Design and Evaluation of Albumin Mutants for Clinical Application

临床应用白蛋白突变体的设计和评价

• 批准号: 11694298
• 负责人: OTAGIRI Masaki
• 金额: $ 4.67万
• 依托单位: KUMAMOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694298/
• 关键词: Albumin preparation; Site-directed mutagenesis; Molecular design; Artificial blood; Functional evaluation; Blood preparation; Clinical application

International collaboration work for reducing the rate of albumin preparations usage was undertaken to develop a safe and effective recombinant albumins. The results obtained are as follow :1) Two single-mutants (K199A, K525A) were produced. The recombinant albumins were correctly folded, as evidenced by the fact that they showed similar patterns as the native albumin in the far- and near-UV CD spectrum as well as reactivity towards polyclonal anti-serum raised against native albumin.2) Thermal denaturation experiments by using GdnCl and DSC revealed no significant difference in conformational stability for native and mutant albumins.3) The ligand binding properties and esterase-like activity of the albumin mutants were nearly the same as those for the native albumin.4) The plasma levels of the albumin mutants labeled with ^<125>I after intravenous bolus injections to rats showed two-phase profiles but their half-lives were shorten than that for the native albumin.5) Dr.Kragh-Hansen of Aarhus University visited Kumamoto University twice (Oct.23-Nov.7, 1999 ; May 5-Oct.29, 2000) to discuss about pharmacokinetic and biological properties of the albumin mutants. Dr.Curry of Imperial College also visited Kumamoto University (Sep.27-Oct.26, 2000) to discuss about structural characterization of the albumin mutants.

为了降低白蛋白制剂速率的国际协作工作，以开发安全有效的重组白蛋白。获得的结果如下：1）产生了两种单突（K199A，K525A）。重组白蛋白被正确折叠了，这是事实证明的：它们在远处和近紫外CD频谱中表现出与本机白蛋白相似的模式，以及对针对天然白蛋白的多克隆抗苏联的反应性。2）使用GDNCL和DSC中的构型构型和变形符中没有显着差异。白蛋白突变体的类似酯酶的活性几乎与本地白蛋白的活性几乎相同。4）对大鼠静脉注射后标记为 ^<125> i的白蛋白突变体的血浆水平显示出两相的特征，但其半衰期比本地白蛋白的半衰期较短。5） （1999年10月23日； 1999年； 5月5日至29日，2000年）讨论白蛋白突变体的药代动力学和生物学特性。帝国学院的库尔里博士还访问了库曼本大学（9月27日至26日，2000年），讨论白蛋白突变体的结构表征。

项目成果

Sakai T, et al.: "Interaction mechanism between indoxyl sulfate, a typical uremic toxin bound to site II, and ligands bound to site I of human serum albumin."Pharm Res.. (in press). (2001)

Sakai T 等人：“硫酸吲哚酚（一种与位点 II 结合的典型尿毒症毒素）与与人血清白蛋白位点 I 结合的配体之间的相互作用机制。”Pharm Res..（出版中）。

V.T.G.Chuang et al: "Helix of subdomain IIIA of HSA is the region primainly photolabelet by betoproten"Biochimica Biophysica Acta. 1434. 18-30 (1999)

V.T.G.Chuang 等人：“HSA 子结构域 IIIA 的螺旋是主要由 Betoproten 进行光标记的区域”Biochimica Biophysicala Acta。

Y.Tsutsumi: "Decreased bilirubin-binding capacity in uremicserum caused by an accumulation of furan dicarboxylic acid"Nephrom. 84・(3)(in press). (2000)

Y. Tsutsumi：“呋喃二羧酸积累导致尿毒症血清胆红素结合能力降低”Nephrom 84・(3)（出版中）。

T.Sakai et al: "Stereoselective serum protein binding of be to profen in liver diseises"Enantiomer. 4・(5). 477-482 (1999)

T.Sakai 等人：“肝脏疾病中 Be 与洛芬的立体选择性血清蛋白结合”4·(5)477-482 (1999)。

T.Sakai et al: "Interaction between indoxyl sulfate, a typical uremic toxin bound to site II, and ligands bound to site I of HSA"Pharmaceutical Research. 18(4)(in press). (2001)

T.Sakai 等人：“硫酸吲哚酚（一种与位点 II 结合的典型尿毒症毒素）与与 HSA 位点 I 结合的配体之间的相互作用”药物研究。