A Targeted Preemptive Approach to Addressing Mitochondrial Toxicity of Nucleoside

解决核苷线粒体毒性的有针对性的先发制人方法

基本信息

批准号：
8332894
负责人：
John K Buolamwini
金额：
$ 28.62万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2016-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8332894
关键词：
2&apos -fluoro-5-methylarabinosyluracil Address Adopted Adverse effects Antineoplastic Agents Antiviral Agents Area Attention Biochemical Biological Biological Assay Biology Biopharmaceutics Cardiomyopathies Cell surface Cells Chemicals Chromatography Clinical Cytoplasm Cytosol DNA biosynthesis DNA-Directed DNA Polymerase Data Deoxyguanosine kinase Developing Countries Development Didanosine Dipyridamole Disease Dose Drug Kinetics Drug usage Esters Fatty Liver Genetic HIV Hepatitis B Hepatitis B Virus Hepatotoxicity Highly Active Antiretroviral Therapy Infection Korea Lactic Acidosis Laser Scanning Confocal Microscopy Life Lipodystrophy Malignant neoplasm of pancreas Membrane Potentials Methodology Mitochondria Mitochondrial DNA Mitotic Molecular Multiple Sclerosis Myopathy Nucleoside Transporter Nucleosides Nucleotides Pancreatitis Parents Patients Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Pharmacotherapy Plague Polymerase Poverty Prevention strategy Prodrugs Property Protein Isoforms Purine Nucleosides Pyrimidine Nucleosides Rattus Research Reverse Transcriptase Inhibitors Solutions Stavudine Testing Therapeutic Tissues Toxic effect Transgenic Organisms United States Viral Cancer Virus Diseases Zidovudine acquired immunodeficiency analog anti-hepatitis B base drug development fialuridine gemcitabine human TK2 protein inhibitor/antagonist innovation interdisciplinary approach interest mitochondrial membrane novel novel strategies nucleoside analog nucleoside kinase overexpression preempt prevent research study success tandem mass spectrometry tool tripolyphosphate uptake

项目摘要

DESCRIPTION (provided by applicant): Although the introduction of highly active antiretroviral therapy (HAART) targeting the human immunodeficiency virus (HIV) revolutionized acquired immunodeficiency disease (AIDS) therapy, nucleoside reverse transcriptase inhibitors (NRTIs) that are a cornerstone of HAART cocktails have been found to cause severe mitochondrial toxicities that have limited their use. NRTIs cause toxicity by inhibiting mitochondrial DNA (mtDNA) synthesis through inhibition of polymerase g (Pol g), which is responsible for mtDNA synthesis. Although anti-HIV NRTIs have drawn the most attention to nucleoside drug-induced mitochondrial toxicity, this toxicity has also been seen with other antiviral nucleoside drugs such as fialuridine (FIAU) and clevudine (L-FMAU) that were being developed to treat hepatitis B (HBV) infections, and gemcitabine, the drug of choice for the treatment of pancreatic cancer. Moreover, the problem is likely to become more widespread as nucleoside drugs are used outside the antiviral and anticancer fields to disease areas such as multiple sclerosis. Unfortunately, there is no current proven general treatment of prevention strategy for nucleoside drug-induced mitochondrial toxicity. Thus, the development of innovative approaches to preempt or treat mitochondrial toxicity will be an important advance in the field of nucleoside drug therapy. NRTI triphosphates, the metabolites that inhibit Pol g can be synthesized inside mitochondria from imported NRTIs from the cytosol. Imported nucleosides are sequentially phosphorylated starting with rate-limiting monophosphorylation by mitochondrial-specific thymidine kinase 2 (TK-2) and/or deoxyguanosine kinase (dGK). Mitochondrial membrane nucleoside transporters (NTs) import NRTIs from the cytoplasm. The NT isoforms that have been implicated in mitochondrial nucleoside uptake are the equilibrative nucleoside transporters (ENTs), particularly ENT1 and ENT3. The transgenic overexpression of ENTs has been shown to enhance the mitochondrial toxicity of the former anti-hepatitis B nucleoside drug fialuridine (FIAU) and the NRTI AIDS drug AZT. We have obtained preliminary data showing that a prodrug approach could be used to protect against NRTI-induced mitochondrial toxicity; and we will test that hypothesis by pursuing the following specific aims: 1) synthesize and characterize novel prodrugs for lack of NT inhibition and cellular release of active drug, 2) evaluate the abiliy of prodrugs to protect mitochondria from the toxicity of nucleoside analog drugs, and 3) study biopharmaceutic and pharmacokinetic properties of selected prodrugs and their influence on the pharmacokinetics of nucleoside drugs of interest. A multidisciplinary approach integrating synthetic medicinal chemistry, biochemical and molecular biological assays, laser scanning confocal microscopy, chromatography and tandem mass spectrometry will be employed. The success of the project will confirm the possibility of protecting against nucleoside analog mitochondrial toxicity through the inhibition of nucleoside transporters, as an innovative general preemptive means of protection against toxicities of anti-HIV NRTIs and other nucleoside drugs like the anticancer nucleoside analog gemcitabine, which is associated with rare but potentially fatal liver toxicity and myopathy. Novel research tools will also be developed. PUBLIC HEALTH RELEVANCE: Highly active antiretroviral therapy (HAART) has been very useful for treating HIV/AIDS patients, but suffers from severe toxicities. The aim of this project s to develop a nucleoside transporter inhibition approach to protecting mitochondria, which get damaged by the nucleoside reverse transcriptase inhibitors present in HAART.

