Celecoxib Derivative: Host Cell-Directed Inhibitors of Intracellular Pathogens

塞来昔布衍生物：宿主细胞定向的细胞内病原体抑制剂

• 批准号: 8391486
• 负责人: Kristy M Ainslie
• 金额: $ 18.78万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-17 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8391486
• 关键词: Acids; Aerosols; Affect; Aftercare; Animal Model; Animals; Antibiotics; Autophagocytosis; Bacteria; Biocompatible Materials; Biopolymers; Blood; Breathing; Cell Survival; Cells; Cessation of life; Collection; Dextrans; Dose; Drug Delivery Systems; Drug resistance; Encapsulated; Environment; Evaluation; Fluorescence Spectroscopy; Francisella tularensis; Frequencies; Grant; Granuloma; Hematology; Histopathology; Human; Hydrophobicity; Image; Immune; In Vitro; Infection; Injection of therapeutic agent; Intervention; Kinetics; Legionella pneumophila; Liver; Lymphocyte; Lymphoid Tissue; Measurement; Minimum Inhibitory Concentration measurement; Multi-Drug Resistance; Multidrug-Resistant Tuberculosis; Mus; Mycobacterium tuberculosis; National Institute of Allergy and Infectious Disease; Nature; Needles; Nose; Organ; Organ Survival; Outcome; PTGS2 gene; Particle Size; Particulate; Pathology; Pathway interactions; Penetration; Phagocytes; Phagosomes; Pharmaceutical Preparations; Phase; Polymers; Regimen; Route; Salmonella; Salmonella enterica; Salmonella typhimurium; Scanning Electron Microscopy; Solubility; Spleen; Time; Tissues; Tuberculosis; Tularemia; Typhoid Fever; Vaccines; Virulent; bactericide; celecoxib; combat; cytokine; dextran; dosage; efficacy evaluation; immunotoxicity; in vivo; inhibitor/antagonist; macrophage; monocyte; nanoparticle; novel; oral infection; particle; pathogen; prevent; small molecule

DESCRIPTION (provided by applicant): AR-12 is an IND approved, COX-2 inhibitor derived drug that affects pathogen host cells by primarily upregulating autophagy. In vitro, AR-12 has shown broad spectrum efficacy on several bacterial strains including S. typhimurium, F. tularensis (Schu S4, LVS), and F. novicida. In vivo, AR-12 given i.v. reduced organ bacterial tenfold compared to untreated controls, but did not prevent host death due to typhoid fever. AR-12 concentrations were limited with in vivo application because the of the drug's hydrophobicity. To overcome solubility issues, we propose encapsulating AR-12 in acetalated dextran (Ac-DEX) particles that passively target the host cell. Ac-DEX is an acid sensitive polymer with tunable release kinetics that will release drug in the phagocyte's phagosome, due to the lower pH present. There are three aims for the R21 portion of this proposal. The first aim is to manufacture and characterize two types of Ac-DEX particles that encapsulate AR- 12: 1) a nanoparticle (NP) for i.v. or i.p. injection that is at the ideal size for passively targeting macrophages (500-1,000 nm); 2) a porous microparticle (PMP) that is ideal size (5-15 mm) for inhalation via the nose (i.n.) and penetration to the nasal associated lymphoid tissue (NALT). These particles will be manufactured, imaged through scanning electron microscopy, and characterized for drug loading through fluorescence spectroscopy. The second aim is to evaluate the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of unencapsulated AR-12 against drug susceptible and multidrug resistant M. tuberculosis (MDR TB) and encapsulated AR-12 against TB, F. tularensis (Schu S4), and S. typhimurium in human macrophages. Macrophages will be infected with one of three bacteria and treated with AR-12. Cell associated bacteria and CFUs will be evaluated to determine both the MIC and MBC. The MIC and MBC determined for F. tularensis will be used in Aim 3 for in vivo treatment with encapsulated AR-12 administered i.v. and i.p. with NPs, and i.n. with PMPs. A dosing study will be used to identify the best route and dosage (MIC, MBC, and 10 x MBC) by evaluating organ CFUs when administered at a comparable timing and frequency to traditional antibiotic regimens. The optimum route and dose will be used for an in-depth treatment evaluation of organ CFUs, histopathology, blood cytokine levels and survival. The milestones for progress to the R33 portion of the grant will be 1) encapsulation of AR-12 in Ac-DEX particles; 2) reduced macrophage associated M. tuberculosis with AR-12 treatment; 3) Increased survival and decreased organ bacterial load with AR-12 treatment of F. tularensis, in vivo. The R33 phase of this grant also has 3 specific aims. Aim 4 and 5 are to evaluate encapsulated AR-12 treatment in vivo against S. typhimurium, and TB, respectively in a manner similar to Aim 3. Aim 6 is to evaluate the immunotoxicity of encapsulated AR-12 and to progress encapsulated AR-12 towards IND approval. These studies will help to develop and characterize a new broad spectrum antibiotic and delivery platform that targets host cells. PUBLIC HEALTH RELEVANCE: Therapies that target the host cell rather than the bacteria, which traditional antibiotics primarily focus, can help to combat pathogens that have drug resistance. AR-12 is an antibiotic that has shown broad spectrum activity against multiple bacterial pathogens by primarily effecting the infected host cell. We propose to further evaluate AR-12 in animal models against bacteria that cause salmonella, tularemia, drug susceptible tuberculosis and multi-drug resistant tuberculosis.

