Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan.
Nitric Oxide. 2010 Sep 15;23(2):121-7. doi: 10.1016/j.niox.2010.05.002. Epub 2010 May 6.
S-Nitrosated human serum albumin (SNO-HSA) is a large molecular weight nitric oxide carrier in human plasma, and because of its many beneficial effects in different tests, it is currently under investigation as a cytoprotective agent. However, making SNO-HSA preparations is a complicated and time-consuming process. We found that binding of caprylic acid (CA) and N-acetyl-l-tryptophan (N-AcTrp) to defatted mercaptalbumin increased S-nitrosation by S-nitrosoglutathione (GS-NO) by making Cys-34 of HSA more accessible and by protecting it against oxidation, respectively. Fortunately, HSA solutions for clinical use contain high concentrations of CA and N-AcTrp as stabilizers. By making use of that fact it was possible to work-out a fast and simple procedure for producing SNO-HSA: incubation of a commercial HSA formulation with GS-NO for only 1 min results in S-nitrosation of HSA. The biological usefulness of such a preparation was tested in a rat ischemia-reperfusion liver injury model. Although our procedure for making SNO-HSA is fast and straightforward, the cytoprotective effect of the preparation was similar to, or better than, that of a preparation made in a more traditional way. The clinical development of SNO-HSA as a strong cytoprotective agent is under way using this method in collaboration with clinicians and industrial developers.
人血清白蛋白的 S-亚硝基化产物（SNO-HSA）是人类血浆中一种大分子量的一氧化氮载体，由于其在不同试验中具有许多有益的作用，目前正在作为细胞保护剂进行研究。然而，制备 SNO-HSA 制剂是一个复杂且耗时的过程。我们发现，辛酸（CA）和 N-乙酰色氨酸（N-AcTrp）与脱脂巯基白蛋白结合，分别通过使 HSA 中的 Cys-34 更易接近和防止其氧化，增加了 S-亚硝基谷胱甘肽（GS-NO）的 S-亚硝基化。幸运的是，用于临床的 HSA 溶液含有高浓度的 CA 和 N-AcTrp 作为稳定剂。利用这一事实，我们得以制定出一种快速而简单的制备 SNO-HSA 的方法：只需将商业 HSA 制剂与 GS-NO 孵育 1 分钟，即可使 HSA 发生 S-亚硝基化。在大鼠缺血再灌注肝损伤模型中测试了这种制剂的生物学用途。尽管我们制备 SNO-HSA 的方法快速而直接，但该制剂的细胞保护作用与以更传统方式制备的制剂相似，甚至更好。该方法与临床医生和工业开发者合作，正在将 SNO-HSA 作为一种强大的细胞保护剂进行临床开发。
