Key Points Human IL-10 genetically fused to Chx exotoxin transcytosis domain (AMT-101). AMT-101 was tested in human cell models in vitro and in rodents and nonhuman primates in vivo. Oral dosing targeted intestinal lamina propria to suppress inflammation without systemic PK. IL-10 is a potent anti-inflammatory cytokine capable of suppressing a number of proinflammatory signals associated with intestinal inflammatory diseases, such as ulcerative colitis and Crohn's disease. Clinical use of human IL-10 (hIL-10) has been limited by anemia and thrombocytopenia following systemic injection, side effects that might be eliminated by a gut-restricted distribution. We have identified a transcytosis pathway used by cholix, an exotoxin secreted by nonpandemic forms of the intestinal pathogen Vibrio cholerae. A nontoxic fragment of the first 386 aa of cholix was genetically fused to hIL-10 to produce recombinant AMT-101. In vitro and in vivo characterization of AMT-101 showed it to efficiently cross healthy human intestinal epithelium (SMI-100) by a vesicular transcytosis process, activate hIL-10 receptors in an engineered U2OS osteosarcoma cell line, and increase cellular phospho-STAT3 levels in J774.2 mouse macrophage cells. AMT-101 was taken up by inflamed intestinal mucosa and activated pSTAT3 in the lamina propria with limited systemic distribution. AMT-101 administered to healthy mice by oral gavage or to cynomolgus monkeys (nonhuman primates) by colonic spray increased circulating levels of IL-1R antagonist (IL-1Ra). Oral gavage of AMT-101 in two mouse models of induced colitis prevented associated pathological events and plasma cytokine changes. Overall, these studies suggest that AMT-101 can efficiently overcome the epithelial barrier to focus biologically active IL-10 to the intestinal lamina propria.

中文翻译：

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IL-10 的新型融合，设计为穿越肠道上皮以治疗结肠炎

关键点 人 IL-10 与 Chx 外毒素转胞吞结构域 (AMT-101) 基因融合。AMT-101 在体外人类细胞模型和啮齿动物和非人类灵长类动物体内进行了测试。口服给药靶向肠固有层以抑制炎症而没有全身 PK。IL-10 是一种有效的抗炎细胞因子，能够抑制许多与肠道炎症疾病（如溃疡性结肠炎和克罗恩病）相关的促炎信号。人体 IL-10 (hIL-10) 的临床应用受到全身注射后贫血和血小板减少症的限制，这些副作用可能通过肠道限制分布而消除。我们已经确定了 cholix 使用的转胞吞通路，这是一种由肠道病原体霍乱弧菌的非大流行形式分泌的外毒素。cholix 前 386 个氨基酸的无毒片段与 hIL-10 基因融合以产生重组 AMT-101。AMT-101 的体外和体内表征表明，它可以通过囊泡转胞作用过程有效穿过健康的人肠上皮 (SMI-100)，激活工程化 U2OS 骨肉瘤细胞系中的 hIL-10 受体，并增加细胞磷酸化 STAT3 水平在 J774.2 小鼠巨噬细胞中。AMT-101 被发炎的肠粘膜吸收，并在全身分布有限的固有层中激活 pSTAT3。通过灌胃给健康小鼠或通过结肠喷雾给食蟹猴（非人类灵长类）的 AMT-101 增加了 IL-1R 拮抗剂 (IL-1Ra) 的循环水平。在两种诱导性结肠炎小鼠模型中口服灌胃 AMT-101 可预防相关的病理事件和血浆细胞因子的变化。总的来说，这些研究表明 AMT-101 可以有效地克服上皮屏障，将生物活性 IL-10 集中到肠固有层。