A Small Molecule Enzyme Replacement for the Treatment of CLN1

项目总结 意义：CLN1是一种毁灭性的神经退行性疾病，全世界出生的10万名儿童中就有1名受到影响。 这种无法治愈的疾病的特点是发育急剧下降，神经肌肉症状，视力 丧失和神经元死亡。不幸的是，这种疾病的患者很少能活过童年。CLN1是 棕榈酰基蛋白硫酯酶编码基因PPT1功能缺失突变的结果 1(PPT1)。PPT1是一种溶酶体酶，负责从蛋白质中去除S棕榈酰基。如果是这样的话 功能失调，就像CLN1的情况一样，脂质和蛋白质在细胞中积累，并对 中枢神经系统、心脏和骨骼系统。目前还没有批准的治疗CLN1的方法，并且 患有这种疾病的儿童的平均预期寿命为8-12岁。目标：解决 CLN1疗法的关键未得到满足的需求，我们正在开发第一种小分子PPT1替代疗法 对这种疾病进行系统治疗。我们最近设计了一类新的脱胺丝裂原分子。 (DPALMS)，它化学选择性地从活细胞中的蛋白质中切割S-棕榈酰基。使用这项技术， 我们的目标是开发能够恢复CLN1细胞PPT1样硫酯酶活性的化合物。初步数据： 我们已经确定了一组DPALM先导化合物，它们能有效地使PPT1底物脱胺丝裂原并逆转 慢性非典型肺炎患者来源细胞中的蛋白质异常积聚。此外，我们还发现DPALMS是有效的 无毒剂量，在体内耐受性良好。具体目标：我们将优化药效和药效 我们的先导化合物的特性，以产生适合临床前试验和IND使能的DPALM 学习。在特定的目标1中，我们将使用联合合成和筛选生成一组优化的DPALM 努力。将使用模块化和强大的合成策略来生成新化合物，然后将 对PPT1底物的脱氨基转移活性进行筛选。在具体的AIM 2中，我们将评估疗效 在CLN1模型细胞系中优化的DPALM。将对化合物进行测试，以确定它们是否能够降低 在体外疾病模型中，积聚蛋白质，逆转溶酶体异常形态，抑制细胞凋亡。 拟议的研究将有助于开发高活性的DPALM，并加快 治疗慢性非典型肺炎的这种疗法的商业化。

PROJECT SUMMARY Significance: CLN1 is a devastating neurodegenerative disorder affecting 1 in 100,000 children born worldwide. This untreatable disease is characterized by dramatic developmental decline, neuromuscular symptoms, vision loss, and neuronal death. Unfortunately, patients with this disease rarely survive past childhood. CLN1 is the result of a loss-of-function mutation in the gene PPT1 that encodes for the enzyme palmitoyl-protein thioesterase 1 (PPT1). PPT1 is a lysosomal enzyme responsible for removing S-palmitoyl groups from proteins. When it is dysfunctional, as is the case with CLN1, lipids and proteins accumulate in cells and have toxic effects in the central nervous system, heart, and skeletal system. There are currently no approved therapies for CLN1, and children with this disease experience an average life expectancy of 8-12 years. Objective: To address the critical unmet need for CLN1 therapeutics, we are developing the first small molecule PPT1 replacement therapy for systemic treatment of this disease. We have recently engineered a novel class of depalmitoylating molecules (DPALMs), which chemoselectively cleave S-palmitoyl groups from proteins in live cells. Using this technology, we aim to develop compounds that can restore PPT1-like thioesterase activity to CLN1 cells. Preliminary Data: We have identified a set of DPALM lead compounds that effectively depalmitoylate PPT1 substrates and reverse abnormal protein accumulation in CLN1 patient-derived cells. Furthermore, we found that DPALMs are effective at non-toxic doses and are well tolerated in vivo. Specific Aims: We will optimize the potency and drug-like characteristics of our lead compounds to generate a DPALM suitable for preclinical testing and IND enabling studies. In SPECIFIC AIM 1, we will generate a set of optimized DPALMs using joint synthesis and screening efforts. A modular and robust synthetic strategy will be used to generate new compounds, which will then be screened for depalmitoylation activity toward PPT1 substrates. In SPECIFIC AIM 2, we will evaluate the efficacy of optimized DPALMs in CLN1 model cell lines. Compounds will be tested for their ability to reduce the level of accumulated proteins, reverse abnormal lysosome morphology, and inhibit apoptosis in in vitro disease models. The proposed studies will enable the development of a highly active DPALM and accelerate the commercialization of this therapy for the treatment of CLN1.