Nanodelivery of active NRTI to the central nervous system: humanized HIV murine m

活性 NRTI 纳米递送至中枢神经系统：人源化 HIV 小鼠 m

• 批准号: 7622398
• 负责人: SERGUEI V VINOGRADOV
• 金额: $ 20.09万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-29 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7622398
• 关键词: AIDS chemotherapy; AIDS neuropathy; Acquired Immunodeficiency Syndrome; Adverse effects; Amaze; Anti-Retroviral Agents; Antiviral Agents; Antiviral Therapy; Apolipoprotein E; Apoptotic; Biodistribution; Blood - brain barrier anatomy; Brain; Caliber; Cell model; Cells; Cessation of life; Chronic; Complication; Development; Didanosine; Dose; Drug Delivery Systems; Drug Formulations; Drug resistance; Encapsulated; Encephalitis; Environment; Goals; HIV; HIV-1; HIV-1 drug resistance; Highly Active Antiretroviral Therapy; In Vitro; Induction of Apoptosis; Infection; Inflammation; Injectable; Injection of therapeutic agent; Life; Measures; Mitochondria; Modality; Modeling; Mus; NanoGel; Neuraxis; Neurologic; Neurons; Neuropathy; Oral; Particle Size; Patients; Peptides; Peripheral; Permeability; Pharmaceutical Preparations; Public Health; Range; Reverse Transcriptase Inhibitors; System; Testing; Therapeutic; Toxic effect; Urticaria; Viral; Viral Load result; Virus; Zidovudine; chemotherapy; concept; drug efficacy; gastrointestinal epithelium; in vitro Model; in vivo; in vivo Model; innovation; macrophage; mortality; mouse model; mutant; nano; nanoformulation; neuron apoptosis; neuron loss; neuropathology; neurotoxic; neurotoxicity; novel; nucleoside analog; prospective; release factor; research study; response; tripolyphosphate; zidovudine triphosphate

DESCRIPTION (provided by applicant): Highly active anti-retroviral therapy (HAART) was capable to significantly reduce mortality and prolong lives of patients with AIDS. Nucleoside analogs (NA), aka reverse transcriptase inhibitors (NRTIs), are cornerstones of the HAART. Long-term treatment with high doses of NRTIs may result in various toxic side effects, and neurotoxicity is among the most severe complications. HIV-1 infection in the brain is accompanied by encephalitis (HIVE) that is poorly treated because of the impenetrability of the blood-brain barrier (BBB) for many drugs. Both neuropathic effects of HIV-1 and NRTIs are overlapping in the CNS. There is now an urgent need in new drugs or treatment modalities with ability to overcome the BBB and eradicate virus in the CNS while causing reduced neuronal toxicity. We introduce novel drug formulations with potential for systemic delivery of NRTIs to the brain, injectable Nano-NRTIs, which consist of 5'-triphosphates of NRTIs (NATP) packaged in polymeric nanogels. These Nano-NRTIs demonstrated antiviral effect even in drug-resistant HIV-1 mutants, lower neuronal apoptosis and mitochondrial toxicity than NRTIs. Injected naked or peptide-decorated nanogels could efficiently deliver NATP across the BBB and release the drug following their capture by macrophages in the brain. The central goal of this proposal is to evaluate antiviral efficacy and neurotoxicity- related parameters of two Nano-NRTIs in several cellular systems in vitro and in vivo. The hypothesis about therapeutic advantage of Nano-NRTIs will be tested in a humanized HIV-1 murine model and will allow us to determine quantitatively and qualitatively the efficacy of the CNS accumulation, inhibition of viral replication and neuronal damage, modeling the long-term NRTI drug treatment in one experiment. Our Specific Aim 1 is to compare reduction of viral load in cultured macrophages by antiviral NAs or nanogel-NATP formulations, using the neurotoxic NRTI, didanosine (ddI), and zidovudine (AZT), and their derivatives, ddI-TP and AZT-TP. 5'- Triphosphates will be synthesized from ddI and AZT, formulated in protective nanogel shells modified by the BBB-specific ApoE-peptide. Specific Aim 2 is to evaluate neurotoxic potential of antiviral NAs or nanogel-NATP formulations measuring mitochondrial toxicity and neuronal death in two cellular models, testing also alternative neurons-protective combinations of NATP with natural NTPs. Some nanogel-NTP formulations were found to increase survival of neurons in response to pro-apoptotic agents. Specific Aim 3: To validate humanized mouse model of HIV-1 infection in application to development of neuropathology (glial activation and neuronal loss) after anti-retroviral therapy and test the viral multiplication, mitochondrial toxicity and inflammation factor release resulting in neurotoxic complications. Positive "proof-of-the-concept" would become instrumental for further development of the Nano-NRTI approach to AIDS chemotherapy. PUBLIC HEALTH RELEVANCE: Highly active anti-retroviral therapy using nucleoside analogs (NRTIs), may result in severe neurotoxicity complications. HIV-1 infection in the brain is also accompanied by encephalitis that is poorly treated because of the impenetrability of the blood-brain barrier for many drugs. We introduce novel nanogel formulations with potential for systemic drug delivery to the brain, injectable Nano-NRTIs. Naked or brain-specific peptide- decorated nanogels can deliver activated drugs to infected macrophages in the brain. The central goal of this proposal is to evaluate and compare antiviral efficacy and neurotoxicity-related parameters of two Nano-NRTIs in several cellular systems in vitro and in a unique humanized HIV-1 murine model. We hypothesize that Nano- NRTIs will have therapeutic advantage over NRTIs and lower neurotoxicity. Positive "proof-of-the-concept" would open practical venues for further development of the Nano-NRTI approach to NeuroAIDS chemotherapy.

