TREM-1 therapy for rheumatoid arthritis

项目总结/摘要 风湿性关节炎（RA）是一种慢性全身性炎症性疾病， 关节炎影响了大约150万美国人，每年给社会造成400多亿美元的损失。尽管 随着治疗的进展，RA无法治愈，30-40%的RA患者对一线治疗无反应 RA -甲氨蝶呤（MTX），强调需要新的治疗方法。该项目的长期目标是 开发一种新的，一流的，有效的和良好的耐受性治疗RA。 滑膜巨噬细胞上TREM-1的过表达以及TREM-1的丰度和活化， RA患者滑膜中的巨噬细胞与RA的严重程度相关。目前的TREM-1抑制剂 所有这些都试图阻断不确定配体与TREM-1的结合。为了最大限度地降低临床失败风险，我们 开发了配体非依赖性TREM-1抑制肽GF 9、GA 31和GE 31， 转化为巨噬细胞特异性脂肽复合物（LPC），以改善半衰期并降低脱靶风险。 以前，我们表明：1）在CIA小鼠中，GF 9治疗减少了全身炎症并抑制了 与0.1 mg/kg地塞米松一样有效，广泛用于临床治疗RA; 2）GF 9 在体外和体内，治疗可减少细胞因子释放（TNFα、IL-1β、IL-6和CSF-1）达5倍， 对照肽没有影响; 3）GF 9、GF 9-LPC和GA/E31-LPC是无毒的并且被小鼠良好耐受 至少高达300 mg GF 9/kg;和4）将GF 9配制到LPC中使有效剂量减少80%。 该项目的目标是进一步开发这种一流的耐受性良好的TREM-1疗法 单独或与MTX作为诱导/维持治疗。将检测全身和经口给药。 第一阶段（第一年）的目标是：1）制备和测试GMP友好的制剂的自由和LPC结合 体外三功能肽GA 31和GE 31，和2）测试游离的和LPC结合的三功能肽GA 31 和GE 31在CIA小鼠模型中的作用。将使用GMP友好的微流体方法来制备肽-LPC。 第二阶段（第2年和第3年）的目标是：1）开发一种LC-MS测定法来测量血液中的GA 31和GE 31，2） 测定游离和LPC结合的肽GA 31和GE 31的药代动力学，并选择两个先导物，3）测试 目标2中确定的两种导联在两种类风湿动物模型中作为单药治疗和与甲氨蝶呤联合治疗 关节炎，以及4）在毒理学研究中进一步测试这些电极导线，并选择IIb期的电极导线候选者。 IIb期随访将包括测试其他给药（例如，透皮）和联合给药（例如， 泼尼松）方案，进一步优化了先导物及其制造工艺，TOX、ADME、CMC 和其他IND赋能研究。完成后，将提交IND申请。最终产品将 代表安全稳定的RA治疗。其预期的安全性得到了GF 9治疗长期安全性的支持。 治疗健康和关节炎小鼠。SignaBlok的LPC原型在人类中耐受性良好。TREM-1 SignaBlok竞争肽LR 12的阻断在健康和脓毒症受试者中是安全和耐受良好的。

Project Summary/Abstract Rheumatoid arthritis (RA), a chronic, systemic inflammatory disorder that causes chronic inflammation of the joints, affects about 1.5 million Americans and costs society more than $40 billion each year. Despite advances in therapy, RA has no cure and 30-40% of RA patients do not respond to the first-line treatment of RA – methotrexate (MTX), highlighting the need for new treatments. The long-term goal of this project is to develop a novel, first-in-class, efficient and well-tolerable therapy for RA. Overexpression of TREM-1 on the synovial macrophages as well as the abundance and activation of macrophages in the synovium of RA patients correlate with the severity of RA. Current TREM-1 inhibitors all attempt to block binding of the uncertain ligand(s) to TREM-1. To minimize clinical failure risks, we developed a ligand-independent TREM-1 inhibitory peptides GF9, GA31 and GE31 that can be formulated into macrophage-specific lipopeptide complexes (LPC) to improve half-life and reduce off-target risks. Previously, we showed that: 1) in CIA mice, GF9 therapy reduces systemic inflammation and inhibits arthritis as effective as 0.1 mg/kg dexamethasone, widely used in the clinical treatment of RA; 2) GF9 therapy reduces cytokine release (TNFα, IL-1β, IL-6 and CSF-1) in vitro and in vivo up to 5-fold, while control peptide has no effect; 3) GF9, GF9-LPC and GA/E31-LPC are non-toxic and well-tolerated by mice at least up to 300 mg GF9/kg; and 4) formulation of GF9 into LPC reduces the effective dose by 80%. The goal of this project is to further develop this first-in-class well tolerable TREM-1 therapy to be used standalone or with MTX as induction/maintenance therapy. Systemic and oral delivery will be tested. Phase I (Year 1) aims are to: 1) prepare and test GMP-friendly formulations of free and LPC-bound trifunctional peptides GA31 and GE31 in vitro, and 2) test free and LPC-bound trifunctional peptides GA31 and GE31 in the CIA mouse model. GMP-friendly microfluidic method to prepare peptide-LPC will be used. Phase II (Years 2 & 3) aims are to: 1) develop an LC-MS assay to measure GA31 and GE31 in blood, 2) determine pharmacokinetics of free and LPC-bound peptides GA31 and GE31 and select two leads, 3) test two leads identified in Aim 2 as monotherapy and with methotrexate in two animal models of rheumatoid arthritis, and 4) test the leads further in toxicology studies and select the lead candidate for Phase IIb. Follow-up Phase IIb will include testing other administration (eg, transdermal) and combination (eg, Prednisone) regimen, further optimization of the lead and its manufacturing technology, TOX, ADME, CMC and other IND-enabling studies. Upon completion, an IND application will be filed. The final product will represent safe and stable RA therapy. Its anticipated safety is supported by safety of GF9 therapy in long treated healthy and arthritic mice. SignaBlok's LPC prototypes are well tolerated in humans. TREM-1 blockade by SignaBlok competitor's peptide LR12 is safe and well-tolerated in healthy and septic subjects.