Studies on Probenecid Prodrugs that Protect against Mitochondrial Toxicity of Tenofovir

预防替诺福韦线粒体毒性的丙磺舒前药研究

• 批准号: 10672238
• 负责人: John K Buolamwini
• 金额: $ 19.5万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10672238
• 关键词: Acute; Address; Adopted; Alternative Therapies; American; Anti-HIV Agents; Biochemistry; Biological; Blood; Cardiac; Case Study; Cell membrane; Cells; Clinical; Development; Disease; Drug usage; Epithelial Cells; Etiology; Event; Excretory function; Exhibits; Fanconi Syndrome; Fumarates; Functional disorder; Genetic; Goals; HIV; HIV Infections; Hepatitis B Virus; Highly Active Antiretroviral Therapy; Human; Individual; Kidney; Kidney Diseases; Lead; Lipids; Marketing; Measurement; Metabolic syndrome; Methods; Mitochondria; Mitochondrial DNA; Mitochondrial Matrix; Molecular Biology; Mother-to-child HIV transmission; Nucleosides; Nucleotides; Organic Anion Transporters; Oxidative Stress; Parents; Pathway interactions; Patients; Persons; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Prevention; Probenecid; Process; Prodrugs; Production; Property; Proximal Kidney Tubules; Reactive Oxygen Species; Regimen; Renal function; Research; Rest; Reverse Transcriptase Inhibitors; Risk; Route; Services; Specialist; Structure-Activity Relationship; Tenofovir; Testing; Therapeutic; Time; Toxic effect; Tubular formation; Vaccines; Vertebral column; Virus Diseases; Weight Gain; Western Blotting; acquired immunodeficiency; basolateral membrane; diabetes risk; fasting glucose; functional genomics; in vivo; inhibitor; innovation; insight; interdisciplinary approach; kidney dysfunction; lead optimization; mitochondrial dysfunction; mitochondrial membrane; nephrotoxicity; novel; off-patent; pharmacologic; pre-exposure prophylaxis; prevent; prophylactic; stem; success; tool; uptake

ABSTRACT: Almost 38 million individuals worldwide are infected with the human immunodeficiency virus (HIV) causative agent of acquired immunodeficiency disease (AIDS), 1.2 million of them being Americans. With no vaccine and no cure available, HIV/AIDS patients must take anti-HIV drugs throughout the rest of their lives. Thus, the long-term use of highly active antiretroviral therapy (HAART)drugs that revolutionized HIV/AIDS AIDS patients can also live as long as non-infected people, has been accompanied by serious mitochondrial toxicities, with fatal consequences that led to discontinuation of many of the early effective nucleoside reverse transcriptase (RT) inhibitor (NRTIs) backbone components, like ddC and d4T. The development of the nucleotide RT inhibitors (NtRTIs) particularly tenofovir (TFV), has been innovative in the HIV RT inhibitors field, to the extent that it has gained widespread use worldwide and is incorporated into almost all first-line HIV/AIDS therapeutic combination regimens, as well as used for pre-exposure prophylactics (PrEP). TFV is marketed as two prodrugs tenofovir disoproxil fumarate (TDF) and tenofovir alafenamide (TAF). However, long-term use of the TDF prodrug causes kidney toxicity stemming from the active parent TFV. TFV-associated kidney toxicity is said to have constituted the single referrals for specialist renal services when TFD was first introduced. Moreover, this renal decline was irreversible in about 30 % of patients even after cessation of TFV therapy. Although, TAF, which was introduced more recently is more potent and less toxic than TDF, case studies show it has kidney toxicity potential which may manifest upon long-term use. Moreover, it causes troubling weight gain that could exacerbate metabolic syndrome. These problems mean that TAF cannot always replace TDF. The mechanisms of TFV toxicity are not well understood, but damage to renal proximal tubular mitochondrial has been strongly implicated. The goal of this research is to gain better understanding and develop effective approaches to addressing TFV kidney toxicity. The specific aims are: 1) to confirm that human mitochondrial membranes express functional organic anion transporters 1 and 3 (hOAT1 and hOAT3), and show their ability to import TFV into mitochondria, and, 2) to develop and investigate the protective ability of mitochondrially targeted hOAT1 and 3 inhibitor prodrugs against TFV toxicity. We have demonstrated the expression of the TFV transporters hOAT1 and hOAT3 in the mitochondrial membranes of human renal proximal tubule epithelial cells, supporting our hypothesis that TFV enters mitochondria through hOAT1 and 3 to cause mitochondrial toxicity. For aim 2, we have synthesized an initial hOAT1, and 3 inhibitor prodrug and shown that it is protective against TFV mitochondrial toxicity. A comprehensive multidisciplinary approach involving medicinal chemistry, biochemistry, functional genomics, and pharmacology will be applied, using human kidney proximal tubule cells. Our initially synthesized targeted prodrug reverses TFV-induced mitochondrial toxicity. The success of this project will give new insights into TFV-induced toxicity and provide biological probes and/or potential drug leads.