描述（由申请人提供）：高效抗逆转录病毒疗法（HAART）能够显著降低艾滋病患者的死亡率并延长其生命。核苷类似物（NA），又名逆转录酶抑制剂（NRTI），是HAART的基石。长期使用大剂量NRTIs可能导致各种毒副作用，其中神经毒性是最严重的并发症之一。脑中的HIV-1感染伴有脑炎（HIVE），由于许多药物无法穿透血脑屏障（BBB），因此治疗效果很差。HIV-1和NRTI的神经病变作用在CNS中重叠。现在迫切需要能够克服BBB并根除CNS中的病毒同时引起降低的神经元毒性的新药物或治疗方式。我们介绍了具有将NRTI全身递送至大脑的潜力的新型药物制剂，可注射的Nano-NRTI，其由包装在聚合物纳米凝胶中的NRTI的5 '-三磷酸盐（NATP）组成。这些Nano-NRTIs甚至在耐药HIV-1突变体中也表现出抗病毒作用，比NRTIs更低的神经细胞凋亡和线粒体毒性。注射裸或肽修饰的纳米凝胶可以有效地将NATP递送穿过BBB，并在被大脑中的巨噬细胞捕获后释放药物。本提案的中心目标是评价两种纳米NRTI在体外和体内几种细胞系统中的抗病毒功效和神经毒性相关参数。将在人源化HIV-1小鼠模型中测试关于Nano-NRTI治疗优势的假设，并使我们能够定量和定性确定CNS蓄积、病毒复制抑制和神经元损伤的疗效，在一项实验中模拟长期NRTI药物治疗。我们的具体目标1是比较使用神经毒性NRTI、去羟肌苷（ddI）和齐多夫定（AZT）及其衍生物ddI-TP和AZT-TP，通过抗病毒NAs或纳米凝胶-NATP制剂降低培养的巨噬细胞中的病毒载量。5 '-三磷酸盐将由ddI和AZT合成，配制在由BBB特异性ApoE肽修饰的保护性纳米凝胶壳中。具体目标2是评估抗病毒NA或纳米凝胶-NATP制剂的神经毒性潜力，测量两种细胞模型中的线粒体毒性和神经元死亡，还测试NATP与天然NTP的替代神经元保护组合。发现一些纳米凝胶-NTP制剂响应于促凋亡剂而增加神经元的存活。具体目标3：验证HIV-1感染的人源化小鼠模型在抗逆转录病毒治疗后神经病理学（胶质细胞活化和神经元丢失）发展中的应用，并检测病毒增殖、线粒体毒性和导致神经毒性并发症的炎症因子释放。积极的“概念验证”将有助于进一步开发艾滋病化疗的纳米NRTI方法。公共卫生相关性：使用核苷类似物（NRTI）的高效抗逆转录病毒治疗可能导致严重的神经毒性并发症。大脑中的HIV-1感染也伴随着脑炎，由于许多药物无法穿透血脑屏障，脑炎的治疗效果很差。我们介绍了新的纳米凝胶制剂，具有全身药物输送到大脑的潜力，可注射纳米NRTI。裸露的或脑特异性肽修饰的纳米凝胶可以将活化的药物递送到大脑中受感染的巨噬细胞。该提案的中心目标是评估和比较两种Nano-NRTI在体外几种细胞系统和独特的人源化HIV-1小鼠模型中的抗病毒疗效和神经毒性相关参数。我们推测纳米NRTIs将比NRTIs具有治疗优势和更低的神经毒性。积极的“概念验证”将为进一步开发NeuroAIDS化疗的Nano-NRTI方法开辟实际场所。