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.

项目摘要/摘要 尽管抗逆转录病毒疗法（ART）取得了成功，但它们仍无法从储层中脱离艾滋病毒 身体，特别是中枢神经系统（CNS）的身体。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艺术的亚治疗浓度。我们使用了生物相容性的离子液体 （ils），由不对称阳离子和阴离子组成的熔融盐，可以“调整”纳米颗粒与 不同的细胞类型。使用此策略，我们开发了一个IL，对红细胞和红细胞具有平衡的亲和力 小胶质细胞促进红细胞上纳米颗粒的“搭便车”，以将其输送到大脑，并将细胞传递到大脑中 - 小胶质细胞的选择性靶向一旦传递到中央车厢。大鼠的初步数据证明 〜48％的注射纳米颗粒在6小时内积聚在大脑中，比电流大大改善 纳米颗粒输送策略。初步分析表明，超过90％的CNS纳米颗粒为 与小胶质细胞相关。我们进一步证明了将艺术品（Abacavir）加载到纳米颗粒的能力 当对HIV感染的人外周血单核细胞施用时，它保留了抗病毒效率 （PBMC）。我们假设我们可以进一步改善IL公式的可调曲线以优化货物 传递到大脑并靶向其他细胞类型（包括星形胶质细胞）。我们预计这家货物交付 尽管CNS细胞粘合剂/转运蛋白的变化是由Oopioid促进的，策略将是安全有效的 暴露/依赖。为此，我们（AIM 1）除了我们当前的潜在客户外，还将至少生成5个新颖的IL， 具有不同的细胞类型亲和力。我们将确认ILS赋予猿猴和人类血液的偏爱 组件作为潜在的货物载体。 （AIM 2）我们将评估安全性（子曲线，急性，亚基， 在不依赖阿片类药物或阿片类药物的大鼠中，多达5个新颖的IL的生殖，诱变）和生物分布。 ILS将装有带有EGFP记者的预制争夺Cas9矢量，以确认交付的能力 CRISPR-CAS9构造，用于潜在的HIV治疗策略。具有CNS有利生物分布的安全性IL将是 装有购物车（Abacavir，Dolutegravir和Lamivudine），并评估了SIV感染或HIV的效率 分别感染的邻苯二豆或人PBMC或人类小胶质细胞。所有细胞都将是卵毒药 - 不接受或卵虫类药物 裸露。对小胶质细胞和星形胶质细胞的选择性的IL将优先考虑。 （AIM 3）IL引线（基于CNS 将评估分布和小胶质细胞选择性）的体内安全性，生物分布和急性 SIV的恒河猕猴模型中的功效。在猕猴中，IL辅助纳米颗粒将装有纳米质量， 除了购物车，还可以通过X射线确认生物分布的时间顺序。 SIV感染将通过 CSF和血液吸收。将在等离子体上进行全血数，化学和细胞因子分析 和/或CSF。艺术分布将通过LC/MS确认。总体组织病理学将在器官上进行 和中枢神经系统组织将进一步评估小胶质细胞增多症，星形胶质细胞增多症和神经元损伤。 il- 辅助纳米颗粒可能意识到实现安全，细胞特异性的CNS药物/货物输送的目标。