鋅離子誘導之聚組氨酸融合蛋白及胜肽自組裝於蛋白質藥物釋放之調控

Abstract

近年來，蛋白質微米顆粒於藥物遞送系統的潛在應用倍受矚目，不但能提高蛋白質的穩定性，並可延長體內的釋放時間。然而，大多數研究的製備方式涉及交聯或高鹽等過程，常導致蛋白質失活並且難以用於蛋白質藥物的釋放系統上。本研究利用已發展成熟的六聚組氨酸標記融合蛋白技術以及金屬可調控胜肽，來強化蛋白質與金屬離子之間的作用能力並藉此調整所包覆之蛋白質藥物釋放速率。於鋅離子(1 mM)添加後，六聚組氨酸標記融合蛋白可自組裝形成直徑約兩微米的蛋白質微米顆粒，並可於生理食鹽水中持續釋放八小時。實驗結果指出，藉由調整胜肽濃度與蛋白質兩端六聚組氨酸標記，能合成出可控制釋放速率之蛋白質藥物載體，並且，在模型藥物的釋放實驗中，並無觀察到蛋白質結構的破壞。本研究開發一簡易、條件溫和之通用蛋白質微米顆粒合成技術並成功應用於可調控的蛋白質藥物釋放系統。

In recent years, protein microparticles have received much attention in the drug delivery systems due to the improved protein stability and prolonged release in vivo. However, most of current preparation processes involved harsh conditions, introducing chemical crosslinkers or high salt, which often led to protein inactivation and failed to apply in drug delivery systems. In this study, we employed the well-established hexahistidine (His)-tag recombinant protein technology as well as a metal-triggerable peptide to enhance the binding strength between protein and metal ion and to fine-tune the protein drug release. The His-tagged proteins could self-assemble to form microparticles (~ 2 μm) upon zinc chloride (1 mM) treatment and an eight-hour sustained protein drug release has been achieved in physiological saline. The experimental results also indicated that by adjusting the peptide concentration and the N- and C-terminal hexahistidine-tags, the protein release could be controlled. Moreover, no protein denaturation has been observed. We have developed a universal strategy to enable facile protein microparticles fabrication under mild conditions, and their potential in release-tunable protein drug delivery systems has been successfully demonstrated.