描述（由申请人提供）：AR-12 是一种 IND 批准的 COX-2 抑制剂衍生药物，主要通过上调自噬来影响病原体宿主细胞。在体外，AR-12 对多种细菌菌株显示出广谱功效，包括鼠伤寒沙门氏菌、土拉杆菌 (Schu S4、LVS) 和新杀杆菌。在体内，AR-12 静脉注射。与未经治疗的对照组相比，器官细菌减少了十倍，但并不能阻止宿主因伤寒而死亡。由于药物的疏水性，AR-12 的浓度在体内应用中受到限制。为了克服溶解度问题，我们建议将 AR-12 封装在被动靶向宿主细胞的乙酰化葡聚糖 (Ac-DEX) 颗粒中。 Ac-DEX 是一种酸敏感聚合物，具有可调节的释放动力学，由于存在较低的 pH 值，它将在吞噬细胞的吞噬体中释放药物。该提案的 R21 部分有三个目标。第一个目标是制造和表征两种类型的封装 AR-12 的 Ac-DEX 颗粒：1) 用于静脉注射的纳米颗粒 (NP)。或IP注射尺寸适合被动靶向巨噬细胞（500-1,000 nm）； 2) 多孔微粒 (PMP)，尺寸理想（5-15 毫米），适合通过鼻子吸入 (i.n) 并渗透到鼻相关淋巴组织 (NALT)。这些颗粒将被制造，通过扫描电子显微镜成像，并通过荧光光谱法表征药物负载。第二个目的是评估未封装的 AR-12 对药物敏感和多重耐药结核分枝杆菌 (MDR TB) 的最低抑菌浓度 (MIC) 和最低杀菌浓度 (MBC) 以及封装的 AR-12 对人类巨噬细胞中的结核菌、土拉杆菌 (Schu S4) 和鼠伤寒沙门氏菌的最低抑菌浓度 (MIC) 和最低杀菌浓度 (MBC)。巨噬细胞将被三种细菌中的一种感染并用 AR-12 处理。将评估细胞相关细菌和 CFU，以确定 MIC 和 MBC。土拉弗朗西斯测定的 MIC 和 MBC 将用于目标 3，通过静脉内施用封装的 AR-12 进行体内治疗。和IP。与 NP 和 i.n.与 PMP 一起。剂量研究将用于通过评估以与传统抗生素治疗方案相当的时间和频率施用时的器官 CFU 来确定最佳途径和剂量（MIC、MBC 和 10 x MBC）。最佳途径和剂量将用于对器官CFU、组织病理学、血液细胞因子水平和存活率进行深入的治疗评估。拨款 R33 部分进展的里程碑将是 1) 将 AR-12 封装在 Ac-DEX 颗粒中； 2) AR-12治疗减少了与结核分枝杆菌相关的巨噬细胞； 3) AR-12 处理土拉弗拉菌体内可提高存活率并降低器官细菌负荷。这笔赠款的 R33 阶段还有 3 个具体目标。目标 4 和 5 是以类似于目标 3 的方式分别评估封装的 AR-12 在体内针对鼠伤寒沙门氏菌和结核病的治疗。目标 6 是评估封装的 AR-12 的免疫毒性并推动封装的 AR-12 获得 IND 批准。这些研究将有助于开发和表征一种针对宿主细胞的新型广谱抗生素和递送平台。 公共卫生相关性：针对宿主细胞而不是传统抗生素主要针对细菌的疗法可以帮助对抗具有耐药性的病原体。 AR-12 是一种抗生素，通过主要影响受感染的宿主细胞，对多种细菌病原体表现出广谱活性。我们建议在动物模型中进一步评估 AR-12 对引起沙门氏菌、兔热病、药物敏感结核病和多重耐药结核病的细菌的作用。