A Novel Immunotolerizing Therapy for Autoimmune Vitiligo

描述（由申请人提供）：Radikal Therapeutics（RTX）是一种新型疗法的先驱，用于恢复白癜风的免疫耐受性和阻止进行性色素脱失。在洛约拉大学（LUC），开发了一种可诱导热休克蛋白70的变体，具有预防和治疗自身免疫性白癜风的显着潜力。发现该变体HSP 70 iQ435 A（命名为“CM”）仅携带对蛋白质的单个氨基酸修饰，具有涉及树突状细胞（DC）的持久耐受化和T细胞流入皮肤的抑制的疗效。使用不同的自发性白癜风T细胞受体转基因小鼠模型，在预防和治疗环境中，CM DNA疫苗接种可显著抑制色素脱失。翻译到人类，炎症性CD11b+CD11c+亚群的DC负责沉淀和永久白癜风中发现增加丰度循环和皮肤浸润DC。因此，我们假设CM同样会干扰人类白癜风的进行性色素脱失。鉴于基础科学研究的实力和创新，以及在多种白癜风动物模型中揭示的深刻疗效，以及临床概念验证的明确途径，我们现在提出SBIR第1阶段建议，通过在临床相关的白癜风大型动物模型中证明疗效来推进CM。具体目标#1：在具有类人皮肤的大型白癜风模型中建立M的功效。在RTX时，将感兴趣的DNA亚克隆到pUMCV 3中，与原始质粒相比，pUMCV 3是一种适合人类使用和表达的载体。然后将在pUMVC 3中产生150 mg CM（>99%纯度），以支持大型动物疗效研究以及后续GLP毒理学和安全药理学研究。我们的合作者Le Poole博士（LUC）将在Sinclair猪中进行研究（每组n=6）。该模型开发白癜风以及黑色素瘤，将用于通过无针喷射注射在模型中测试CM DNA应用，该模型允许在人类皮肤中进行测试，同时测量对抗黑色素瘤反应的副作用。通过量化病变面积的变化来测量疗效。将跟踪局部和全身不良事件，并进行免疫监测，以将早期发现转化为大型动物。这包括分析随时间推移从动物身上采集的血液和皮肤样本中的树突状细胞和T细胞谱。具体目标#2：准备CM用于IND批准。在RTX，将制定临床方案，并计划进行研究以确定NOAEL，最终召开IND前会议，为I期临床研究的IND批准做准备。对于后续GLP毒理学和安全药理学研究，将在pUMVC 3中生成150 mg CM（>99%纯度）。临床研究者将接受CM及其在临床前环境中的应用的培训，并在必要时管理任何不良事件。我们预期，相对于盐水对照组，CM将诱导白癜风进展减少>50%，并且免疫监测将确定T细胞向皮肤的浸润减少> 50%。

DESCRIPTION (provided by applicant): Radikal Therapeutics (RTX) is pioneering a novel therapy to restore immune tolerance and arrest progressive depigmentation in vitiligo. At Loyola University (LUC), a variant to inducible Heat Shock Protein 70 was developed with remarkable potential for the prevention and treatment of autoimmune vitiligo. Carrying only a single amino acid modification to the protein, this variant HSP70iQ435A (designated "CM") was found to have a curative effect involving long-lasting tolerization of dendritic cells (DCs) and inhibition of T ell influx to the skin. Depigmentation was inhibited by CM DNA vaccination to a remarkable extent in both preventive and therapeutic settings using different spontaneous, T cell receptor transgenic mouse models of vitiligo. Translated to humans, the inflammatory CD11b+CD11c+ subset of DCs held responsible for precipitating and perpetuating vitiligo is found in increased abundance among circulating and skin infiltrating DC. We thus hypothesize that the CM will likewise interfere with progressive depigmentation in human vitiligo. Given the strength and innovation of the basic science investigations and the profound efficacy revealed in multiple animal models of vitiligo, and the clear path forward to clinical proof-of-concept, we now propose an SBIR Phase 1 proposal to advance the CM through demonstration of efficacy in a clinically-relevant large animal model of vitiligo. Specific Aim #1: Establish the efficacy of the M in a large vitiligo model with human-like skin. At RTX the DNA of interest will be subcloned into pUMCV3, a vector suited for human use and expression compared to the original plasmid. 150 mg of CM (>99% purity) in pUMVC3 will then be generated to support large animal efficacy studies as well as for follow-on GLP toxicology and safety pharmacology investigations. Our collaborator, Dr. Le Poole (LUC), will carry out studies in Sinclair pigs (n=6 per group). This model, developing vitiligo as well as melanoma, will serve to test CM DNA application by needle-less jet injection in a model that allows for testing in human-like skin while measuring side-effects on anti-melanoma responses. Efficacy is measured by quantifying changes in lesional area. Local and systemic adverse events will be followed, and immune monitoring performed to translate earlier findings to large animals. This includes analysis of dendritic cell and T cell profiles in blood and skin samples taken from the animals over time. Specific Aim #2: Prepare the CM for IND approval. At RTX, a clinical protocol will be prepared and studies planned to determine the NOAEL, culminating in a pre-IND meeting on preparation for IND approval for a Phase I clinical study. For follow-on GLP toxicology and safety pharmacology investigations, 150 mg of CM (>99% purity) in pUMVC3 will be generated. Clinical investigators will be trained in working with the CM and its application in a pre-clinical setting and to manage any adverse events if necessary. We expect that the CM will induce >50% reduction in vitiligo progression relative to the saline control group and that immune monitoring will determine a > 50% reduction in T cell infiltration to the skin.