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：
8955457
负责人：
John K Buolamwini
金额：
$ 17.14万
依托单位：
ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
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 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8955457
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）的高度活性抗逆转录病毒疗法（HAART）彻底改变了可获得的免疫缺陷疾病（AIDS）治疗，核苷逆转录酶抑制剂（NRTI）（NRTIS）引起了Haart cocktients cocktien cockities的强度定毒性，使其引起了Haart cockities的限制性。 NRTI通过抑制线粒体DNA（mtDNA）合成而通过抑制聚合酶G（Pol G）引起毒性，该聚合酶G（pol G）负责MTDNA合成。尽管抗HIV NRTI最吸引了对核苷药物诱导的线粒体毒性的关注，但这种毒性也与其他抗病毒核苷药物一起观察到了这种毒性作为用于治疗丙型肝炎（HBV）感染的Fialuridine（Fiau）和Clevudine（L-FMAU），以及吉西他滨（Gemcitabine），这是治疗胰腺癌的首选药物。此外，由于核苷药物在抗病毒药和抗癌领域以外用于疾病区域（例如多发性硬化症），因此问题可能会变得更加普遍。不幸的是，目前尚无对核苷药物诱导的线粒体毒性的预防策略的一般治疗。因此，在核苷药物治疗领域，抢先或治疗线粒体毒性的创新方法的发展将是重要的进步。 NRTI三磷酸盐，抑制pol G的代谢产物可以在线粒体内与从细胞质进口的NRTIS合成。从线粒体特异性胸苷激酶2（TK-2）和/或脱氧鸟苷激酶（DGK）开始，进口的核苷是依次磷酸化的磷酸化。线粒体膜核苷转运蛋白（NTS）从细胞质中导入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（尤其是ENT1和ENT3）。已显示ETENT的转基因过表达可增强前抗肝炎B核苷药物Fialuridine（FIAU）和NRTI AIDS药物AZT的线粒体毒性。我们获得了初步数据，表明可以使用前药方法来防止NRTI诱导的线粒体毒性。我们将通过追求以下特定目的来检验该假设：1）合成和表征缺乏NT抑制作用和活性药物的细胞释放的新型前药，2）评估前药的abiliy在保护线粒体以保护线粒体以免受核苷的毒性的影响，并研究了核素类模拟药物的影响和3）研究了药物的影响，并将其选择性化属性构成药物剂的质量和药物的影响。感兴趣的核苷药物。多学科方法将采用合成药物化学，生化和分子生物学测定，激光扫描共聚焦显微镜，色谱和串联质谱法。该项目的成功将证实有可能通过抑制核苷转运蛋白来预防核苷类似线粒体毒性，这是一种创新的一般预先预先预先预先保护抗HIV NRTIS和其他核苷药物（例如抗癌药物核苷类似物的脂肪及其脂肪效率）的抗HIV NRTI和其他核苷药物的毒性。新颖的研究工具也将开发。