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抑制剂衍生的药物，主要通过上调自噬来影响病原体宿主细胞。在体外，AR-12在几种细菌菌株中显示出广泛的频谱功效，包括鼠伤寒沙门氏菌，tularensis（Schu S4，LVS）和F. novicida。在体内，AR-12给定i.v.与未经治疗的对照相比，器官细菌的十倍减少，但由于伤寒引起的宿主死亡并不能阻止宿主死亡。由于药物的疏水性，AR-12浓度受体内施用的限制。为了克服溶解度问题，我们提出将AR-12封装在被动针对宿主细胞的乙酰化葡萄糖（AC-DEX）颗粒中。 AC-DEX是一种具有可调释放动力学的酸敏感聚合物，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。第一个目的是制造和表征两种封装AR-12：1）i.v的纳米颗粒（NP）的AC-脱核颗粒。或i.p.被动靶向巨噬细胞（500-1,000 nm）的理想大小的注射； 2）一个是理想大小（5-15 mm）的多孔微粒（PMP），可通过鼻子（I.N.）吸入并穿透到鼻相关淋巴组织（NALT）。这些颗粒将通过扫描电子显微镜制成，成像，并通过荧光光谱进行药物加载。第二个目的是评估未封装AR-12的最低抑制浓度（MIC）和最低杀菌浓度（MBC），以抗药物易感和耐多药的结核分枝杆菌（MDR TB），并封装了针对TB，F。tularensis（Shu s4）（Schu S4）和S. typhimophium的AR-12。巨噬细胞将被三种细菌之一感染并用AR-12治疗。将评估与细胞相关的细菌和CFU，以确定MIC和MBC。针对F. tularensis确定的MIC和MBC将用于AIM 3用于体内治疗，并用封装的AR-12施用的i.v.和I.P.与NP和I.N.与PMP。剂量研究将用于确定最佳的途径和剂量（MIC，MBC和10 X MBC），通过评估在与传统抗生素方案的可比时间和频率下进行施用时，通过评估器官CFU。最佳途径和剂量将用于对器官CFU，组织病理学，血细胞因子水平和存活的深入治疗评估。赠款的R33部分的里程碑将为1）AC-DEX颗粒中AR-12的封装； 2）通过AR-12处理减少巨噬细胞的结核分枝杆菌； 3）体内的AR-12处理f. tularensis的AR-12处理增加了生存率和细菌载荷。该赠款的R33阶段还具有3个具体目标。 AIM 4和5分别以类似于AIM 3的方式评估对鼠伤寒沙门氏菌和TB的体内封装的AR-12处理。AIM6是评估封装AR-12的免疫毒性，并进步封装的AR-12在IND批准方面进行封装的AR-12。这些研究将有助于开发和表征针对宿主细胞的新型广谱抗生素和输送平台。 公共卫生相关性：靶向宿主细胞而不是传统抗生素主要集中的细菌的疗法可以帮助打击具有耐药性的病原体。 AR-12是一种抗生素，它通过主要影响感染的宿主细胞表现出针对多种细菌病原体的广泛谱活性。我们建议在动物模型中进一步评估AR-12，以针对细菌，导致沙门氏菌，tular症，药物易感结核病和多药耐药性结核病。