Targeting the eNAMPT/TLR4 pathway to reduce Inflammatory Bowel Disease severity

摘要 炎症性肠病（IBD）是一种使人衰弱的疾病（没有治疗方法），受到严重影响 通过失调的炎症途径。这个 A0 R42 STTR 应用程序专注于高度新颖的人性化 单克隆抗体 (mAb) ALT-100，靶向并中和 eNAMPT（细胞外烟酰胺） 磷酸核糖基转移酶），一种损伤相关分子模式蛋白（DAMP），可强力激活 通过连接 Toll 样受体 4 (TLR4) 来调节先天免疫驱动的炎症通路。我们推测 eNAMPT 中和 ALT-100 mAb 是解决限制性疗法未满足需求的策略 炎症性肠损伤/纤维化并减轻 IBD 的严重程度。已发表和未发表的数据强烈 支持 eNAMPT 参与人类 IBD 病理学。首先，NAMPT RNA和蛋白表达是 IBD 患者肠道组织中的 IBD 显着增加，并且 IBD 相关刺激高度上调 包括生长因子和细胞因子。其次，IBD 受试者血浆/血清 eNAMPT 水平升高 并与疾病严重程度和抗 TNFa 疗法的反应相关。三、NAMPT多态性 （SNP），以前与 ARDS 和肺动脉高压（PH）中炎症性疾病的严重程度有关， GWAS 研究中与 IBD 严重程度相关。最后，令人信服的公布数据表明 eNAMPT 是 高度可成药的靶点，ALT-100 mAb 可以显着减弱多种临床前系统性副作用 炎症/器官损伤（急性呼吸窘迫综合征、肺PH、辐射引起的纤维化、癌症）。重要的是，ALT-100 mAb 降低临床前 IBD 严重程度、结肠炎症和纤维化。 ALT-100 mAb 作为抗体的可行性 IBD 治疗得到已完成的急性 IND 药代动力学 (PK) 研究和 28 天毒性研究的支持 研究显示 IV ALT-100 mAb 制剂的 T1/2 半衰期为 10 天，并且没有明显的毒性 （老鼠和猪）。此外，我们还完成了CMC制造开发（稳定细胞系、 研究/主细胞库）和 IV ALT-1000 mAb 的 200L GMP 生物反应器运行（表达 6 gms/L） 配方（10毫克/毫升）。我们预计 FDA 将于 2022 年 5 月提交 ARDS 的 IND 申请。在 R-42 的第一阶段 STTR 阶段 I/II 快速通道申请，具体目标 #1/2 将评估药效 (PD) 特性 ALT-100 mAb 用于治疗已证实的急性（7 天）和慢性（28 天）临床前 DSS 诱导的小鼠结肠炎 建立模型并优化 ALT-100 mAb 剂量（0.4 mg/kg、1mg/kg、4 mg/kg）和给药途径（IV 与 subQ） 使用 IV 制剂 (10 mg/mL) 和 subQ 制剂 (100 mg/mL)。在大鼠中进行的 II 期研究 和小型猪，具体目标 #3 将描述 subQ 和 IV 的药代动力学 (PK) 特征 ALT-100 mAb 配方和特定目标 #4 将评估慢性 subQ ALT-100 mAb 毒代动力学 属性。这些 I/II 期 STTR 研究的成功完成将确立 ALT-100 mAb 的可行性 IBD 治疗并能够提交 FDA IND 申请，以允许 Aqualung 进行 IBD 临床 将 ALT-100 mAb 作为一种新型疗法来解决重症患者未满足的需求的试验 IBD，尤其是生物疗法失败的受试者。

ABSTRACT Inflammatory bowel disease (IBD) is a debilitating disease (without curative therapies) that is critically influenced by dysregulated inflammatory pathways. This A0 R42 STTR application focuses on a highly novel humanized monoclonal antibody (mAb), ALT-100, to target and neutralize eNAMPT (extracellular nicotinamide phosphoribosyltransferase), a damage-associated molecular pattern protein (DAMP) that robustly activates innate immunity-driven inflammatory pathways via ligation of the Toll-like receptor 4 (TLR4). We speculate the eNAMPT-neutralizing ALT-100 mAb to be a strategy to address the unmet need for therapies that limit inflammatory bowel injury/fibrosis and reduce the severity of IBD. Published and unpublished data strongly support involvement of eNAMPT in human IBD pathobiology. First, NAMPT RNA and protein expression are significantly increased in intestinal tissues from IBD patients and are highly upregulated by IBD-relevant stimuli including growth factors and cytokines. Second, plasma/serum eNAMPT levels are elevated in IBD subjects and correlate with disease severity and responses to anti-TNFa therapies. Thirdly, NAMPT polymorphisms (SNPs), previously linked to inflammatory disease severity in ARDS and pulmonary hypertension (PH), associate with IBD severity in GWAS studies. Lastly, compellingly published data demonstrate eNAMPT is a highly druggable target, with ALT-100 mAb profoundly attenuating multiple preclinical systemic inflammation/organ injuries (ARDS, PH, radiation-induced fibrosis, cancer). Importantly, ALT-100 mAb reduces preclinical IBD severity, colon inflammation and fibrosis. The feasibility of ALT-100 mAb as an IBD therapy is supported by completed acute IND-enabling pharmacokinetic (PK) studies and 28-day toxicity studies showing the IV ALT-100 mAb formulation has a T1/2 half-life of 10 days and is without discernable toxicity (rats and pigs). In addition, we have completed CMC manufacturing development (stable cell line, Research/Master Cell Banks) and a 200L GMP Bioreactor run (expression 6 gms/L) of the IV ALT-1000 mAb formulation (10mg/mL). We anticipate FDA IND submission for ARDS in May 2022. In PHASE I of this R-42 STTR PHASE I/II Fast Track application, Specific Aims #1/2 will assess pharmacodynamic (PD) properties of ALT-100 mAb in well-established acute (7 days) and chronic (28 days) preclinical DSS-induced murine colitis models and optimize ALT-100 mAb dosing (0.4 mg/kg, 1mg/kg, 4 mg/kg) and route of delivery (IV vs subQ) using IV formulations (10 mg/mL) and subQ formulations (100 mg/mL). In PHASE II studies conducted in rats and minipigs, Specific Aim #3 will characterize the pharmacokinetic (PK) characteristics of the subQ and IV ALT-100 mAb formulations and Specific Aim #4 will evaluate chronic subQ ALT-100 mAb toxicokinetic properties. Successful completion of these PHASE I/ II STTR studies will establish ALT-100 mAb as a viable IBD therapeutic and enable the submission of an FDA IND application to allow Aqualung to conduct IBD clinical trials that address ALT-100 mAb as a novel therapeutic to address the unmet needs in subjects with severe IBD, especially in subjects who fail biologic therapies.