A novel immunotolerizing therapy for autoimmune vitiligo

Radikal Treeutics(RTX)正在开发一种恢复免疫耐受和停滞的开创性疗法 白癜风的进行性色素脱失。在我们的合作机构，洛约拉大学芝加哥分校， 诱导型热休克蛋白70在预防和治疗方面具有显著的潜力 自身免疫性白癜风。这种变种只对蛋白质进行单一的氨基酸修饰 HSP70iQ435A(“CM”)被发现具有与树突状细胞长期耐受有关的疗效 (DCs)和抑制T细胞进入皮肤。在第1阶段SBIR中，我们检查了 辛克莱猪的自发性色素脱失，其特征是黑色素瘤退行性和新的 发展中的白癜风。与临床表现相平行，在该系统中炎性CD11b+CD11c+ 被认为对白癜风的形成和延续负有责任的DC亚群越来越多 在循环和皮肤浸润性DC之间。我们观察到对照组中未治疗的皮损 逐渐增大的大小增加了32%，而在CM治疗的皮损中观察到37%的重新着色 (P=0.0045)。这种皮肤色素沉着的变化与处理后的猪减少了50%有关 浸润性T细胞(P&lt；0.04)。因此，我们假设编码CM的DNA同样会干扰 人类白癜风患者进行性色素脱失，为CM的发展提供了诱因 变成一种适销对路的药物。目标1：放大生产GMP级CM PI将合成新产品 高效表达CM质粒的大肠杆菌克隆。腾讯通将最终确定增长的优化 条件，开发特定产品的高效液相色谱释放分析，并产生一个参考标准。阿尔德夫龙将 生成表达CM的主细胞库和工作细胞库，并生成g GMP批次的 CM，以支持白癜风临床1a期安全性和有效性研究，并进行 稳定性分析。将开发分析方法来表征CM的GMP释放和稳定性 学习。目的#2：在白癜风小鼠模型中，CM的治疗效果与病程有关。腾讯通将 测量中药对进行性白癜风h3TA2小鼠的疗效与病程的关系。我们会 对发病6-36周的小鼠进行治疗，通过扫描来测量疾病的停止和重新着色。目标3： 在FDA IND要求的GLP毒理学研究中确定CM的急性安全性、毒性和耐受性 申请。RTX将进行一项为期13周的GLP研究，其中CM通过皮下途径给药 为了阐明NOAEL在小鼠体内的作用并为测试的剂量范围提供依据 人类的安全和宽容。目标4：编写和准备FDA RTX将提交的IND前申请 准备并提交法规文件，以支持临床GCP阶段1a研究，以评估 CM在活动期白癜风志愿者中的安全性。RTX将与FDA会面，介绍我们的 疗效数据在2022年导致药物注册的临床注册路径上获得一致。

Radikal Therapeutics (RTX) is developing a pioneering therapy to restore immunotolerance and arrest progressive depigmentation in vitiligo. At our partnering institution, Loyola University Chicago, 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 (“CM”) was found to have a curative effect involving long-lasting tolerization of dendritic cells (DCs) and inhibition of T cell influx to the skin. In the Phase 1 SBIR we examined the CM in a model of spontaneous depigmentation in Sinclair swine, characterized by regressing melanoma and newly developing vitiligo. Paralleling the clinical presentation, in this system 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 observed that untreated lesions in the control group gradually increased in size by 32%, whereas repigmentation of 37% was observed in CM treated lesions (p=0.0045). This change in cutaneous pigmentation was associated in treated pigs with a 50% reduction in infiltrating T cells (p<0.04). We thus hypothesize that the CM-encoding DNA will likewise interfere with progressive depigmentation in human vitiligo patients, providing incentive for the development of the CM into a marketable drug. Aim #1: Scale-up and produce GMP-grade CM The PI will synthesize de novo a high-producing E. coli clone expressing the CM plasmid. RTX will finalize optimization of the growth conditions, develop product-specific HPLC release assays, and generate a reference standard. Aldevron will generate a Master Cell Bank and Working Cell Bank expressing the CM, and generate a g GMP batch of the CM in order to support clinical Phase 1a safety and efficacy investigations in clinical vitiligo, and to perform stability analysis. Analytical methods will be developed to characterize the CM for GMP release and stability studies. Aim #2: Relate CM treatment efficacy to disease duration in a murine model of vitiligo. RTX will measure efficacy of the CM in relation to disease duration in h3TA2 mice with progressive vitiligo. We will treat mice 6-36 weeks at age at onset to measure disease arrest and repigmentiation by scanning. Aim #3: Establish the acute safety, toxicity, and tolerance of CM in GLP toxicology studies required for FDA IND application. RTX will carry out a 13-week GLP study wherein the CM is dosed via a subcutaneous route of administration in order to elucidate the NOAEL in mice and provide the basis for the dose range to test for safety and tolerance in man. Aim #4: Compile and prepare a pre-IND application to the FDA RTX will prepare and submit regulatory documentation to support a clinical GCP Phase 1a study to evaluate the safety of the CM in human volunteers with active vitiligo. RTX will meet with the FDA to present our efficacy data to gain concurrence on a clinical registration pathway leading to drug registration in 2022.