Ultrasound-triggered drug delivery with acoustically active red blood cells

利用声学活性红细胞进行超声触发药物输送

• 批准号: 8637280
• 负责人: ALEXANDER L KLIBANOV
• 金额: $ 23.33万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8637280
• 关键词: Acoustics; Adhesions; Antibiotics; Antibodies; Apolipoprotein E; Area; Binding; Biodistribution; Blood; Blood Circulation; Blood Vessels; Bolus Infusion; C57BL/6 Mouse; Cardiovascular Diseases; Cause of Death; Cell Culture Techniques; Cell Proliferation; Cholesterol; Clinical; Clinical Trials; Contrast Media; Detection; Diagnosis; Diet; Disease; Doxorubicin; Doxycycline; Drug Carriers; Drug Delivery Systems; Drug Formulations; Dyes; Equipment; Erythrocytes; Exhibits; Fluorescence; Fluorescence Microscopy; Fluorocarbons; Focused Ultrasound Therapy; Frequencies; Gases; Generations; Half-Life; High Prevalence; Image; In Vitro; Infusion procedures; Injection of therapeutic agent; Label; Laboratories; Ligands; Lipids; Liquid substance; Longevity; Microbubbles; Modeling; Molecular Target; Monitor; Mus; Patients; Perfusion; Pharmaceutical Preparations; Physiologic pulse; Pilot Projects; Production; Property; Route; Speed; Surface; Therapeutic; Therapeutic Index; Time; Tissues; Treatment Efficacy; Ultrasonography; Vascular Cell Adhesion Molecule-1; allergic response; antiproliferative drugs; base; biomaterial compatibility; clinically relevant; fluorexon; improved; in vivo; inflammatory marker; molecular imaging; monolayer; mortality; nanoDroplet; novel; particle; pressure; public health relevance; research study; response; targeted delivery; tool

DESCRIPTION (provided by applicant): Vascular imaging and therapy are important tools in management of cardiovascular diseases. Ultrasound can image and non-invasively deliver therapy. Microbubbles are intravascular ultrasound contrast agents that provide blood pool imaging in real time and targeted (molecular) imaging or drug delivery vehicles with ultrasound activation. Microbubbles have a short half-life; therapeutic payload is small. Red blood cells have been loaded with drugs and reinjected in the bloodstream, including successful clinical trials. They circulate for weeks and have high therapeutic payload, but have poor acoustic contrast and lack a mechanism to trigger drug release. We propose a new ultrasound triggered drug delivery vehicle - acoustically active RBCs (AARBCs) - combining longevity, drug payload and targetability of RBCs with acoustic activation. AARBC is built as RBC with gas precursor perfluorocarbon (PFC) liquid nanodroplet inside. Entrapped PFC absorbs acoustic energy, converts to gas, which triggers drug release from RBC. Particle surface is natural; an early-generation AARBC shows ultrasound contrast half-life in mice ~15 min, longer than microbubbles. Specific Aims. Aim #1. Demonstrate and optimize acoustically active red blood cells as vascular contrast agents with extended circulation lifetime. Production of AARBCs will be optimized to maximize PFC entrapment and acoustic triggering. The acoustic response of AARBCs will be assessed at clinically relevant ultrasound conditions. AARBCs will be administered to mice to characterize contrast longevity, biodistribution and biocompatibility. Aim #2. Develop triggered drug delivery vehicles from AARBC. AARBCs will be loaded with calcein, doxorubicin or doxycycline. Contents release and drug delivery will be investigated after insonation, by fluorescence microscopy and via supernatant fluorescence. Therapeutic efficacy will be investigated in a cell culture drug delivery model and in murine vasculature. Aim #3. Achieve molecular targeting of drug-loaded AARBC. Antibody against VCAM-1 will be attached to AARBCs by noncovalent anchoring and PEG tether. Flow chamber experiments will verify binding of AARBCs to VCAM-1. AARBC targeting to VCAM-1 will be assessed by contrast ultrasound imaging in ApoE -/- mice on high-cholesterol diet, followed by ultrasound-triggered image-guided AARBC release of contents, and a fluorescent drug (doxycycline) detection in the vessel wall.

描述（由申请人提供）：血管成像和治疗是心血管疾病管理的重要工具。超声可以形象和非侵入性提供治疗。微泡是血管内超声对比剂，可实时提供血液池成像，并具有超声激活的靶向（分子）成像或药物输送车辆。微泡的半衰期很短；治疗有效载荷很小。红细胞已经加载了药物并重新注射在血液中，包括成功的临床试验。他们循环数周，具有高治疗有效载荷，但声学对比度较差，缺乏触发药物释放的机制。我们提出了一种新的超声触发药物输送车 - 声学活跃的RBC（AARBC） - 将RBC的寿命，药物有效载荷和原声激活结合起来。 AARBC用RBC建造，内部具有气体前体的全氟碳（PFC）液体纳米光。被夹住的PFC吸收声学能量，转化为气体，从而触发了RBC的药物释放。粒子表面是自然的；早期的AARBC在约15分钟的小鼠中显示出超声对比半衰期，比微泡长。具体目标。目标＃1。证明并优化了具有延长循环寿命的血管对比剂的声学活性红细胞。 AARBC的生产将被优化，以最大程度地提高PFC捕集和声学触发。 AARBC的声音响应将在临床相关的超声条件下进行评估。 AARBC将用于小鼠，以表征对比度寿命，生物分布和生物相容性。目标＃2。从AARBC开发触发的药物。 AARBC将加载钙牛，阿霉素或强力霉素。含量释放和药物递送将在事故后通过荧光显微镜和上清液荧光研究。治疗功效将在细胞培养药物递送模型和鼠血管系统中进行研究。目标＃3。实现载有药物的AARBC的分子靶向。针对VCAM-1的抗体将通过非共价锚定和钉子系连接到AARBC。流室实验将验证AARBC与VCAM-1的结合。 AARBC针对VCAM-1的AARBC将通过高胆固醇饮食中的ApoE - / - 小鼠的对比度超声成像进行评估，然后进行超声触发的图像引导的AARBC含量的释放，并在容器壁中检测荧光药物（多克环蛋白）检测。