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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：
8628851
负责人：
John K Buolamwini
金额：
$ 12.31万
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 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8628851
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.

描述（由申请人提供）：尽管针对人类免疫缺陷病毒（HIV）的高效抗逆转录病毒治疗（HAART）的引入彻底改变了获得性免疫缺陷病（AIDS）治疗，但已发现作为HAART鸡尾酒基石的核苷逆转录酶抑制剂（NRTI）会导致严重的线粒体毒性，从而限制了其使用。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）合成而引起毒性。尽管抗HIV NRTI引起了对核苷类药物诱导的线粒体毒性的最大关注，但这种毒性也见于其他抗病毒核苷类药物，如正在开发用于治疗B肝炎（HBV）感染的氟替尼定（FIAU）和克来夫定（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以外的疾病领域（如多发性硬化症）的使用，这个问题可能会变得更加普遍。不幸的是，目前还没有被证实的核苷类药物诱导的线粒体毒性的预防策略的一般治疗。因此，开发创新方法来预防或治疗线粒体毒性将是核苷药物治疗领域的重要进展。抑制Pol g的代谢物NRTI三磷酸可以在线粒体内从来自细胞溶质的输入NRTI合成。输入的核苷从限速单磷酸化开始依次被肿瘤特异性胸苷激酶2（TK-2）和/或脱氧鸟苷激酶（dGK）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亚型是平衡型核苷转运蛋白（ENTs），特别是ENT 1和ENT 3。ENTs的转基因过表达已被证明可增强以前的抗B型肝炎核苷药物氟哌啶（FIAU）和NRTI AIDS药物AZT的线粒体毒性。我们已经获得了初步数据，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验证这一假设：1）合成和表征缺乏NT抑制和活性药物的细胞释放的新型前药，2）评价前药保护线粒体免受核苷类似物药物毒性的能力，和3）研究所选前药的生物药剂学和药代动力学性质及其对感兴趣的核苷药物的药代动力学的影响。将采用综合药物化学、生物化学和分子生物学测定、激光扫描共聚焦显微镜、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预防核苷类似物线粒体毒性的可能性，作为预防抗HIV NRTI和其他核苷药物（如抗癌核苷类似物吉西他滨）毒性的创新一般先发制人手段，这与罕见但可能致命的肝毒性和肌病有关。还将开发新型研究工具。