描述（由申请人提供）：虽然针对人类免疫缺陷病毒（HIV）的高效抗逆转录病毒疗法（HAART）的引入彻底改变了获得性免疫缺陷病（AIDS）疗法，但作为HAART鸡尾酒基石的核苷逆转录酶抑制剂（NRTI）已经已被发现会引起严重的线粒体毒性，从而限制了它们的使用。 NRTI 通过抑制负责 mtDNA 合成的聚合酶 g (Pol g) 来抑制线粒体 DNA (mtDNA) 合成，从而产生毒性。尽管抗 HIV NRTIs 最受关注的是核苷药物引起的线粒体毒性，但这种毒性也出现在其他抗病毒核苷药物中，例如例如正在开发用于治疗乙型肝炎 (HBV) 感染的菲阿尿苷 (FIAU) 和克拉夫定 (L-FMAU)，以及治疗胰腺癌的首选药物吉西他滨。此外，随着核苷类药物在抗病毒和抗癌领域之外用于多发性硬化症等疾病领域，这个问题可能会变得更加普遍。不幸的是，目前还没有经过验证的针对核苷药物引起的线粒体毒性的预防策略的一般治疗方法。因此，开发预防或治疗线粒体毒性的创新方法将是核苷药物治疗领域的重要进展。 NRTI 三磷酸盐是抑制 Pol g 的代谢物，可以通过从胞质溶胶导入的 NRTI 在线粒体内合成。导入的核苷通过线粒体特异性胸苷激酶 2 (TK-2) 和/或脱氧鸟苷激酶 (dGK) 从限速单磷酸化开始依次磷酸化。线粒体膜核苷转运蛋白 (NT) 从细胞质输入 NRTI。与线粒体核苷摄取有关的 NT 同工型是平衡核苷转运蛋白 (ENT)，特别是 ENT1 和 ENT3。 ENTs的转基因过度表达已被证明可以增强以前的抗乙型肝炎核苷药物菲阿尿苷（FIAU）和NRTI艾滋病药物AZT的线粒体毒性。我们获得的初步数据表明，前药方法可用于防止 NRTI 诱导的线粒体毒性；我们将通过追求以下具体目标来检验该假设：1）合成和表征新型前药，以消除 NT 抑制和活性药物的细胞释放，2）评估前药保护线粒体免受核苷类似物药物毒性的能力， 3) 研究所选前药的生物制药和药代动力学特性及其对感兴趣的核苷药物药代动力学的影响。将采用综合药物化学、生化和分子生物学测定、激光扫描共焦显微镜、色谱法和串联质谱法的多学科方法。该项目的成功将证实通过抑制核苷转运蛋白来防止核苷类似物线粒体毒性的可能性，作为一种创新的通用先发制人的方法来防止抗 HIV NRTI 和其他核苷药物（如抗癌核苷类似物吉西他滨）的毒性。与罕见但可能致命的肝毒性和肌病有关。还将开发新的研究工具。公共卫生相关性：高效抗逆转录病毒疗法 (HAART) 对于治疗 HIV/艾滋病患者非常有用，但存在严重的毒性。该项目的目的是开发一种核苷转运蛋白抑制方法来保护线粒体，避免受到 HAART 中核苷逆转录酶抑制剂的损害。