单击化学启用[89ZR] Zr-Dota doadimumunoconjugation for Theranostic 89zr-imunopet
Click Chemistry Enables [89Zr]Zr-DOTA Radioimmunoconjugation for Theranostic 89Zr-immunoPET
影响因子:3.90000
分区:化学2区 / 生化研究方法2区 有机化学2区 生化与分子生物学3区 化学：综合3区
发表日期:2024 Nov 20
作者:
Ryota Imura, Jaewoong Jang, Atsuko Nakanishi Ozeki, Hiroyuki Takahashi, Hiroyuki Ida, Youichiro Wada, Yoshitaka Kumakura, Nobuyoshi Akimitsu
摘要
有一个预测，在锆89（89zr）免疫 - 普通术发射术（89ZR-ammmmmunopet）中，在89（89ZR）中使用大环螯合剂1,4,7,10-tetraazacyClododecane-1,4,7,7,10-四乙酸（DOTA）可以增强VIVOSONSONIMIMONIMANICE in insobane invivo stormoine in vivo strance。然而，将[89ZR] ZR-DOTA与单克隆抗体（MAB）共轭仍然是一个挑战，因为[89ZR] ZR-DOTA螯合所需的热处理可导致MAB部分的热变性。我们开发了一种基于四嗪（TZ）连接的双功能DOTA衍生物合成[89ZR] ZR-DOTA-MAB的方法2,2'，2英寸 - （10-（1-（4-（1,2,4,5-四唑-3-基）苯基）-3,21,26-Trioxo-6,9,12,12,15,15,15,18-Pentaoxa-29-C arboxy-2,22,25-三氮烷烷烷-29-基）-1,4,7,10-tetraazacyClododecane-1,4,4,7-三甲基）三acic酸（Dotaga-tz）和逆电子需求Diels-Alder（IEDDA）单击化学反应，将反式环绕烯修饰的mAb偶联到[89zr] Zr-Dotaga而不暴露于热量中。 IEDDA衍生的[89ZR] Zr-Dotaga-Trastuzumab的稳定性通过体外，Ex Vivo和对常规脱脂氧胺（DFO）对应物[89ZR] ZR-DFO-DFO-DFO-trastuzumab的体内测试和比较分析证实。使用[89ZR] Zr-Dotaga-Trastuzumab清楚地可见的人类表皮生长因子受体2阳性恶性肿瘤的体内免疫集成像清楚地可见。与[89ZR] ZR-DFO-trastuzumab相比，从[89ZR] Zr-Dotaga-Trastuzumab处观察到更大的肿瘤对比度。这些发现表明，我们的IEDDA连接方法可以是合成[89ZR] ZR-DOTA-MAB的有效手段，并可以增强在DOTA介导的放射免疫疗法中89ZR-免疫膜的疗法潜力。
Abstract
There have been predictions that the use of the macrocyclic chelating agent 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) in zirconium-89 (89Zr) immuno-positron emission tomography (89Zr-immunoPET) could enhance the in vivo stability of 89Zr radioimmunoconjugates. However, conjugating [89Zr]Zr-DOTA to a monoclonal antibody (mAb) remains a challenge as the heat treatment required for [89Zr]Zr-DOTA chelation can lead to thermal denaturation of the mAb moieties. We developed a method for synthesizing [89Zr]Zr-DOTA-mAb based on a tetrazine (Tz)-conjugated bifunctional DOTA derivative 2,2',2″-(10-(1-(4-(1,2,4,5-tetrazin-3-yl)phenyl)-3,21,26-trioxo-6,9,12,15,18-pentaoxa-29-carboxy-2,22,25-triazanonacosane-29-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (DOTAGA-Tz) and the inverse electron-demand Diels-Alder (IEDDA) click chemistry reaction where trans-cyclooctene-modified mAbs are conjugated to [89Zr]Zr-DOTAGA without being exposed to heat. The stability of IEDDA-derived [89Zr]Zr-DOTAGA-trastuzumab was confirmed by in vitro, ex vivo, and in vivo testing and comparative analysis against the conventional deferoxamine (DFO) counterpart [89Zr]Zr-DFO-trastuzumab. The in vivo immunoPET imaging using [89Zr]Zr-DOTAGA-trastuzumab clearly visualized human epidermal growth factor receptor 2-positive malignancies in murine xenograft models. Greater tumor contrast was observed from [89Zr]Zr-DOTAGA-trastuzumab at a 72-h delayed scan compared with [89Zr]Zr-DFO-trastuzumab. These findings suggest that our IEDDA ligation approach can be an effective means of synthesizing [89Zr]Zr-DOTA-mAb and can enhance the theranostic potential of 89Zr-immunoPET in DOTA-mediated radioimmunotherapy.