C-Mannosyl Lysine for Solid Phase Assembly of Mannosylated Peptide Conjugate Cancer Vaccines
C-Mannosyl Lysine for Solid Phase Assembly of Mannosylated Peptide Conjugate Cancer Vaccines
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DOI：
10.1021/acschembio.9b00987
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发表时间：
2020-03-20
影响因子：
4
通讯作者：
Codee, Jeroen D. C.
中科院分区：
文献类型：
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作者：
Hogervorst, Tim P.;Li, R. J. Eveline;Codee, Jeroen D. C.
树突状细胞（DC）装备有大量模式识别受体（PRR）以识别病原体并启动病原体定制的T细胞应答。在这些反应中，DC的成熟是关键，以及有助于完成T细胞反应的细胞因子的产生。DC-SIGN是一种常用的PRR，可以有效地用甘露糖基化抗原靶向以增强抗原特异性T细胞的诱导。天然O-甘露糖苷键对酶促降解敏感，并且其化学敏感性使甘露糖基化抗原的合成复杂化。由于这个原因，（寡）甘露糖苷通常在抗原合成的后期引入，需要正交缀合柄用于其连接。为了增加甘露糖苷的稳定性和简化甘露糖基化肽抗原的合成，我们在这里描述了酸稳定的C-甘露糖基赖氨酸的开发，其允许在固相肽合成（SPPS）期间内联引入甘露糖苷。所开发的氨基酸已成功地用于组装小配体和肽抗原缀合物，所述肽抗原缀合物包含gp 100黑素瘤相关抗原的表位和用于DC活化的TLR 7激动剂。配体表现出类似的内化能力和结合亲和力的O-甘露糖类似物。此外，抗原缀合物能够诱导成熟，刺激促炎细胞因子的分泌，并提供增强的gp 100呈递给CD 8（+）和CD 4（+）T细胞，类似于它们的O-甘露糖基对应物。我们的研究结果表明，C-甘露糖赖氨酸是一个有价值的构建块的抗癌肽缀合物疫苗的形式。
Dendritic cells (DCs) are armed with a multitude of Pattern Recognition Receptors (PRRs) to recognize pathogens and initiate pathogen-tailored T cell responses. In these responses, the maturation of DCs is key, as well as the production of cytokines that help to accomplish T cell responses. DC-SIGN is a frequently exploited PRR that can effectively be targeted with mannosylated antigens to enhance the induction of antigen-specific T cells. The natural O-mannosidic linkage is susceptible to enzymatic degradation, and its chemical sensitivity complicates the synthesis of mannosylated antigens. For this reason, (oligo)mannosides are generally introduced in a late stage of the antigen synthesis, requiring orthogonal conjugation handles for their attachment. To increase the stability of the mannosides and streamline the synthesis of mannosylated peptide antigens, we here describe the development of an acid-stable C-mannosyl lysine, which allows for the inline introduction of mannosides during solid-phase peptide synthesis (SPPS). The developed amino acid has been successfully used for the assembly of both small ligands and peptide antigen conjugates comprising an epitope of the gp100 melanoma-associated antigen and a TLR7 agonist for DC activation. The ligands showed similar internalization capacities and binding affinities as the O-mannosyl analogs. Moreover, the antigen conjugates were capable of inducing maturation, stimulating the secretion of proinflammatory cytokines, and providing enhanced gp100 presentation to CD8(+) and CD4(+) T cells, similar to their O-mannosyl counterparts. Our results demonstrate that the C-mannose lysine is a valuable building block for the generation of anticancer peptide-conjugate vaccine modalities.