Ionic liquid-assisted drug delivery to brain reservoirs for treatment of neuroHIV

离子液体辅助药物输送至脑库治疗神经艾滋病毒

• 批准号: 10523303
• 负责人: Jason Richard Paris
• 金额: $ 43.55万
• 依托单位: UNIVERSITY OF MISSISSIPPI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10523303
• 关键词: ABCB1 gene; Acute; Affinity; Ames Assay; Anions; Anti-Retroviral Agents; Astrocytes; Attenuated; Biochemical; Biocompatible Materials; Biodistribution; Biological; Biological Assay; Blood; Blood - brain barrier anatomy; Brain; Brain region; CRISPR/Cas technology; Cations; Cell Adhesion; Cells; Central Nervous System; Central Nervous System Agents; Chemistry; Collaborations; Complete Blood Count; Complication; Data; Dependence; Dose; Drug Delivery Systems; Drug Efflux; Drug Kinetics; Endothelial Cells; Engineering; Erythrocytes; Formulation; Future; Genes; Gliosis; Goals; Gold; HIV; Heart; High Pressure Liquid Chromatography; Histopathology; Hour; Human; Immunohistochemistry; Impairment; In Vitro; Infection; Infusion procedures; Intracarotid; Intracarotid Infusion; Kidney; Kinetics; Lamivudine; Lead; Liquid substance; Liver; Lung; Macaca; Macaca mulatta; Measures; Mediating; Methods; Microglia; Modeling; Monitor; Monkeys; Morphine; Mutagenicity Tests; Neuroimmune; Neurons; Opiate Addiction; Opioid; Oral; Organ; Pathology; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacology; Plasma; Polymers; Precipitation; Primates; Prognosis; Proteins; Rattus; Reporter; Research; Rodent Model; Roentgen Rays; Route; SIV; Safety; Salts; Sampling; Spleen; Sprague-Dawley Rats; System; Techniques; Testing; Therapeutic; Time; Tissues; Toxic effect; Treatment Efficacy; Viral; Virus Diseases; Work; abacavir; antiretroviral therapy; astrogliosis; biomaterial compatibility; cell type; clinical development; cytokine; efficacy evaluation; improved; in vivo; in vivo Model; insight; light scattering; liquid chromatography mass spectrometry; liquid formulation; nano; nanoGold; nanoparticle; nanoparticle delivery; nanopolymer; near infrared dye; neuroAIDS; neuropathology; nonhuman primate; novel; opioid exposure; opioid use disorder; particle; preference; reproductive; safety assessment; small molecule; subcutaneous; success; uptake; vector; virology

PROJECT SUMMARY/ABSTRACT Despite the success of antiretroviral therapeutics (ARTs), they cannot eradicate HIV from reservoirs within the body, particularly those in the central nervous system (CNS). Moreover, opioids upregulate efflux transporters further promoting subtherapeutic concentrations of ART in the CNS. We have used biocompatible ionic liquids (ILs), molten salts comprised of asymmetric cations and anions, that can `tune' the affinity of nanoparticles to different cell types. Using this strategy, we have developed an IL with a balanced affinity for erythrocytes and microglia which promotes nanoparticle `hitchhiking' on erythrocytes to deliver them to the brain, and cells- selective targeting of microglia once delivered to the central compartment. Preliminary data in rats demonstrate ~48% of injected nanoparticles accumulating in the brain within 6 hours, a vast improvement over current nanoparticle delivery strategies. Preliminary analyses indicated that over 90% of CNS nanoparticles were associated with microglia. We have further demonstrated the capacity to load ART (abacavir) into nanoparticles which retained antiviremic efficacy when administered to HIV-infected human peripheral blood mononuclear cells (PBMCs). We hypothesize that we can further improve the tunable profile of our IL formulation to optimize cargo delivery to the brain and target additional cell types (including astrocytes). We anticipate that this cargo delivery strategy will be safe and efficacious in spite of CNS cell adhesion/transporter changes promoted by opioid exposure/dependence. To this end, we will (Aim 1) generate at least 5 novel ILs, in addition to our current lead, with varied cell-type affinity. We will confirm the preference that ILs confer to simian and human blood components as potential cargo carriers. (Aim 2) We will assess the safety (subacute, acute, subchronic, reproductive, mutagenic) and biodistribution of up to 5 novel ILs in rats that are opioid-naïve or opioid-dependent. ILs will be loaded with a premade scramble Cas9 vector with an eGFP reporter to confirm the capacity to deliver CRISPR-Cas9 constructs for potential HIV cure strategies. Safe ILs with a CNS-favorable biodistribution will be loaded with cART (abacavir, dolutegravir, and lamivudine) and assessed for efficacy in SIV-infected or HIV- infected simian or human PBMCs, respectively, or human microglia. All cells will be opioid-naïve or opioid- exposed. ILs with selectivity for microglia and astrocytes will be prioritized. (Aim 3) IL leads (based on CNS distribution and microglial/astrocytic selectivity) will be assessed for in vivo safety, biodistribution, and acute efficacy in a rhesus macaque model of SIV. In macaques, IL-assisted nanoparticles will be loaded with nanogold, in addition to cART, to confirm the time-course of biodistribution via X-ray. SIV infection will be monitored via CSF and blood draws. Complete blood count, chemistry, and cytokine profiling will be conducted on plasma and/or CSF. ART distribution will be confirmed via LC/MS. Gross histopathology will be conducted on organs and CNS tissues will be additionally assessed for microgliosis, astrogliosis, and sublethal neuronal damage. IL- assisted nanoparticles may realize the goal of achieving safe, cell-specific, CNS drug/cargo delivery.

