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新型放射性治疗 EphA2 靶向双环肽的研发及临床前特征。

Development and preclinical characterization of a novel radiotheranostic EphA2-targeting bicyclic peptide.

机构信息

Department of Nuclear Medicine, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany.

Division of Radiopharmaceutical Development, German Cancer Consortium (DKTK), partner site Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany and German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.

出版信息

Theranostics. 2024 Aug 6;14(12):4701-4712. doi: 10.7150/thno.96641. eCollection 2024.

Abstract

Erythropoietin-producing hepatocellular receptor A2 (EphA2), is a receptor tyrosine kinase involved in cell-cell interactions. It is known to be overexpressed in various tumors and is associated with poor prognosis. EphA2 has been proposed as a target for theranostic applications. Low molecular weight peptide-based scaffolds with low nanomolar affinities have been shown to be ideal in such applications. Bicyclic peptides have emerged as an alternative to traditional peptides for this purpose, offering affinities comparable to antibodies due to their constrained nature, along with high tissue penetration, and improved stability compared to linear counterparts. This study presents the development and comprehensive and preclinical evaluation of BCY18469, a novel EphA2-targeting bicyclic peptide-based radiotheranostic agent. The EphA2-targeting Bicycle peptide BCY18469 was identified through phage-display and chemically optimized. BCY18469 was radiolabeled with Ga, Lu and In. The physicochemical properties, binding affinity and internalization as well as specificity of the peptide were evaluated . PET/MR and SPECT/CT imaging studies were performed using [Ga]Ga-BCY18469 and [In]In-BCY18469, respectively, along with biodistribution of [Lu]Lu-BCY18469 up to 24 h post injection in HT1080- and PC-3-tumor bearing BALB/c nu/nu EphA2-overexpressing xenograft mouse models. The EphA2-targeting bicyclic peptide BCY18469 showed high binding affinity toward human and mouse EphA2 (1.9 and 3.8 nM, respectively). BCY18469 specifically bound and internalized into EphA2-expressing HT1080 cells. Imaging studies showed high tumor enrichment at early time-points (SUV of 1.7 g/mL at 1 h p.i. and 1.2 g/mL at 2 h p.i. in PET/MRI, HT1080 xenograft) with tumor contrast as early as 5 min p.i. and kidney-mediated clearance. Biodistribution studies revealed high early tumor uptake (19.5 ± 3.5 %ID/g at 1 h p.i., HT1080 xenograft) with SPECT/CT imaging further confirming these findings (5.7 ± 1.5 %ID/g at 1 h p.i., PC-3 xenograft). BCY18469 demonstrated high affinity, specific targeting of EphA2, a favorable biodistribution profile, and clearance through renal pathways. These findings underscore the potentially important role of bicyclic peptides in advancing radiotheranostic approaches and encourage additional translational research.

摘要

Erythropoietin-producing hepatocellular receptor A2 (EphA2) 是一种参与细胞间相互作用的受体酪氨酸激酶。它在各种肿瘤中过度表达,与预后不良相关。EphA2 已被提议作为治疗应用的靶点。具有低纳摩尔亲和力的低分子量肽基支架在这种应用中被证明是理想的。双环肽已成为传统肽的替代品,由于其受限的性质,与抗体具有可比的亲和力,同时具有高组织穿透性和与线性对应物相比提高的稳定性。本研究介绍了通过噬菌体展示和化学优化鉴定的新型 EphA2 靶向双环肽放射性治疗药物 BCY18469 的开发和全面的临床前评估。通过噬菌体展示和化学优化鉴定了 EphA2 靶向双环肽 BCY18469。BCY18469 用 Ga、Lu 和 In 进行放射性标记。评估了肽的理化性质、结合亲和力和内化以及特异性。使用 [Ga]Ga-BCY18469 和 [In]In-BCY18469 进行 PET/MR 和 SPECT/CT 成像研究,以及 [Lu]Lu-BCY18469 的生物分布,直至 EphA2 过表达的 HT1080-和 PC-3-肿瘤荷瘤 BALB/c nu/nu 异种移植小鼠模型注射后 24 小时。EphA2 靶向双环肽 BCY18469 对人 EphA2 和小鼠 EphA2 具有高结合亲和力(分别为 1.9 和 3.8 nM)。BCY18469 特异性结合并内化到 EphA2 表达的 HT1080 细胞中。成像研究表明,在早期时间点肿瘤富集程度高(PET/MRI 中 1 小时时的 SUV 为 1.7 g/mL,2 小时时为 1.2 g/mL,HT1080 异种移植),最早在 5 分钟时即可获得肿瘤对比度,随后通过肾脏清除。生物分布研究显示,早期肿瘤摄取率高(1 小时时为 19.5 ± 3.5 %ID/g,HT1080 异种移植),SPECT/CT 成像进一步证实了这一发现(1 小时时为 5.7 ± 1.5 %ID/g,PC-3 异种移植)。BCY18469 表现出高亲和力、特异性靶向 EphA2、良好的生物分布特征以及通过肾脏途径清除。这些发现强调了双环肽在推进放射性治疗方法方面的潜在重要作用,并鼓励进一步的转化研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1786/11373624/764bb1ea3994/thnov14p4701g001.jpg

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