Semisynthesis of natural glycoforms of human erythropoietin

人促红细胞生成素天然糖型的半合成

• 批准号: 223435149
• 负责人: Professor Dr. Carlo Unverzagt
• 金额: --
• 依托单位: Arbeitsgruppe Bioorganische Chemie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/223435149?language=en
• 关键词: Semisynthesis; natural; glycoforms; human; erythropoietin

Due to its medical importance erythropoietin (EPO) is one of the best-studied recombinant therapeutic glycoproteins. In seminal studies it was revealed that the function of erythropoietin is highly dependent on the presence and the structure of its carbohydrate moieties. Despite many efforts the isolation of homogeneous EPO glycoforms is still not feasible and biological studies can only use mixtures of glycoforms. Thus, synthetic alternatives are desirable aiming at the generation of pure EPO glycoforms needed for detailed structure-activity-relationships. We have established a semisynthetic route to bioactive EPO with an Asn 24 N-glycan using a newly developed convergent synthesis for glycopeptides with all natural linkages. The glycan of Asn 24 can thus be varied readily in order to correlate systematically varied carbohydrate structures to the bioactivity of the protein, its intrinsic stability and the propensity to crystallize. A synthetic access to glycoforms of fully N- and O-glycosylated EPO will also be investigated. The systematic library of variants of natural glycoforms of EPO should provide general insights into the roles of carbohydrates in glycoproteins and may guide the future engineering of glycoforms.

促红细胞生成素(EPO)具有重要的医学价值，是目前研究最多的重组治疗性糖蛋白之一。在开创性的研究中发现，促红细胞生成素的功能高度依赖于其碳水化合物部分的存在和结构。尽管做了很多努力，均一的EPO糖体的分离仍然是不可行的，生物学研究只能使用糖体的混合物。因此，合成替代品是可取的，目的是产生详细的结构-活性-关系所需的纯EPO糖形。我们利用一种新开发的具有全天然键的糖肽的收敛合成方法，建立了一条以天冬氨酸24N-聚糖为原料的半合成EPO的路线。因此，ASN 24的多糖可以很容易地改变，以便系统地将不同的碳水化合物结构与蛋白质的生物活性、其内在稳定性和结晶倾向相关联。还将研究合成完全N-和O-糖基化的EPO糖体的途径。EPO天然糖型变异体的系统文库应该提供对糖蛋白中碳水化合物作用的一般见解，并可能指导未来的糖型工程。