Gouyou Baptiste, Millul Jacopo, Villa Alessandra, Cazzamalli Samuele, Neri Dario, Matasci Mattia
Philochem AG, Libernstrasse 3, 8112 Otelfingen, Switzerland.
Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology, 8093 Zurich, Switzerland.
ACS Omega. 2020 Sep 30;5(40):26077-26083. doi: 10.1021/acsomega.0c03592. eCollection 2020 Oct 13.
Small ligands specific to tumor-associated antigens can be used as alternatives to antibodies for the delivery of small payloads such as radionuclides, cytotoxic drugs, and fluorophores. Their use as a delivery moiety of bioactive proteins such as cytokines remains largely unexplored. Here, we describe the preparation and in vivo characterization of the first small molecule-cytokine conjugate targeting carbonic anhydrase IX (CAIX), a marker of renal cell carcinoma and hypoxia. Site-specific conjugation between interleukin-2 and acetazolamide was obtained by sortase A-mediated transpeptidation. Binding of the conjugate to the cognate CAIX antigen was confirmed by surface plasmon resonance. The in vivo targeting of structures expressing carbonic anhydrase IX was assessed by biodistribution experiments in tumor-bearing mice. Optimization of manufacturability and tumor-targeting performance of acetazolamide-cytokine products will be required in order to enable industrial applications.
肿瘤相关抗原特异性的小分子配体可作为抗体的替代品，用于递送放射性核素、细胞毒性药物和荧光团等小分子负载物。它们作为细胞因子等生物活性蛋白的递送部分的应用在很大程度上仍未得到探索。在此，我们描述了首个靶向碳酸酐酶IX（CAIX）的小分子-细胞因子缀合物的制备及其体内表征，CAIX是肾细胞癌和缺氧的标志物。通过分选酶A介导的转肽作用实现了白细胞介素-2与乙酰唑胺之间的位点特异性缀合。通过表面等离子体共振证实了缀合物与同源CAIX抗原的结合。通过在荷瘤小鼠体内进行生物分布实验评估了表达碳酸酐酶IX的结构的体内靶向性。为了实现工业应用，需要优化乙酰唑胺-细胞因子产品的可制造性和肿瘤靶向性能。