摘要：全球近 3800 万人感染人类免疫缺陷病毒 (HIV) 获得性免疫缺陷病（艾滋病）的病原体，其中 120 万是美国人。没有 由于没有疫苗和治愈方法，艾滋病毒/艾滋病患者必须终生服用抗艾滋病毒药物。因此， 长期使用高效抗逆转录病毒治疗 (HAART) 药物彻底改变了艾滋病毒/艾滋病 艾滋病患者也可以和未感染者一样长寿 伴随严重的线粒体毒性，造成致命后果，导致许多药物停产 早期有效的核苷逆转录酶 (RT) 抑制剂 (NRTI) 主干成分，如 ddC 和 d4T。这 核苷酸 RT 抑制剂 (NtRTIs) 特别是替诺福韦 (TFV) 的开发在 HIV RT 领域具有创新性 抑制剂领域，已在全球范围内得到广泛应用，并被纳入几乎所有一线药物 HIV/AIDS 治疗组合方案，以及用于暴露前预防 (PrEP)。 TFV 是 作为两种前药富马酸替诺福韦二吡呋酯 (TDF) 和替诺福韦艾拉酚胺 (TAF) 上市。然而，长期来看 使用 TDF 前药会导致由活性母体 TFV 引起的肾毒性。 TFV 相关肾毒性 据说构成了单一 转介专科肾脏服务 TFD 首次引入时。此外，即使在治疗后，约 30% 的患者的肾功能衰退也是不可逆转的。 停止 TFV 治疗。尽管最近推出的 TAF 比 TDF，案例研究表明它具有潜在的肾脏毒性，长期使用可能会显现出来。而且，它会导致 令人不安的体重增加可能会加剧代谢综合征。这些问题意味着 TAF 不能总是 更换 TDF。 TFV 毒性机制尚不清楚，但对肾近端肾小管的损害 线粒体密切相关。本研究的目的是为了更好地理解和发展 解决 TFV 肾毒性的有效方法。具体目标是：1）确认人类 线粒体膜表达功能性有机阴离子转运蛋白 1 和 3（hOAT1 和 hOAT3），并显示 他们将 TFV 输入线粒体的能力，以及 2) 开发和研究 TFV 的保护能力 线粒体靶向 hOAT1 和 3 抑制剂前药可对抗 TFV 毒性。我们已经证明了 人肾近端线粒体膜中 TFV 转运蛋白 hOAT1 和 hOAT3 的表达 肾小管上皮细胞，支持我们的假设，即 TFV 通过 hOAT1 和 3 进入线粒体，导致 线粒体毒性。对于目标 2，我们合成了初始 hOAT1 和 3 抑制剂前药，并证明它是 防止 TFV 线粒体毒性。涉及医学的综合性多学科方法 将应用化学、生物化学、功能基因组学和药理学，利用人类肾近端 肾小管细胞。我们最初合成的靶向前药可逆转 TFV 诱导的线粒体毒性。的成功 该项目将为 TFV 诱导的毒性提供新的见解，并提供生物探针和/或潜在的药物先导化合物。