High throughput screen for discovery of N-acetyltransferase 8 Like (NAT8L) inhibitors

用于发现 N-乙酰转移酶 8 样 (NAT8L) 抑制剂的高通量筛选

基本信息

批准号：
10319002
负责人：
Barbara Stauch Slusher
金额：
$ 40.41万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-15 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10319002
关键词：
ABCB1 gene Acetyl Coenzyme A Acetyltransferase Address Anabolism Animals Antisense Oligonucleotides Arylalkylamine N-Acetyltransferase Aspartate Aspartoacylase Biological Assay Brain Brain Edema Canavan Disease Cells Cerebellum Cerebrum Cessation of life Chemicals Clinical Cognitive deficits Collection Data Demyelinations Deterioration Development Disease Dose Drug Kinetics Enzymes Exhibits Fluorescence Future Gene Deletion Genes Genetic Goals Human In Vitro Lead Libraries Macrocephaly Metabolic Molecular Molecular Bank Mus Mutation Myelin N-acetylaspartate Neuraxis Patients Penetration Permeability Pharmaceutical Chemistry Pharmaceutical Preparations Pre-Clinical Model Prion Diseases Property Prosencephalon Radioactivity Research Personnel Robotics Rodent Model Solubility Supportive care Symptoms Testing Therapeutic United States National Institutes of Health Vacuole analog aqueous assay development astrogliosis base carboxylate drug candidate drug discovery drug metabolism enzyme activity high throughput screening in vivo infancy inhibitor lead optimization leukodystrophy motor deficit mouse model myelination nervous system disorder neuropathology neurotransmission novel novel therapeutic intervention pharmacophore prevent process optimization repository response small molecule small molecule inhibitor small molecule libraries symptom management therapeutic candidate therapeutic development trend white matter

项目摘要

PROJECT SUMMARY Canavan disease (CD) is rare autosomal recessive leukodystrophy caused by mutations in the aspartoacylase gene (ASPA), leading to loss of enzyme activity and increased concentrations of the substrate N-acetylaspartate (NAA) in the brain. Accumulation of brain NAA results in spongiform degeneration of white matter, aberrant myelination, brain edema, macrocephaly, severe cognitive and motor deficits and ultimately death. There is no cure, nor is there a standard course of treatment for CD. Treatment is currently limited to supportive care and symptom management. Genetic deletion of the ASPA gene in mice has been shown to reproduce many of the CD disease symptoms seen in patients. Important to this proposal, deletion of the gene encoding for aspartate N-acetyltransferase 8 (NAT8L), the enzyme that catalyzes the biosynthesis of NAA from aspartate and acetyl CoA, prevented leukodystrophy in a CD mouse model. These mice showed substantial reduction in NAA levels and no evidence of astroglial vacuolation, astrogliosis, or demyelination in the cerebellum or forebrain. Similar therapeutic benefits were observed with intracisternal administration of antisense oligonucleotide to NAT8L. These results strongly suggest that inhibition of NAT8L would be useful in the treatment of CD. Currently, however, there are no NAT8L inhibitors that are clinically available; known NAT8L inhibitors have low potency (IC50 values in the μM - mM range) and/or possess carboxylate moieties that prevent brain penetration. This proposal aims to conduct high throughput screening (HTS) of a large and diverse 400,000 compound library for small molecule inhibitors of human NAT8L. Successful identification of tractable hit compounds followed by preliminary structural optimization should lead to the discovery of promising lead NAT8L inhibitors with potential for future development of therapeutics for CD. We are poised to seize this opportunity by executing the following three Specific Aims: (Aim 1) conduct NAT8L HTS and hit confirmation assays; (Aim 2) conduct hit clustering, preliminary SAR, and ADME profiling; (Aim 3) conduct preliminary lead optimization.

项目概要卡纳万病 (CD) 是一种罕见的常染色体隐性遗传性脑白质营养不良，由基因突变引起天冬氨酸酰基酶基因 (ASPA)，导致酶活性丧失和酶浓度增加底物 N-乙酰天冬氨酸 (NAA) 在大脑中积累，形成海绵状。白质变性、髓鞘形成异常、脑水肿、巨头畸形、严重认知和运动 CD 治疗没有治愈方法，也没有标准疗程。目前仅限于小鼠 ASPA 基因的支持性护理和症状管理。已被证明可以重现患者中出现的许多 CD 疾病症状，这对这一建议很重要，编码天冬氨酸 N-乙酰转移酶 8 (NAT8L) 的基因缺失，该酶催化从天冬氨酸和乙酰辅酶 A 生物合成 NAA 可以预防 CD 小鼠模型中的脑白质营养不良。小鼠的 NAA 水平显着降低，并且没有星形胶质细胞空泡化、星形胶质细胞增生或星形胶质细胞增生的证据小脑或前脑的脱髓鞘也观察到了脑池内的类似治疗效果。对NAT8L施用反义寡核苷酸这些结果强烈表明NAT8L的抑制。然而，目前尚无临床上的 NAT8L 抑制剂。可用；已知的 NAT8L 抑制剂效力较低（IC50 值在 μM - mM 范围内）和/或具有该提案旨在进行高通量筛选。 (HTS) 的一个大型且多样化的 400,000 个人类 NAT8L 小分子抑制剂化合物库。成功鉴定易处理的命中化合物，然后进行初步结构优化应该导致发现有前景的先导 NAT8L 抑制剂，具有未来治疗药物开发的潜力对于 CD，我们准备通过执行以下三个具体目标来抓住这个机会：（目标 1）进行 NAT8L HTS 和命中确认分析；（目标 2）进行命中聚类、初步 SAR 和 ADME 分析；（目标 3）进行初步先导优化。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

High Throughput Screening Cascade To Identify Human Aspartate N-Acetyltransferase (ANAT) Inhibitors for Canavan Disease.

高通量筛选级联鉴定卡纳万病的人天冬氨酸 N-乙酰转移酶 (ANAT) 抑制剂。

DOI：
10.1021/acschemneuro.1c00455
发表时间：
2021
期刊：
ACS chemical neuroscience
影响因子：
5
作者：
Nešuta,Ondřej;Thomas,AjitG;Alt,Jesse;Hin,Niyada;Neužilová,Anna;Long,Shunyou;Tsukamoto,Takashi;Rojas,Camilo;Wei,Huijun;Slusher,BarbaraS
通讯作者：
Slusher,BarbaraS