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

• 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
• 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

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.

项目摘要 Canavan病（CD）是由突变引起的罕见常染色体隐性白细胞营养不良。 asparto酰基酶基因（ASPA），导致酶活性的损失和增加的浓度 大脑中的底物N-乙酰天冬氨酸（NAA）。大脑NAA的积累导致赞助 白质，异常髓鞘，脑水肿，脑畸形，严重认知和运动的变性 缺陷，最终死亡。没有治愈方法，也没有标准的CD治疗方法。治疗 目前仅限于支持性护理和症状管理。小鼠ASPA基因的遗传缺失 已证明可以重现患者看到的许多CD疾病症状。对此提议很重要， 对于天冬氨酸N-乙酰基转移酶8（NAT8L）编码的基因删除，该酶催化了催化的酶 天冬氨酸和乙酰基COA的NAA生物合成可防止CD小鼠模型中的白细胞营养不良。这些 小鼠显示NAA水平的大幅降低，没有星形胶质真空化，星形胶质细胞增生或 小脑或前脑中的脱髓鞘。与肠内观察到类似的治疗益处 对NAT8L的反义寡核苷酸给药。这些结果强烈表明抑制NAT8L 对于CD的治疗将是有用的。但是，目前尚无临床上的NAT8L抑制剂 可用的;已知的NAT8L抑制剂的效力较低（μm -mm范围内的IC50值）和/或拥有 羧酸盐部分可防止大脑渗透。该建议旨在进行高吞吐量筛查 人Nat8l的小分子抑制剂的大型和多样化的40万种化合物库的（HTS）。 成功鉴定可易访问的命中化合物，然后进行初步结构优化 导致发现有希望的铅NAT8L抑制剂，并有可能未来的治疗发展 对于CD。通过执行以下三个特定目标，我们被毒死了以抓住这一机会：（目标1） 进行NAT8L HTS并命中确认测定； （AIM 2）进行命中率，初步SAR和ADME 分析； （目标3）进行初步铅优化。

项目成果

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