7-Fluorosialyl Glycosides Are Hydrolysis Resistant but Readily Assembled by Sialyltransferases Providing Easy Access to More Metabolically Stable Glycoproteins,ACS Central Science

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7-Fluorosialyl Glycosides Are Hydrolysis Resistant but Readily Assembled by Sialyltransferases Providing Easy Access to More Metabolically Stable Glycoproteins
ACS Central Science ( IF 12.7 ) Pub Date : 2021-01-18 , DOI: 10.1021/acscentsci.0c01589
Andreas Geissner ₁ , Lars Baumann ₁ , Thomas J. Morley ₁ , Andrew K. O. Wong ₁ , Lyann Sim ₁ , Jamie R. Rich ₁ , Pauline P. L. So ₂ , Edie M. Dullaghan ₂ , Etienne Lessard ₃ , Umar Iqbal ₃ , Maria Moreno ₃ , Warren W. Wakarchuk ₄ , Stephen G. Withers ₁

Affiliation

The maintenance of therapeutic glycoproteins within the circulatory system is associated, in large part, with the integrity of sialic acids as terminal sugars on the glycans. Glycoprotein desialylation, either by spontaneous cleavage or through host sialidases, leads to protein clearance, mainly through the liver. Thus, the installation of minimally modified sialic acids that are hydrolysis-resistant yet biologically equivalent should lead to increased circulatory half-lives and improved pharmacokinetic profiles. Here we describe the chemoenzymatic synthesis of CMP–sialic acid sugar donors bearing fluorine atoms at the 7-position, starting from the corresponding 4-deoxy-4-fluoro-N-acetylhexosamine precursors. For the derivative with natural stereochemistry we observe efficient glycosyl transfer by sialyltransferases, along with improved stability of the resultant 7-fluorosialosides toward spontaneous hydrolysis (3- to 5-fold) and toward cleavage by GH33 sialidases (40- to 250-fold). Taking advantage of the rapid transfer of 7-fluorosialic acid by sialyltransferases, we engineered the O-glycan of Interferon α-2b and the N-glycans of the therapeutic glycoprotein α1-antitrypsin. Studies of the uptake of the glyco-engineered α1-antitrypsin by HepG2 liver cells demonstrated the bioequivalence of 7-fluorosialic acid to sialic acid in suppressing interaction with liver cell lectins. In vivo pharmacokinetic studies reveal enhanced half-life of the protein decorated with 7-fluorosialic acid relative to unmodified sialic acid in the murine circulatory system. 7-Fluorosialylation therefore offers considerable promise as a means of prolonging circulatory half-lives of glycoproteins and may pave the way toward biobetters for therapeutic use.

中文翻译：

7-氟代唾液酸糖苷具有抗水解性，但易于通过唾液酸转移酶组装，从而可轻松获得更多代谢稳定的糖蛋白

循环系统中治疗性糖蛋白的维持在很大程度上与唾液酸作为聚糖末端糖的完整性有关。糖蛋白的去唾液酸化作用（通过自然裂解或通过宿主唾液酸酶）可导致蛋白质清除，主要是通过肝脏。因此，安装具有抗水解性但生物学等效性的最低限度修饰的唾液酸应可提高循环半衰期并改善药代动力学。在这里，我们描述了在7位上带有氟原子的CMP-唾液酸糖供体的化学酶法合成，从相应的4-deoxy-4-fluoro- N开始-乙酰己糖胺前体。对于具有自然立体化学的衍生物，我们观察到通过唾液酸转移酶进行的有效糖基转移，以及所得7-氟唾液酸苷对自发水解（3至5倍）和被GH33唾液酸酶裂解（40至250倍）的稳定性提高。利用唾液酸转移酶快速转移7-氟唾液酸的优势，我们设计了干扰素α-2b的O-聚糖和治疗性糖蛋白α1-抗胰蛋白酶的N-聚糖。HepG2肝细胞摄取糖工程化的α1-抗胰蛋白酶的研究表明，在抑制与肝细胞凝集素的相互作用中，7-氟唾液酸与唾液酸具有生物等效性。体内药代动力学研究表明，相对于未修饰的唾液酸，在鼠类循环系统中，用7-氟唾液酸修饰的蛋白的半衰期延长。因此，7-氟唾液酰化作为延长糖蛋白循环半衰期的手段提供了相当可观的前景，并可能为治疗性生物制剂铺平道路。

更新日期：2021-02-24

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