Development of an engineered CCL20 protein as a lead therapeutic molecule for psoriasis

项目总结/摘要 该项目的目标是开发和验证一种新的治疗先导化合物，用于治疗 银屑病趋化因子在正常免疫应答期间协调炎性细胞的迁移， 是免疫组织发育和体内平衡所必需的。当异常的趋化因子功能发生时， 免疫细胞的不适当募集可导致具有破坏性影响的多种炎性病理 病人的生活质量趋化因子CCL 20及其G蛋白偶联受体CCR 6驱动细胞凋亡。 通过启动和持续募集表达炎性Th 17的 细胞进入真皮。我们最近发表的生物化学，细胞基础和体内研究证明， 完全模拟天然CCL 20分子二聚体形式的工程重组蛋白 逆转其正常的促炎功能。这种先导化合物（CCL 20 LD）由PI发明， 威斯康星州医学院，防止银屑病皮炎（PsD）的发展和进展， 一种忠实地再现人类银屑病的临床前小鼠模型。在PF的SBIR第一阶段赠款授予期间 在此期间，我们成功完成了CCL 20 LD临床前开发的所有三个里程碑， 牛皮癣疗法首先，使用一种新的系统性IL-23微环模型，Protein Foundry表明CCL 20 LD 减轻疾病建立后的皮肤炎症症状，证明CCL 20 LD是有效的 作为临床相关的人银屑病模型的治疗性治疗。第二，CCL 20 LD是 在初步血清学分析中显示相对安全，没有肝或肾病累积， 组织损伤的生物标志物。第三，PF验证了可重现的生产和质量控制方案， CCL 20 LD蛋白。鉴于第一阶段工作的成功完成和在动物中的显著疗效， 我们建议开发CCL 20 LD作为银屑病的下一代治疗方法。进行R/R&D IND所必需的，该II期申请建议使用以下方法开发优化的给药方案： 临床相关动物模型，并验证啮齿动物的安全性特征，以指导IND使能GLP 非人类灵长类动物的毒理学研究。在本申请的目的1中，Protein Foundry及其学术研究 加州大学戴维斯分校的合作者将建立治疗剂量和时间表，产生最强的抗- 在最短的时间内达到治疗银屑病的效果。在目标2中，Protein Foundry将使用细胞因子风暴和抗药物 抗体试验，用于检测免疫原性并在啮齿动物中建立毒理学特征，以指导GLP毒理学 非人类灵长类动物的研究。最后，在目标3中，Protein Foundry将探索用于测试的局部制剂， 动物模型作为区分CCL 20 LD与当前市场治疗的第一个关键步骤， 建立CCL 20 LD的目标产品档案。通过其天然的基因工程改变CCR 6信号传导 配体是用于治疗自身炎性疾病的临床方法的创新范式转变。 开发工程化的CCL 20变体作为生物治疗剂将具有显著的积极影响 通过减少副作用和提供具有延长的治疗寿命的药物来治疗银屑病患者。 此外，所得方法具有治疗其它Th 17介导的疾病的潜力。

Project Summary/Abstract The goal of this project is to develop and validate a novel therapeutic lead compound for the treatment of psoriasis. Chemokines orchestrate the migration of inflammatory cells during normal immune responses and are required for immune tissue development and homeostasis. When aberrant chemokine function occurs, improper recruitment of immune cells can lead to a variety of inflammatory pathologies with devastating effects on a patient’s quality of life. The chemokine CCL20 and its G protein-coupled receptor CCR6 drive the development of psoriasis through the initiation and continuous recruitment of inflammatory Th17-expressing cells into the dermis. Our recently published biochemical, cell-based and in vivo studies prove that an engineered recombinant protein that mimics the dimeric version of the natural CCL20 molecule completely reverses its normal pro-inflammatory functional profile. This lead compound (CCL20LD), invented by the PI at the Medical College of Wisconsin, prevented the development and progression of psoriatic dermatitis (PsD) in a preclinical mouse model that faithfully recapitulates human psoriasis. During PF’s SBIR Phase I grant award period, we successfully completed all three milestones focused on the preclinical development of CCL20LD as a psoriatic therapy. First, using a new systemic, IL-23 minicircle model, Protein Foundry showed that CCL20LD alleviated signs of skin inflammation after disease establishment, demonstrating that CCL20LD is efficacious as a therapeutic treatment in a clinically relevant model of human psoriasis. Second, CCL20LD was shown in preliminary serological analyses to be relatively safe, with no accumulation of hepatic or nephrotic biomarkers of tissue damage. Third, PF validated a reproducible manufacturing and quality control protocol for the CCL20LD protein. Given the successful completion of the phase I work and the notable efficacy in animals, we propose to develop CCL20LD as a next-generation treatment for psoriasis. To perform the R/R&D necessary for an IND, this phase II application proposes to develop an optimized dosing schedule using clinically relevant animal models, and validate the safety profile in rodents to guide IND-enabling GLP toxicology studies in non-human primates. In Aim 1 of this application, Protein Foundry and its academic collaborators at UC Davis will establish the therapeutic dose and schedule which produces the strongest anti- psoriatic effect in the shortest time period. In Aim 2, Protein Foundry will use cytokine storm and anti-drug antibody assays to test immunogenicity and establish a toxicology profile in rodents to guide GLP toxicology studies in non-human primates. Lastly, in Aim 3, Protein Foundry will explore topical formulations for testing in animal models as a first key step in differentiating CCL20LD from current market therapeutics and fully establish CCL20LD’s targeted product profile. Altering CCR6 signaling through the engineering of its native ligand is an innovative paradigm shift in clinical approaches for treating auto-inflammatory diseases. Development of engineered CCL20 variants as biological therapeutics will have significant positive impact for psoriasis patients by reducing side effects and providing a drug with extended therapeutic lifetimes. Moreover, the resultant method has the potential for treating other Th17-mediated diseases.