Department of Radiology , Memorial Sloan Kettering Cancer Center (MSK) , New York , NY 10065 , United States.
Molecular Pharmacology Program , MSK , New York , NY 10065 , United States.
Mol Pharm. 2018 Jun 4;15(6):2133-2141. doi: 10.1021/acs.molpharmaceut.7b01133. Epub 2018 Apr 30.
Antibodies labeled with positron-emitting isotopes have been used for tumor detection, predicting which patients may respond to tumor antigen-directed therapy, and assessing pharmacodynamic effects of drug interventions. Prolactin receptor (PRLR) is overexpressed in breast and prostate cancers and is a new target for cancer therapy. We evaluated REGN2878, an anti-PRLR monoclonal antibody, as an immunoPET reagent. REGN2878 was labeled with Zr-89 after conjugation with desferrioxamine B or labeled with I-131/I-124. In vitro determination of the half-maximal inhibitory concentration (IC50) of parental REGN2878, DFO-REGN2878, and iodinated REGN2878 was performed by examining the effect of the increasing amounts of these on uptake of trace-labeled I-131 REGN2878. REGN1932, a non-PRLR binding antibody, was used as a control. Imaging and biodistribution studies were performed in mice bearing tumor xenografts with various expression levels of PRLR, including MCF-7, transfected MCF-7/PRLR, PC3, and transfected PC3/PRLR and T4D7v11 cell lines. The specificity of uptake in tumors was evaluated by comparing Zr-89 REGN2878 and REGN1932, and in vivo competition compared Zr-89 REGN2878 uptake in tumor xenografts with and without prior injection of 2 mg of nonradioactive REGN2878. The competition binding assay of DFO-REGN2878 at ratios of 3.53-5.77 DFO per antibody showed IC50 values of 0.4917 and 0.7136 nM, respectively, compared to 0.3455 nM for parental REGN2878 and 0.3343 nM for I-124 REGN2878. Imaging and biodistribution studies showed excellent targeting of Zr-89 REGN2878 in PRLR-positive xenografts at delayed times of 189 h (presented as mean ± 1 SD, percent injected activity per mL (%IA/mL) 74.6 ± 33.8%IA/mL). In contrast, MCF-7/PRLR tumor xenografts showed a low uptake (7.0 ± 2.3%IA/mL) of control Zr-89 REGN1932 and a very low uptake and rapid clearance of I-124 REGN2878 (1.4 ± 0.6%IA/mL). Zr-89 REGN2878 has excellent antigen-specific targeting in various PRLR tumor xenograft models. We estimated, using image-based kinetic modeling, that PRLR antigen has a very rapid in vivo turnover half-life of ∼14 min from the cell membrane. Despite relatively modest estimated tumor PRLR expression numbers, PRLR-expressing cells have shown final retention of the Zr-89 REGN2878 antibody, with an uptake that appeared to be related to PRLR expression. This reagent has the potential to be used in clinical trials targeting PRLR.
用正电子发射同位素标记的抗体已被用于肿瘤检测,以预测哪些患者可能对肿瘤抗原导向治疗有反应,并评估药物干预的药效学效应。催乳素受体(PRLR)在乳腺癌和前列腺癌中过度表达,是癌症治疗的新靶点。我们评估了 REGN2878,一种抗 PRLR 单克隆抗体,作为免疫 PET 试剂。REGN2878 在与去铁胺 B 缀合后用 Zr-89 标记,或用 I-131/I-124 标记。通过检查这些物质的浓度逐渐增加对放射性标记的 I-131 REGN2878 摄取的影响,测定亲本 REGN2878、DFO-REGN2878 和碘代 REGN2878 的半最大抑制浓度(IC50)。REGN1932,一种非 PRLR 结合抗体,用作对照。在具有不同 PRLR 表达水平的异种移植肿瘤小鼠中进行了成像和生物分布研究,包括 MCF-7、转染的 MCF-7/PRLR、PC3 和转染的 PC3/PRLR 和 T4D7v11 细胞系。通过比较 Zr-89 REGN2878 和 REGN1932,评估了肿瘤摄取的特异性,并通过比较肿瘤异种移植中 Zr-89 REGN2878 的摄取,评估了体内竞争在没有预先注射 2mg 非放射性 REGN2878 的情况下。在 3.53-5.77 DFO 与抗体的比率下,DFO-REGN2878 的竞争结合测定显示 IC50 值分别为 0.4917 和 0.7136 nM,而亲本 REGN2878 为 0.3455 nM,I-124 REGN2878 为 0.3343 nM。成像和生物分布研究表明,在延迟 189 小时时,Zr-89 REGN2878 在 PRLR 阳性异种移植中具有出色的靶向性(表示为平均值±1 SD,每毫升注入的放射性活度(%IA/mL)74.6±33.8%IA/mL)。相比之下,MCF-7/PRLR 肿瘤异种移植对对照 Zr-89 REGN1932 的摄取率较低(7.0±2.3%IA/mL),而 I-124 REGN2878 的摄取率非常低且清除速度很快(1.4±0.6%IA/mL)。Zr-89 REGN2878 在各种 PRLR 肿瘤异种移植模型中具有出色的抗原特异性靶向性。我们使用基于图像的动力学建模估计,PRLR 抗原在体内从细胞膜的半衰期非常快,约为 14 分钟。尽管肿瘤 PRLR 表达的估计数量相对较小,但表达 PRLR 的细胞仍保留了 Zr-89 REGN2878 抗体,其摄取似乎与 PRLR 表达有关。该试剂有可能用于针对 PRLR 的临床试验。