Mannosidase inhibitors as therapeutics for glycoprotein misfolding diseases

甘露糖苷酶抑制剂作为糖蛋白错误折叠疾病的治疗药物

基本信息

批准号：
7665168
负责人：
KELLEY W. MOREMEN
金额：
$ 48.61万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-15 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7665168
关键词：
Address Adverse effects Affinity Alpha-mannosidase Binding Biochemical Biochemistry Biological Assay Biological Models Carbohydrates Cell Maturation Cell surface Cells Chemicals Combinatorial Synthesis Complex Coupled Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Cytoplasm Defect Destinations Disease Disease model Effectiveness Endoplasmic Reticulum Enzyme Inhibition Enzymes Estrogen Receptor alpha Evaluation Glycoproteins Glycoside Hydrolases Goals Golgi Apparatus Hereditary Disease Human Human Genetics In Vitro Kinetics Lead Lectin Libraries Lysosomal Storage Diseases Lysosomes Mannose Mannosidase Medicine Modeling Modification Molecular Chaperones Mutation NIH Program Announcements Oligosaccharides Pathology Play Polysaccharides Process Protein C Inhibitor Proteins Quality Control Recombinants Research Research Personnel Role Screening procedure Signal Transduction Specificity Structure Study models Testing Therapeutic Time Universities analog base college combinatorial cytotoxicity enzyme activity high throughput screening human disease inhibitor/antagonist loss of function mutant novel therapeutics overexpression polypeptide premature programs protein folding protein misfolding residence response small molecule small molecule libraries structural biology virtual

项目摘要

DESCRIPTION (provided by applicant): The long-term goals of this application are to develop selective inhibitors for an ER glycoprotein processing enzyme that plays a key role in quality control in the endoplasmic reticulum (ER) as broad-based therapeutics for glycoprotein misfolding diseases. This enzyme, ERalpha-mannosidase I (ERManl), acts as a key timer for ER residence for newly synthesized glycoproteins by initiating a rate-limiting step leading to a cascade of interactions that ultimately leads to the targeting of terminally misfolded glycoproteins for retrotranslocation to the cytoplasm and proteasomal disposal in a process known as "ER-associated degradation" (ERAD). Many loss-of-function human genetic diseases result from mutations that cause delayed protein folding kinetics rather than generating terminally misfolded polypeptides. Recognition of the incompletely folded intermediates by the ERAD targeting machinery can lead to premature disposal of potentially functional glycoproteins and subsequently leads to pathology. Inhibition of the rate-determining steps in ERAD could provide a broad-based therapeutic approach for treatment of glycprotein misfolding diseases by delaying ERAD and providing sufficient time to complete the protein folding process. All of the known inhibitors of early mannose trimming steps, however, also have unacceptable serious side effects. They also inhibit glycan processing alpha-mannosidases in the Golgi complex and block maturation to complex type glycan structures on cell surface and secreted glycoproteins. Thus, the goals of this application are to identify selective ERManl inhibitors that can act to delay ERAD, rescue ER protein folding defects in human disease, and retain normal glycan maturation in the Golgi complex. The unique interdisciplinary team that we have assembled takes advantage of ongoing synergistic collaborative interactions between investigators at the University of Georgia and Baylor College of Medicine with expertise in the synthesis of selective glycosidase inhibitors (Boons), the biochemistry and structural biology of the ER and Golgi mannosidases (Moremen), and cell-based assays for a human glycoprotein misfolding disorder, alpha1-antitrypsin deficiency (Sifers). Promising leads will also be evaluated in established lysosomal storage disease models by collaborators (Amicus). Three specific aims are proposed including 1) the directed rational and combinatorial synthesis of analogs of alpha-mannosidase inhibitors with selectivity toward ERManl, 2) high-throughput screens combined with detailed biochemical and structural analysis to assess selectivity and effectiveness of the inhibitor compounds in blocking ERManl but not Golgi glycan maturation, and 3) cell-based assays to assess chemical chaperone effects of mannosidase inhibitors in the rescue of mutant alpha1-antitrypsin secretion and lysosomal enzyme targeting and without blockage of N-glycan maturation.

描述（由申请人提供）：本申请的长期目标是为ER糖蛋白加工酶开发选择性抑制剂，该酶在内质网（ER）中起着关键作用，作为基于糖蛋白释放蛋白疾病的广泛疗法。这种酶，Eralpha-甘露糖苷I（Ermanl），通过启动限制速率的步骤，是新合成的糖蛋白的关键计时器，导致一系列相互作用，导致相互作用最终导致针对终止脱氧化糖蛋白的靶向量身定性的囊肿，以使其对远程驱动器的反构素化的量表进行了反应，并导致均具有较大的糖蛋白，并将其置入量的均可分配量。 “与ER相关的降解”（ERAD）。许多功能丧失的人遗传疾病是由于突变导致延迟蛋白质折叠动力学而不是产生终末错误折叠多肽引起的。识别通过射击靶向机械对不完全折叠的中间体的识别会导致可能过早处理潜在的功能性糖蛋白，并随后导致病理学。通过延迟ERAD并提供足够的时间来完成蛋白质折叠过程，可以抑制ERAD中的速率确定步骤可以提供一种基于广泛的治疗方法来治疗糖蛋白错误折叠疾病。但是，所有已知的早期甘露糖修剪步骤的抑制剂也都具有不可接受的严重副作用。它们还抑制高尔基体复合物中的聚糖加工α-甘露糖苷酶，并阻止细胞表面和分泌糖蛋白上的复杂类型聚糖结构的成熟。因此，本应用的目标是确定可以延迟ERAD，挽救ER蛋白质折叠缺陷的选择性ERMANL抑制剂，并保留高尔基体综合体中正常的聚糖成熟。我们已经组装的独特跨学科团队利用了佐治亚大学研究人员与贝勒医学学院的研究人员之间的持续协同合作互动，并具有选择性糖苷酶抑制剂（BOONS）的专业知识错误折叠障碍，Alpha1-抗抗抑制蛋白缺乏症（Sifers）。合作者（Amicus）在既定的溶酶体存储疾病模型中也将评估有希望的潜在客户。提出了三个具体目的，包括1）对α-甘露糖苷酶抑制剂的类似物的定向合理和组合合成，对Ermanl的选择性具有选择性，2）高通量筛选，结合详细的生物化学和结构分析，以评估抑制剂的选择性和抑制剂化合物的选择性和有效性。甘露糖苷酶抑制剂在拯救突变α1-抗抗蛋白酶分泌和溶酶体酶靶向且无n-聚糖成熟的情况下的伴侣作用。