项目概要/摘要 尽管抗逆转录病毒疗法（ART）取得了成功，但它们无法根除体内储存库中的艾滋病毒 身体，特别是中枢神经系统（CNS）。此外，阿片类药物上调外排转运蛋白 进一步促进中枢神经系统中 ART 的亚治疗浓度。我们使用了生物相容性离子液体 （IL），由不对称阳离子和阴离子组成的熔盐，可以“调节”纳米颗粒的亲和力 不同的细胞类型。利用这一策略，我们开发了一种对红细胞和红细胞具有平衡亲和力的 IL。 小胶质细胞促进纳米颗粒“搭便车”到红细胞上，将它们输送到大脑，而细胞- 一旦递送到中央区室，就选择性地靶向小胶质细胞。大鼠的初步数据表明 约 48% 的注射纳米粒子在 6 小时内积聚在大脑中，比目前的情况有了巨大的进步 纳米粒子递送策略。初步分析表明，90%以上的中枢神经系统纳米颗粒是 与小胶质细胞有关。我们进一步证明了将 ART（阿巴卡韦）负载到纳米颗粒中的能力 当给予感染艾滋病毒的人外周血单核细胞时，保留了抗病毒血症功效 （PBMC）。我们假设我们可以进一步改进 IL 配方的可调特性以优化货物 输送到大脑并靶向其他细胞类型（包括星形胶质细胞）。我们预计这批货物的交付 尽管阿片类药物促进中枢神经系统细胞粘附/转运蛋白发生变化，但该策略将是安全有效的 暴露/依赖。为此，除了我们目前的领先产品外，我们将（目标 1）生成至少 5 个新颖的 IL， 具有不同的细胞类型亲和力。我们将确认 IL 赋予猿猴和人类血液的偏好 组件作为潜在的货运载体。 （目标 2）我们将评估安全性（亚急性、急性、亚慢性、 多达 5 种新型 IL 在未使用阿片类药物或阿片类药物依赖的大鼠中进行生物分布。 IL 将装载带有 eGFP 报告基因的预制扰乱 Cas9 载体，以确认递送能力 CRISPR-Cas9 构建潜在的 HIV 治疗策略。具有有利于 CNS 的生物分布的安全 IL 装载 cART（阿巴卡韦、多替拉韦和拉米夫定）并评估其对 SIV 感染或 HIV-感染的疗效 分别感染猿猴或人类 PBMC，或人类小胶质细胞。所有细胞均未接触过阿片类药物或未接触过阿片类药物 裸露。对小胶质细胞和星形胶质细胞具有选择性的 IL 将被优先考虑。 （目标 3）IL 领先（基于 CNS 分布和小胶质细胞/星形细胞选择性）将评估体内安全性、生物分布和急性 在 SIV 恒河猴模型中的功效。在猕猴中，IL 辅助的纳米颗粒将装载纳米金， 除了 cART 之外，还通过 X 射线确认生物分布的时间过程。 SIV 感染将通过以下方式监测 脑脊液和抽血。将对血浆进行全血细胞计数、化学分析和细胞因子分析 和/或脑脊液。 ART 分布将通过 LC/MS 确认。将对器官进行大体组织病理学检查 中枢神经系统组织还将接受小胶质细胞增生、星形胶质细胞增生和亚致死神经元损伤的评估。白细胞介素- 辅助纳米颗粒可以实现安全、细胞特异性、中枢神经系统药物/货物输送的目标。