounts of Chemical Research","volume":"42 1","pages":""},"PeriodicalIF":17.7000,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Accounts of Chemical Research","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1021/acs.accounts.5c00368","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
引用次数: 0

Abstract

Proximity-induced chemistry (PIC) refers to the transient reactivity between two or more molecules upon physical closeness which are otherwise unreactive. Harnessed by nature to control fundamental biological processes such as transcription and signal transduction, PIC increases the probability of correctly oriented, effective collisions, facilitating fundamental cellular processes. Within the field of chemical biology, PIC has been employed for several clinically relevant purposes, including the degradation of aberrant biomolecules and construction of protein therapeutics. This Account focuses on the application of PIC strategies for the development of site-specific bioconjugation techniques, termed proximity-assisted bioconjugation (PAB). Site-specific bioconjugation refers to the precise modification of biomolecules to generate homogeneous products. Such techniques are vital for the development of protein therapeutics including antibody–drug conjugates (ADCs), the investigation of the biological mechanisms of post-translational modifications (PTMs), and the visualization of biomolecular interactions in vitro and in vivo. While numerous strategies have been developed, many suffer from poor yields, limited product stability, demanding experimental procedures, and/or a lack of regioselectivity. Thus, PIC principles have been implemented to address these limitations, leading to the development of PAB strategies which achieve precise, regioselective modification of biomolecules. In this Account, we describe the development of PAB techniques within our group at the University of Cambridge and Instituto de Medicina Molecular (iMM) over the past five years. Our journey with PAB began serendipitously while investigating maleic acid derivatives for cysteine bioconjugation. Here, we discovered the secondary participation of proximal lysines on Trastuzumab-V205C and Gemtuzumab-V205C, conjugatable THIOMAB antibodies commonly used in ADCs, leading to the formation of distinct bioconjugate products relative to IgGs without such lysines. Further investigation into the proximal lysine (K207) of Trastuzumab-V205C revealed that residue 207 could be harnessed directly or mutated to precisely tune the stability of ADCs due to proximity interactions between K207 and covalent modifications of C205. Considering that two Trastuzumab drug conjugates are approved for clinical use, these findings have contributed to the evolving understanding of the chemical landscape of this antibody and help inform future ADC design and development. Further, we describe efforts from our group to develop two distinct PAB approaches: regioselective lysine acetylation of histone H3 and phage display-compatible peptide cyclization. These strategies combine induced-proximity with traditional bioconjugation techniques to enable regioselective modification of biomolecules which are historically difficult to selectively modify. These methods are readily adaptable to related systems and serve as representative examples of how to successfully develop PAB strategies for desired applications. In short, this Account highlights our group’s contributions to and insights on PAB methodologies wherein we illustrate how PIC can be thoughtfully applied to bioconjugation techniques for various aims including regioselective bioconjugation and enhanced bioconjugate stability. We expect that PAB approaches will continue to diversify bioconjugation applications and greatly expand the toolkit of chemical biologists.

Abstract Image

近距离辅助生物偶联研究进展
邻近诱导化学(PIC)是指两个或多个分子之间在物理上接近时的短暂反应性,否则它们是不反应的。利用自然控制转录和信号转导等基本生物过程,PIC增加了正确定向、有效碰撞的可能性,促进了基本的细胞过程。在化学生物学领域,PIC已被用于几种临床相关目的,包括异常生物分子的降解和蛋白质疗法的构建。本报告侧重于PIC策略在位点特异性生物偶联技术开发中的应用,称为邻近辅助生物偶联(PAB)。位点特异性生物偶联是指对生物分子进行精确修饰以产生均质产物。这些技术对于蛋白质疗法的发展至关重要,包括抗体-药物偶联物(adc),研究翻译后修饰(PTMs)的生物学机制,以及体外和体内生物分子相互作用的可视化。虽然已经开发了许多策略,但许多策略都存在产量低,产品稳定性有限,实验程序要求高和/或缺乏区域选择性的问题。因此,PIC原则已被实施,以解决这些限制,导致PAB策略的发展,实现生物分子的精确,区域选择性修饰。在这篇文章中,我们描述了我们在剑桥大学和分子医学研究所(iMM)的团队在过去五年中PAB技术的发展。我们的旅程与PAB偶然开始在研究马来酸衍生物的半胱氨酸生物偶联。在这里,我们发现近端赖氨酸在adc中常用的可偶联THIOMAB抗体Trastuzumab-V205C和Gemtuzumab-V205C上的二次参与,导致相对于不含赖氨酸的igg形成不同的生物偶联产物。对Trastuzumab-V205C的近端赖氨酸(K207)的进一步研究表明,由于K207与C205的共价修饰之间的邻近相互作用,残基207可以直接利用或突变来精确调节adc的稳定性。考虑到两种曲妥珠单抗药物偶联物已被批准用于临床,这些发现有助于不断发展对该抗体化学景观的理解,并有助于为未来ADC的设计和开发提供信息。此外,我们描述了我们小组开发两种不同的PAB方法的努力:组蛋白H3的区域选择性赖氨酸乙酰化和噬菌体显示兼容肽环化。这些策略结合了诱导接近和传统的生物偶联技术,使生物分子的区域选择性修饰成为可能,而这些生物分子在历史上很难被选择性修饰。这些方法很容易适用于相关系统,并作为如何成功地为期望的应用开发PAB策略的代表性示例。简而言之,本报告强调了我们小组对PAB方法的贡献和见解,其中我们说明了PIC如何能够周到地应用于生物偶联技术,以实现各种目的,包括区域选择性生物偶联和增强生物偶联稳定性。我们期望PAB方法将继续使生物偶联应用多样化,并大大扩展化学生物学家的工具箱。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信