针对肿瘤微环境的抗体可用于传递促炎有效负载，例如细胞因子。细胞因子是能够调节免疫系统活性的小蛋白质，抗体-细胞因子融合蛋白已在临床前和临床环境中进行了测试。在这项研究中，我们描述了 Tripokin，一种新型多特异性融合蛋白，结合了白细胞介素 2 和肿瘤坏死因子的单个氨基酸突变体。这两种促炎有效负载与 L19 抗体融合，L19 抗体是一种针对纤连蛋白外域 B 的临床级抗体。人类有效负载用于临床应用，而相应的鼠细胞因子用于临床前研究。所得融合蛋白在哺乳动物细胞中产生并纯化至均质。鼠类 Tripokin 产品在荷瘤小鼠中具有良好的耐受性，每隔 2 天服用 3 个 30μg 剂量，并在鼠类模型中促进肿瘤快速根除，比单剂免疫细胞因子更有效。 Tripokin 诱导肿瘤快速坏死并刺激强大的免疫反应，影响先天性和适应性免疫途径。此外，与免疫检查点抑制剂的组合进一步增强了我们分子的治疗功效。 Tripokin 代表了一种有前途的临床候选药物，可将白细胞介素 2 和肿瘤坏死因子同时递送至肿瘤部位。© 2024 UICC。

Antibodies that target the tumor microenvironment can be used to deliver pro-inflammatory payloads, such as cytokines. Cytokines are small proteins able to modulate the activity of the immune system, and antibody-cytokine fusion proteins have been tested in preclinical and clinical settings. In this study, we describe Tripokin, a novel multi-specific fusion protein that combines interleukin-2 and a single amino acid mutant of tumor necrosis factor. The two pro-inflammatory payloads were fused to the L19 antibody, a clinical-grade antibody against the extradomain B of fibronectin. The human payloads were used for clinical applications, while the corresponding murine cytokines were used for preclinical studies. The resulting fusion proteins were produced in mammalian cells and purified to homogeneity. The murine Tripokin product was well tolerated in tumor-bearing mice at three doses of 30 μg in a 2-day interval and promoted rapid tumor eradication in murine models, more efficiently than single-agent immunocytokines. Tripokin induced rapid tumor necrosis and stimulated a robust immune response, impacting innate and adaptive immune pathways. In addition, the combination with immune checkpoint inhibitors further boosted the therapeutic efficacy of our molecule. Tripokin represents a promising clinical candidate for the simultaneous delivery of interleukin-2 and tumor necrosis factor to neoplastic sites.© 2024 UICC.