Development of Long-Acting Human Adrenomedullin Fc-Fusion Proteins.
Development of Long-Acting Human Adrenomedullin Fc-Fusion Proteins.
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DOI：
10.3390/biology11071074
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发表时间：
2022-07-19
期刊：
影响因子：
4.2
通讯作者：
Kitamura K
中科院分区：
文献类型：
--
作者：
Nagata S;Yamasaki M;Kuroishi N;Kitamura K
肾上腺髓质素 (AM) 是一种降血压肽激素，具有抗炎作用，并参与伤口愈合和胚胎发生。然而，由于天然AM在血液中的半衰期很短，因此治疗需要连续给药。为了解决这个问题，我们使用哺乳动物细胞开发了四种含有全长 hAM 或 hAM 残基 6-52 的人 IgG1 和 IgG4 Fc 融合蛋白。产生的 Fc-AM 被酰胺化并处于活性形式。所有 Fc-AM 均刺激稳定表达 AM1 受体的 HEK-293 细胞中 cAMP 的产生。 IgG1-AM (6-52)和IgG4-AM (6-52)的活性高于IgG1-AM和IgG4-AM。单次皮下给药后 14 天，在血液中观察到足够浓度的 IgG1-AM (6-52) 和 IgG4-AM (6-52)。此外，给予IgG1-AM(6-52)或IgG4-AM(6-52)后，观察到组织转移至肾脏和小肠。 IgG4-AM (6-52) 治疗可抑制自发性高血压大鼠的血压升高。由哺乳动物细胞产生的 Fc-AM 易于制备，可能是一种有效的新型治疗剂。 (1)背景：人肾上腺髓质素(hAM)是一种降压肽激素，具有强大的抗炎作用。 AM还在各种疾病动物实验模型中具有治疗作用。然而，由于天然AM在血液中的半衰期很短，因此治疗需要连续给药。为了解决这个问题，我们开发了四种含有全长 hAM 或 hAM 残基 6-52 的人 IgG1 和 IgG4 Fc 融合蛋白。 (2)方法：利用哺乳动物细胞制备重组Fc-AM衍生物，并测试Fc-AM的药代动力学和生物活性。 （3）结果：我们开发了四种Fc融合AM（Fc-AM），它们是哺乳动物细胞中的长效AM衍生物。 Fc-AM 在血液中的半衰期较长，并保留了与 AM1 受体结合的能力。 Fc-AM (6-52) 比 Fc-AM 诱导更高的受体 cAMP 水平。给大鼠施用IgG1-AM（6-52）或IgG4-AM（6-52）后，观察到组织转移至肾脏和小肠。此外，用 IgG4-AM (6-52) 治疗可抑制自发性高血压大鼠的血压升高。 (4)结论：哺乳动物细胞产生的Fc-AM易于制备，可能是一种有效的新型治疗剂。
Adrenomedullin (AM) is a hypotensive peptide hormone that exerts anti-inflammatory effects and is involved in wound healing and embryogenesis. However, treatment requires continuous administration as the half-life of native AM is short in blood. To resolve this, we developed four human IgG1 and IgG4 Fc-fusion proteins containing full-length hAM or hAM residues 6-52 using mammalian cells. The Fc-AM produced were amidated and in the active form. All Fc-AMs stimulated cAMP production in HEK-293 cells stably expressing the AM1 receptor. The activities of IgG1-AM (6-52) and IgG4-AM (6-52) were higher than those of IgG1-AM and IgG4-AM. Sufficient concentrations of IgG1-AM (6-52) and IgG4-AM (6-52) were observed in blood 14 days after a single subcutaneous administration. Furthermore, after IgG1-AM (6-52) or IgG4-AM (6-52) administration, tissue transfer to the kidney and small intestine was observed. Treatment with IgG4-AM (6-52) inhibited blood pressure increase in spontaneously hypertensive rats. Fc-AM produced from mammalian cells can be easily prepared and might be an effective novel therapeutic agent. (1) Background: Human adrenomedullin (hAM) is a hypotensive peptide hormone that exerts powerful anti-inflammatory effects. AM also had therapeutic effects in various animal experimental models of disease. However, treatment required continuous administration as the half-life of native AM is short in blood. To resolve this, we developed four human IgG1 and IgG4 Fc-fusion proteins containing full-length hAM or hAM residues 6-52. (2) Methods: We used mammalian cells to produce recombinant Fc-AM derivatives and tested the pharmacokinetics and biological activity of Fc-AM. (3) Results: We developed four Fc-fusion AMs (Fc-AM), which are long-acting AM derivatives in mammalian cells. Fc-AM had a prolonged half-life in blood and retained its ability to bind to the AM1 receptor. Fc-AM (6-52) induced higher cAMP levels for the receptor than Fc-AM. After the administration of IgG1-AM (6-52) or IgG4-AM (6-52) to rats, tissue transfer to the kidney and small intestine was observed. In addition, treatment with IgG4-AM (6-52) inhibited blood pressure increase in spontaneously hypertensive rats. (4) Conclusions: Fc-AM produced from mammalian cells can be easily prepared and might be an effective novel therapeutic agent.
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影响因子：
4.8
作者：
Kuwasako, K;Shimekake, Y;Sakata, T
通讯作者：
Sakata, T
影响因子：
3.4
作者：
Serrano-Ponz M;Rodrigo-Gasqué C;Siles E;Martínez-Lara E;Ochoa-Callejero L;Martínez A
通讯作者：
Martínez A
影响因子：
3
作者：
Hay, DL;Smith, DM
通讯作者：
Smith, DM
影响因子：
4.8
作者：
Kita, Toshihiro;Kaji, Yoshikazu;Kitamura, Kazuo
通讯作者：
Kitamura, Kazuo
影响因子：
38.9
作者：
Laterre PF;Pickkers P;Marx G;Wittebole X;Meziani F;Dugernier T;Huberlant V;Schuerholz T;François B;Lascarrou JB;Beishuizen A;Oueslati H;Contou D;Hoiting O;Lacherade JC;Chousterman B;Pottecher J;Bauer M;Godet T;Karakas M;Helms J;Bergmann A;Zimmermann J;Richter K;Hartmann O;Pars M;Mebazaa A;AdrenOSS-2 study participants
通讯作者：
AdrenOSS-2 study participants