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 大脑中的底物 N-乙酰天冬氨酸 (NAA)。大脑中 NAA 的积累导致海绵状结构 白质变性、髓鞘形成异常、脑水肿、巨头畸形、严重认知和运动 赤字并最终导致死亡。 CD 无法治愈，也没有标准疗程。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 已被证明可以重现患者中出现的许多 CD 疾病症状。对于这个提案来说很重要的是， 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。这些 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 小脑或前脑脱髓鞘。脑池内治疗也观察到类似的治疗效果 将反义寡核苷酸施用至NAT8L。这些结果强烈表明 NAT8L 的抑制 将有助于 CD 的治疗。但目前尚无 NAT8L 抑制剂进入临床。 可用的;已知的 NAT8L 抑制剂效力较低（IC50 值在 μM - mM 范围内）和/或具有 阻止大脑渗透的羧酸部分。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 和命中确认测定； （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

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