A study on the application of hemoglobin derivatives that possess nitric oxide releasing capacity as artificial red cells

具有一氧化氮释放能力的血红蛋白衍生物作为人工红细胞的应用研究

• 批准号: 11694229
• 负责人: KITABATAKE Akira
• 金额: $ 10.54万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11694229/
• 关键词: artificial red blood cell; hemoglobin derivative; nitric oxide; platelet aggregation; platelet adhesion; polyethylene glycol; s-nitrosohemoglobin; perfluorocarbon; セロトニン

A cellular hemoglobin (Hb) derivatives developed as red blood cell substitutes (BCSs) are known to be potent vasoconstrictors due to their EDRF/NO scavenging action. We therefore developed polyethylene glycol-modified Hb (PEG-Hb), and further modified it to make s-nitrosylated PEG-Hb (SNO-PEG-Hb) as new candidate oxygen carriers. SNO-PEG-Hb possesses desirable properties as an BCS since it can release NO when tissue is hypoxic and absorbs NO when NO is produced to much. Furthermore, using a new machine we synthesized new emulsion to perfluorochemicals (PFCs) and manufactured several PFC compound as BCSs. We found that after injecting those compound to the rat that SNO-PEG-Hb induced least effects on functions and histology of liver and kidney among three Hb-based oxygen carriers (HBOC)s. Using flowcytemetry effects of HBOCs and PFC compounds on platelets were checked and t was suggested that SNO-PEG-Hb and the new PFC compound have least effects. We tried the application ofHBOCs on acute anemia induced by Babesia infection in the dog. When SNO-PEG-Hb was infused to the brain ischemia-reperfusion model of the rats, it ameliorated their cognitive function. We applied the PFC compound in a dog model of cardiopulmonary bypass operation and it revealed that the compound can save heart from anemia-induced hypoxia. On this occasion simultaneous infusion of albumin was necessary. The ability of PFC compound for the preservation material in liver transplantation was checked. The acute toxicological test of PFCs compounds were performed and it was suggested that the new compound was the safest. Thus, SNO-PEG-Hb and the new PFC compound are expected to be good candidates for clinically available BCSs.

作为红细胞替代品(Bcs)的细胞血红蛋白(Hb)衍生物由于其EDRF/NO清除作用而被认为是有效的血管收缩药。因此，我们开发了聚乙二醇改性的Hb(聚乙二醇化Hb)，并对其进行了进一步的修饰，使其成为新的候选氧载体S硝基聚乙二醇化Hb(SNO-PEGHb)。SnO-PEG-HB作为BCS具有理想的性质，因为它可以在组织缺氧时释放NO，并在NO产生过多时吸收NO。此外，我们利用一台新的设备合成了新的乳液来合成全氟化合物(PFC)，并制备了几种PFC化合物作为BCS。我们发现，在三种含氧载体中，SNO-PEG-Hb对大鼠肝肾功能和组织学的影响最小，用流式细胞术检测了HBOC和PFC化合物对大鼠血小板的影响，提示SNO-PEG-Hb和新的PFC化合物对大鼠的影响最小。我们尝试了HBOCs在犬巴贝斯虫感染所致急性贫血中的应用。在大鼠脑缺血再灌流模型中注入SNO-PEG-Hb，可改善其认知功能。我们将PFC化合物应用于犬体外循环手术模型，结果表明，该化合物可以保护心脏免受贫血所致的缺氧。在这种情况下，同时输注白蛋白是必要的。检测PFC复合体作为肝移植保存材料的能力。对全氟碳化物化合物进行了急性毒性试验，结果表明新化合物是最安全的。因此，SNO-PEG-HB和新的PFC化合物有望成为临床上可用的BCS的良好候选者。

项目成果

Nakal K, Sakuma I, Togashi H, Yoshioka M, Satoh H, Kitabatake A: "S-Nitrosylated polyethylene glycol-conjugated hemoglobin derivative as a candidate material for oxygen theraneutics. In : Maeda H. editor. Polymer drugs in the clinical stage : Advantages a

Nakal K、Sakuma I、Togashi H、Yoshioka M、Satoh H、Kitabatake A：“S-亚硝基化聚乙二醇缀合血红蛋白衍生物作为氧治疗的候选材料。见：Maeda H.编辑。临床阶段的聚合物药物：

藤井ひとみ ほか: "赤血球代替物性能評価の方法の開発と臨床試験"人工血液. 7(1). 7-15 (1999)

Hitomi Fujii 等人：“评估红细胞替代品性能的方法的开发和临床试验”Artificial Blood 7(1) (1999)。

Ichiro Sakuma, et al.: "Application of SNO-hemoglobin as an Artificial oxygen carrier"Vessels and Endothelium. 9(Suppl.). 71-82 (1999)

Ichiro Sakuma 等人：“SNO-血红蛋白作为人工氧载体的应用”血管和内皮。

佐久間一郎、仲井邦彦、菅原 武、佐藤 洋、北畠 顕: "s-ニトロソヘモグロビンの基礎と臨床"THE LUNG perspectives. 9(2). 191-194 (2001)

Ichiro Sakuma、Kunihiko Nakai、Takeshi Sugara、Hiroshi Sato、Akira Kitabatake：“S-亚硝基血红蛋白的基础和临床实践”THE LUNG 观点 9(2)。

Fujii H, Fujii S, et al.: "Attenuation of hypothermia-induced platelet activation and platelet adhesion to artificial surfaces in vitro by modification of hemoglobin to carry S-nitric oxide and polyethylene glycol."Thrombosis Research. 100(6). 519-528 (20

Fujii H、Fujii S 等人：“通过修饰血红蛋白携带 S-一氧化氮和聚乙二醇，减弱低温诱导的血小板活化和血小板在体外对人造表面的粘附。”血栓形成研究。