The effects of arginine glutamate, a promising excipient for protein formulation, on cell viability: Comparisons with NaCl.
The effects of arginine glutamate, a promising excipient for protein formulation, on cell viability: Comparisons with NaCl.
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DOI：
10.1016/j.tiv.2016.02.002
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发表时间：
2016-06
期刊：
影响因子：
--
通讯作者：
Dearman RJ
中科院分区：
文献类型：
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作者：
Kheddo P;Golovanov AP;Mellody KT;Uddin S;van der Walle CF;Dearman RJ
在单克隆抗体制剂的背景下，参考NaCl，使用体外细胞培养系统检查了l-精氨酸和l-谷氨酸（Arg·Glu）的等摩尔混合物对细胞活力和细胞应激的影响。选择与皮下给药相关的细胞：人单核细胞系THP-1，作为单细胞悬液生长，和贴壁人原代成纤维细胞。对于THP-1细胞，研究了由相对高的盐浓度引起的细胞死亡的机制，并评估了对细胞活化/应激的影响，作为膜标志物和细胞因子（白细胞介素-8）表达变化的函数。这些研究表明，与相同渗透压的NaCl相比，Arg·Glu对THP-1活力没有任何进一步的有害影响，并且两种盐在较高浓度下均通过凋亡引起细胞死亡;对THP-1细胞应激/活化的测量没有显著影响。对于粘附的成纤维细胞，这两种盐在~ 400 mOsm/kg时引起显著毒性，尽管Arg·Glu引起的随后活力下降比NaCl更急剧。这些数据表明，Arg·Glu的毒性与NaCl相当，并且毒性机制是这样的，即细胞死亡不太可能在体内皮下注射后引发炎症。以NaCl为对照，观察Arg·Glu对细胞活力的影响。Arg·Glu的毒性与NaCl相当。细胞死亡的机制是通过细胞凋亡，因此不是促炎性的。Arg·Glu作为亚切辅料预期不会导致非典型炎症。
The effects of an equimolar mixture of l-arginine and l-glutamate (Arg·Glu) on cell viability and cellular stress using in vitro cell culture systems are examined with reference to NaCl, in the context of monoclonal antibody formulation. Cells relevant to subcutaneous administration were selected: the human monocyte cell line THP-1, grown as a single cell suspension, and adherent human primary fibroblasts. For THP-1 cells, the mechanism of cell death caused by relatively high salt concentrations was investigated and effects on cell activation/stress assessed as a function of changes in membrane marker and cytokine (interleukin-8) expression. These studies demonstrated that Arg·Glu does not have any further detrimental effects on THP-1 viability in comparison to NaCl at equivalent osmolalities, and that both salts at higher concentrations cause cell death by apoptosis; there was no significant effect on measures of THP-1 cellular stress/activation. For adherent fibroblasts, both salts caused significant toxicity at ~ 400 mOsm/kg, although Arg·Glu caused a more precipitous subsequent decline in viability than did NaCl. These data indicate that Arg·Glu is of equivalent toxicity to NaCl and that the mechanism of toxicity is such that cell death is unlikely to trigger inflammation upon subcutaneous injection in vivo. The effect of Arg·Glu on cell viability was examined with reference to NaCl. Arg·Glu is of equivalent toxicity to NaCl. The mechanism of cell death is via apoptosis and is thus not pro-inflammatory. Arg·Glu as a subcut excipient is not expected to result in atypical inflammation.