Research Division, Chugai Pharmaceutical Co., Ltd., Gotemba, Shizuoka 412-8513, Japan.
Research Division, Chugai Pharmaceutical Co., Ltd., Gotemba, Shizuoka 412-8513, Japan.
Toxicol In Vitro. 2020 Aug;66:104841. doi: 10.1016/j.tiv.2020.104841. Epub 2020 Apr 1.
An emerging structure for anti-tumor antibody drugs utilizes a bispecific antibody (BiAb) that recognizes a tumor surface antigen and CD3 on T cells. An impurity that commonly contaminates these BiAb products is an anti-CD3 monoclonal antibody (mAb). The most plausible cause of toxic activity by an anti-CD3 mAb is the induction of cytokines via T cell activation. In this in vitro study, we compared cytokine induction and T cell activation after treatment with an anti-glypican-3/CD3 BiAb (ERY974), anti-CD3 mAb impurity (aCD3), or ERY974 spiked with 5% aCD3. We found that contamination with up to 5% aCD3 did not affect cytokine release by ERY974. Cytokine levels induced by ERY974 in the presence of target cells were significantly higher than those induced by aCD3, but were very similar to those by the spiked treatment. The results supported the specification of a 5% limit for aCD3. OKT-3 had much higher activity to induce cytokines from peripheral blood mononuclear cells in an in vitro assay than aCD3. This suggests that specification limit should be decided for each type of anti-CD3 impurity that affects T cell-activating BiAb drug products. In vitro cytokine assays can provide useful information for determining these specification limits.
一种新兴的抗肿瘤抗体药物结构利用双特异性抗体（BiAb），该抗体识别肿瘤表面抗原和 T 细胞上的 CD3。这些 BiAb 产品中常见的杂质是抗 CD3 单克隆抗体（mAb）。抗 CD3 mAb 产生毒性作用的最可能原因是通过 T 细胞激活诱导细胞因子。在这项体外研究中，我们比较了在用抗聚糖蛋白 3/CD3 BiAb（ERY974）、抗 CD3 mAb 杂质（aCD3）或添加 5% aCD3 的 ERY974 处理后细胞因子诱导和 T 细胞激活。我们发现，含有高达 5%的 aCD3 不会影响 ERY974 释放细胞因子。在存在靶细胞的情况下，ERY974 诱导的细胞因子水平明显高于 aCD3 诱导的细胞因子水平，但与添加处理诱导的细胞因子水平非常相似。结果支持了 aCD3 杂质含量为 5%的规定。在体外测定中，OKT-3 诱导外周血单个核细胞释放细胞因子的活性比 aCD3 高得多。这表明，应针对每种影响 T 细胞激活 BiAb 药物产品的抗 CD3 杂质来确定规定限值。体外细胞因子测定可为确定这些规定限值提供有用信息。
