Basic Study for the Design and the Preparation of the Artificial Protein Modules and its Pharmaceutical Applications

人工蛋白模块的设计、制备及其药物应用的基础研究

• 批准号: 17590134
• 负责人: TOMIOKA Yoshihisa
• 金额: $ 2.3万
• 依托单位: Josai International University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590134/
• 关键词: biotechnology; protein modules; pharmaceutical scineces; supernatural product; 生理活性

In order to investigate and/or determine the structure of glycyrrhetic acid (GA) to contribute the binding affinity and specificity against the antibodies, the mouse anti-GA monoclonal antibody (AGA-1) was carefully investigated the binding to the GA-derived compounds, which were 17 compounds tested, with the inhibition ELISA. AGA-1 binding was inhibited by the 16 compounds investigated, except bile acid. We found that the A-ring, the C-3 hydroxyl group, and the C-30 carbonic group for the E-ring, oleanane structure and its C-11 carbonic group were important for the recognition of the AGA-1.We prepared the recombinant Fab fragment antibodies which were substituted the each amino acid in the CDR-H3 with alanine, and investigated the binding activities against the GA-BSA with ELISA. It suggested that the amino acid-side chains for the root region of the CDR-H3 was contributed to the specific binding.In order to investigate the reactivity of the AGA-1 and each rFab, GA was fixed on the sensor chip CM5 for BIACORE and tested. The kinetic parameters were calculated. The parameters kon (E4 /M/s), koff (E4 /M/s), and KD (E-9 M) for AGA-1 were 37.4 31.6, and 3.2, respectively. However, the parameters for the rFab were Y99A (0.9, 0.2, 20.8), Y100bA (36.4, 2.3, 6.2), D101A (16.5, 3.7, 25.1), W95A (5.2, 5.1, 98.4), respectively.

为了研究和/或确定甘草次酸（GA）的结构以有助于对抗体的结合亲和力和特异性，用抑制ELISA仔细研究小鼠抗GA单克隆抗体（阿加-1）与GA衍生化合物（测试的17种化合物）的结合。除胆汁酸外，所研究的16种化合物均抑制阿加-1结合。结果表明，AGA-1的A环、C-3位羟基、C-30位碳原子、齐墩果烷结构及其C-11位碳原子对阿加-1的识别具有重要作用。将AGA-1的CDR-H3中的每个氨基酸替换为丙氨酸，制备了重组Fab片段抗体，并通过ELISA检测了抗体与GA-BSA的结合活性。为了研究阿加-1与每种rFab的反应性，将GA固定在用于BIACORE的传感器芯片CM 5上并进行测试。计算了动力学参数。阿加-1的参数kon（E4 /M/s）、koff（E4 /M/s）和KD（E-9 M）分别为37.4、31.6和3.2。然而，rFab的参数分别为Y 99 A（0.9，0.2，20.8），Y100 bA（36.4，2.3，6.2），D101 A（16.5，3.7，25.1），W 95 A（5.2，5.1，98.4）。

项目成果

Mechanisms of mucin production by rhinovirus infection in cultured human airway epithelial cells

• DOI: 10.1016/j.resp.2005.11.006
• 发表时间: 2006-12-01
• 期刊: RESPIRATORY PHYSIOLOGY & NEUROBIOLOGY
• 影响因子: 2.3
• 作者: Inoue, Daisuke;Yamaya, Mutsuo;Sasaki, Hidetada
• 通讯作者: Sasaki, Hidetada

Preparation of neocarzinostatin apoprotein mutants and its randomized library on the chromophore-binding cavity.

新制癌菌素脱辅基蛋白突变体及其发色团结合腔随机文库的制备。

• 发表时间: 2006
• 期刊: Biol. Pharm. Bull. 29
• 作者: Kragh-Hansen U;Watanabe H;Nakajou K;et al.;山内 あい子;Yoshihisa Tomioka
• 通讯作者: Yoshihisa Tomioka

Preparation of neocarzinostatin apoprotein mutants and the randomized library on the chromophore-binding cavity

新制癌菌素脱辅基蛋白突变体的制备及发色团结合腔随机文库的制备

• 发表时间: 2006
• 期刊: Biological and Pharmaceutical Bulletin 29
• 作者: Kaoru Murata;Nariyasu Mano;Naoki Asakawa;Yasushi Ishihama;Yoshihisa Tomioka
• 通讯作者: Yoshihisa Tomioka

Genetic variation in ABCB1 influences paclitaxel pharmacokinetics in Japanese patients with ovarian cancer

• DOI: 10.1136/ijgc-00009577-200605000-00004
• 发表时间: 2005-12
• 期刊: International Journal of Gynecologic Cancer
• 作者: Hiroaki Yamaguchi;T. Hishinuma;Naomi Endo;Hiroki Tsukamoto;Yukinaga Kishikawa;Mitsumoto Sato;Yuriko Murai;Masahiro Hiratsuka;K. Ito;C. Okamura;Nobuo Yaegashi;N. Suzuki;Yoshihisa Tomioka;Junichi Goto

Preparation of neocarzinostatin apoprotein mutants and its randomized library on the chromophore-binding cavity

新制癌菌素脱辅基蛋白突变体及其发色团结合腔随机文库的制备

• 发表时间: 2006
• 期刊: Biol. Pharm. Bull. 29
• 作者: Iwai;T.;Ito;S.;Tanimitsu;K.;Udagawa;S.;Oka;J.-I.;Yoshihisa Tomioka
• 通讯作者: Yoshihisa Tomioka