Globo H, a specific carbohydrate antigen overexpressed on various human malignancies, has attracted considerable interest as an antigenic target for anticancer vaccine development. Despite several Globo H‐based carbohydrate vaccines that have been designed, efficient access to Globo H hexasaccharide antigen and development of powerful adjuvants for enhancing antitumor immunity remain challenging. Herein, we reported a streamlined chemoenzymatic approach to prepare this hexasaccharide antigen, relying on chemical synthesis of Gb5 pentasaccharide by a stereoconvergent [2+3] strategy and subsequent enzymatic α‐fucosylation to easily install α1,2‐fucose residue. Separately, a modular assembly approach to efficiently synthesize 3‐O‐deacyl‐monophosphoryl lipid A (3D‐MPL) was developed by the integration of stereocontrolled glycosylation, regioselective acylation, site‐specific phosphorylation, and facile global deprotection. After efficient construction of Globo H‐CRM197 conjugate, we conducted systematic immunological evaluations of Globo H‐CRM197 formulated with various adjuvants and adjuvant combinations, comprising saponin QS‐21, synthetic 3D‐MPL and α‐galactosylceramide derivative S34. The results revealed that Globo H‐CRM197 conjugate adjuvanted with QS‐21 and 3D‐MPL elicited robust IgG2a and IgG3 antibody responses and Th1 cellular immunity in mice. Moreover, antibodies induced by this formulation effectively bound to Globo H‐positive MCF‐7 cancer cells and exhibited superior complement‐dependent cytotoxicity and antibody‐dependent cellular phagocytosis, holding promise for further development of effective anticancer vaccines.

中文翻译：

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通过 3D-MPL 和 QS-21 的组合佐剂增强的基于 Globo H 的疫苗的抗肿瘤免疫力


Globo H 是一种在各种人类恶性肿瘤上过表达的特异性碳水化合物抗原，作为抗癌疫苗开发的抗原靶点引起了相当大的兴趣。尽管已经设计了几种基于 Globo H 的碳水化合物疫苗，但有效获取 Globo H 六糖抗原和开发用于增强抗肿瘤免疫力的强效佐剂仍然具有挑战性。在此，我们报道了一种简化的化学酶法来制备这种六糖抗原，该方法依赖于通过立体收敛 [2+3] 策略化学合成 Gb5 五糖，随后的酶促 α-岩藻糖基化以轻松安装 α1,2-岩藻糖残基。另外，通过整合立体控制糖基化、区域选择性酰化、位点特异性磷酸化和简单的全局脱保护，开发了一种有效合成 3-O-十酰基-单磷酸酯脂质 A （3D-MPL） 的模块化组装方法。在高效构建 Globo H-CRM197 偶联物后，我们对由各种佐剂和佐剂组合配制的 Globo H-CRM197 进行了系统的免疫学评估，包括皂苷 QS-21、合成 3D-MPL 和 α-半乳糖神经酰胺衍生物 S34。结果显示，Globo H-CRM197 偶联物辅以 QS-21 和 3D-MPL 在小鼠中引发了强大的 IgG2a 和 IgG3 抗体反应以及 Th1 细胞免疫。此外，由该制剂诱导的抗体与 Globo H 阳性 MCF-7 癌细胞有效结合，并表现出优异的补体依赖性细胞毒性和抗体依赖性细胞吞噬作用，有望进一步开发有效的抗癌疫苗。