A Novel Probiotic for the Treatment of Sjogren's Syndrome

项目摘要 我们的目标是开发一种新型的，免疫增强的乳酸乳杆菌益生菌为基础的治疗 干燥综合征(SjS)的治疗。SJS是一种进行性慢性自身免疫性疾病，其特征是 炎性细胞渗入唾液和泪腺，导致腺泡上皮细胞萎缩。 死亡，外分泌功能丧失1-6。至少一半的SjS患者发展为腺外炎症 并有广泛的全身性临床表现，可影响任何器官系统，包括 结缔组织，5-10%的患者患上危及生命的淋巴瘤7、8。SJS是一种衰弱的疾病 在美国，多达310万人受到影响。9，10，其中女性患病的可能性是女性的9倍 患有Sjs的人比男性多5、10、11岁。 SjS的治疗仍然是一个重要的未得到满足的医疗需求。目前的治疗依赖于替代疗法 例如人工唾液和眼部润滑剂或免疫抑制剂12、13。因为多种抗原 α-fodrin 14-17、核糖核蛋白Ro/SSA14、48、49、La/SSB14、48、49和 M3R18，49，50，口服耐受性方法变得有问题。因此，刺激调节细胞的能力 与已知该病的抗原特异性无关，它被认为是一种有吸引力的治疗方法。 最初被认为是人类腹泻疫苗，我们发现定居因子抗原I(CFA/I)来自 人肠毒素大肠杆菌(ETEC)在以下情况下可有效诱导自身抗原特异性T调节细胞 口服作为纯化蛋白或通过沙门氏菌或乳酸乳杆菌细菌传递 19-21，61号系统。为了避免与大量生产CFA/I蛋白相关的挑战，并改善 CFA/I口服后的粘膜药代动力学(PK)和药效学(PD)特性 我们开发并鉴定了含有乳酸乳杆菌CFA/I的载体表达产物(简称VTC-I)。 CFA)，并在多种自身免疫模型中评估其疗效。事实上，职训局-终审法院在减少 SjS动物模型的临床症状，以及RA、糖尿病和MS 22-24的实验模型。之前 第一阶段和第二阶段SBIR的资助使我们能够有效地推动VTC-CFA走向临床测试，方法是 开发从基因组整合操纵子表达CFA/I的临床候选，验证非 临床生物学，并完成上下游流程开发。作为这些努力的一部分，我们 召开FDA Pre-IND会议，最终确定我们的IND支持计划，并调整我们的CMC/制造战略。 此申请旨在完成VTC-CFA IND启用研究，并向FDA提交IND。钥匙 这项建议的目的是：1)完成VTC-CFA GLP毒理学研究，2)建立cGMP生产 基础设施，3)进行cGMP药材制造，4)在cGMP条件下灌装胶囊 药品，以及5)向FDA提交VTC-CFA的IND。 VTC-CFA的成功商业化将为SJS的治疗提供深刻的医学进步。

Project Summary Our goal is to develop a novel, immunologically enhanced L. lactis probiotic-based therapeutic for the treatment of Sjögren’s Syndrome (SjS). SjS is a progressive, chronic autoimmune disease characterized by inflammatory cell infiltration of the salivary and lacrimal glands, resulting in acinar epithelial cell atrophy, cell death, and loss of exocrine function 1-6. At least half of SjS patients develop extraglandular inflammatory disease and have a wide range of systemic clinical manifestations that can affect any organ system, including connective tissue, and 5-10% of patients develop life-threatening lymphoma 7, 8. SjS is a debilitating disease affecting as many as 3.1 million individuals in the U.S. 9, 10, with women being nine times more likely to be afflicted with SjS than men 5, 10, 11. Treatment of SjS remains a significant unmet medical need. Current treatment relies on replacement therapies such as artificial saliva and eye lubricants or immunosuppressive agents12, 13. Because of the multiple antigens involved in this disease process, i.e., α-fodrin 14-17, ribonuclear protein Ro/SSA 14, 48, 49, La/SSB 14, 48, 49, and M3R 18, 49, 50, oral tolerance methods become problematic. Thus, the capacity to stimulate regulatory cells independent of knowing the antigen specificity for the disease poses as an attractive therapeutic approach. Originally conceived as a human diarrheal vaccine, we found that colonization factor antigen I (CFA/I) from human enterotoxigenic E. coli (ETEC) is effective at inducing auto-Ag-specific T regulatory cells when administered orally as either a purified protein or delivered via a Salmonella or L. lactis bacterial delivery system 19-21, 61. To avoid challenges associated with producing large quantities of CFA/I protein and improve the mucosal pharmacokinetic (PK) and pharmacodynamic (PD) properties of CFA/I following oral administration, we developed and characterized a vector-containing L. lactis-CFA/I expressing product (referred to as VTC- CFA) and evaluated its efficacy in multiple autoimmune models. Indeed, VTC-CFA was effective at reducing clinical symptoms in a SjS animal model, along with experimental models of RA, diabetes, and MS 22-24. Prior Phase I and II SBIR funding has enabled us to effectively advance VTC-CFA towards clinical testing by developing our clinical candidate that expresses CFA/I from a genome-integrated operon, validating non- clinical biology, and completing upstream and downstream process development. As part of these efforts, we held an FDA pre-IND meeting to finalize our IND-enabling plans and align our CMC/manufacturing strategy. This application is designed to complete VTC-CFA IND enabling studies and file an IND with the FDA. The key aims of this proposal are: 1) complete VTC-CFA GLP toxicology studies, 2) establish cGMP manufacturing infrastructure, 3) perform cGMP manufacturing for drug substance, 4) Fill capsules under cGMP conditions for drug product, and 5) file an IND with the FDA for VTC-CFA. Successful commercialization of VTC-CFA will provide a profound medical advancement for treating SjS.