First-in-class TREM-1 inhibitors for neovascular retinal diseases

项目摘要/摘要 视网膜新生血管(RNV)是早产儿视网膜病变(ROP)视力丧失的主要原因， 糖尿病视网膜病变(DR)和视网膜静脉阻塞。在美国，大约16,000名早产儿是 每年受ROP影响，约410万成人年有Dr.常规并发症 治疗表明，对新疗法的需求尚未得到满足。这个项目的长期目标是开发一种 对ROP和其他RNV疾病的系统、新机制、有效和耐受性好的治疗。 髓样细胞上表达的触发受体(TREM-1)在炎症和IS时上调 参与血管生成信号通路，提示TREM-1有望成为治疗RNV的靶点。 目前的TREM-1抑制剂都试图阻断TREM-1与其仍不确定的配体的结合(S)。至 将临床失败风险降至最低，我们开发了耐受性良好的TREM-1抑制肽GF9和GA31 采用一种新颖的、不依赖配体的作用机制。它们可以自由形式使用，也可以在 巨噬细胞特异性脂肽复合体(LPC)可改善半衰期并降低偏离目标的风险。 此前，我们证明了配体非依赖的TREM-1使用游离(GF9)或LPC-1阻断。 配方多肽(GA31-LPC)：1)防治氧诱导的小鼠视网膜病变的RNV (OIR)；2)改善缺氧时的血管萌发，3)抑制视网膜TREM-1和CSF-1的表达，以及 4)减少细胞因子(肿瘤坏死因子α、白介素1β、白介素6和脑脊液1)的释放，而对照多肽无此作用。 拟议项目的目标是进一步开发这种治疗新生血管的一流的TREM-1疗法。 视网膜疾病。考虑到GF9和GA31-LPC的利弊，我们建议从这两个导联开始。 由于血管表型的表现不同，我们建议同时使用OIR小鼠和大鼠。 第一阶段的目标是：1)生产符合GMP标准的游离GF9和GA31-LPC配方并进行测试 2)在OIR小鼠模型中测试GF9和GA31-LPC的开发配方。 将探索GMP友好的切向流过滤技术来制备GA31-LPC。 第二阶段的目标是：1)建立测定眼组织中GF9和GA31的LC-MS分析方法；2)检测 GF9和GA31-LPC体内药代动力学及眼组织分布 GF9和GA31-LPC对两种OIR模型的治疗作用及导联选择；4)检测导联 非临床毒理学研究。将进行全面的组织学/IHC检查。将对细胞因子进行测试。 后续的IIb阶段将包括其他用药和联合(如激光+GF9)方案，GLP- 毒物、ADME、CMC和其他支持IND的研究。最终产品将代表安全和稳定 系统治疗。其预期的安全性得到了长期治疗的健康人群中GF9治疗的安全性的支持， 癌症和关节炎小鼠。SignaBlok的LPC原型在人类身上耐受性良好。TREM-1封锁 SignaBlok竞争对手的肽LR12(Inotrem)在健康和脓毒症受试者中是安全和耐受性良好的。

Project Summary/Abstract Retinal neovascularization (RNV) is a major cause of vision loss in retinopathy of prematurity (ROP), diabetic retinopathy (DR) and retinal vein occlusion. In the US, about 16,000 of premature infants are affected by ROP annually and about 4.1 million adults years have DR. Complications of conventional treatments suggest an unmet need for new therapies. The long-term objective of this project is to develop a systemic, new mechanism-based, efficient and well-tolerable therapy for ROP and other RNV diseases. Triggering receptor expressed on myeloid cells (TREM-1) is upregulated upon inflammation and is involved in angiogenic signaling pathways, suggesting TREM-1 as a promising target for treatment of RNV. Current TREM-1 inhibitors all attempt to block binding of TREM-1 to its still uncertain ligand(s). To minimize clinical failure risks, we developed well-tolerable TREM-1 inhibitory peptides GF9 and GA31 that employ a novel, ligand-independent mechanism of action. They can be used in a free form or formulated in macrophage-specific lipopeptide complexes (LPC) to improve half-life and reduce off-target risks. Previously, we showed that ligand-independent TREM-1 blockade using either a free (GF9) or LPC- formulated peptide (GA31-LPC): 1) prevents and treats RNV in mice with oxygen-induced retinopathy (OIR); 2) improves vessel sprouting during hypoxia, 3) inhibits retinal TREM-1 and CSF-1 expression, and 4) reduces cytokine release (TNFα, IL-1β, IL-6 and CSF-1) in vitro, while control peptides have no effect. The goal of the proposed project is to further develop this first-in-class TREM-1 therapy for neovascular retinal diseases. Considering pros and cons of GF9 and GA31-LPC, we suggest to start with both leads. Due to differences in the manifestation of vascular phenotypes, we suggest to use both OIR mice and rats. Phase I aims are to: 1) generate GMP-compliant formulations of free GF9 and GA31-LPC and test them in vitro, and 2) test the developed formulations of GF9 and GA31-LPC in the OIR mouse model. GMP-friendly tangential flow filtration technique to prepare GA31-LPC will be explored. Phase II aims are to: 1) develop an LC-MS-assay to measure GF9 and GA31 in ocular tissues, 2) test pharmacokinetics and ocular tissue distribution of GF9 and GA31-LPC in vivo, 3) test the preventative and therapeutic effects of GF9 and GA31-LPC in two OIR models and select the lead, and 4) test the lead in non-clinical toxicology studies. Comprehensive histology/IHC will be performed. Cytokines will be tested. Follow-up Phase IIb will include other administration and combination (eg, laser + GF9) regimens, GLP- TOX, ADME, CMC and other IND-enabling studies. The final product will represent safe and stable systemic therapy. Its anticipated safety is supported by safety of GF9 therapy in long-term treated healthy, cancer and arthritic mice. Prototypes of SignaBlok's LPC are well-tolerated in humans. TREM-1 blockade by SignaBlok competitor's peptide LR12 (Inotrem) is safe and well-tolerated in healthy and septic subjects.