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: 8910618
• 负责人: Kristy M Ainslie
• 金额: $ 31.53万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8910618
• 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; dosage; efficacy evaluation; immunotoxicity; in vivo; inhibitor/antagonist; macrophage; monocyte; nanoparticle; novel; oral infection; particle; pathogen; prevent; small molecule; targeted treatment

Project Summary AR-12 is an IND approved, COX-2 inhibitor derived drug that affects pathogen host cells by primarily up- regulating 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 10-fold 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 3 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 m) 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, 10xMBC) 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.

项目摘要 AR-12是由IND批准的COX-2抑制剂衍生的药物 调节自噬。在体外，AR-12在几种细菌菌株中显示出广泛的效率 S. Typhimurium，F。tularensis（Schu S4，LVS）和F. Novicida。在体内，AR-12给定i.v.减少器官 与未处理的对照相比，细菌10倍，但由于伤寒而导致宿主死亡。 AR-12 由于药物的疏水性，体内施用的浓度受到限制。克服 溶解度问题，我们提出在乙酰化葡萄糖（AC-DEX）颗粒中封装AR-12 靶向宿主单元。 AC-DEX是一种具有可调释放动力学的酸敏感聚合物，将在 由于存在较低的pH，吞噬细胞的吞噬体。 R21部分有3个目标 提议。第一个目的是制造和表征两种封装Ar-的AC-DEX颗粒 12：1）i.v.的纳米颗粒（NP）或i.p.被动靶向巨噬细胞的理想大小的注射 （500-1,000 nm）; 2）用于通过鼻子吸入的理想大小（5-15 m）的多孔微粒（PMP）（I.N.） 并渗透到鼻相关淋巴组织（NALT）。这些粒子将是制造的，成像 通过扫描电子显微镜，并通过荧光光谱进行药物载荷进行表征。 第二个目的是评估最小抑制浓度（MIC）和最小杀菌性 未包容的AR-12浓度（MBC）针对药物易感和多药耐药。 结核病（MDR TB）和针对TB，F。tularensis（Schu S4）和鼠伤寒S. 人类巨噬细胞。巨噬细胞将被三种细菌之一感染并用AR-12治疗。细胞 将评估相关的细菌和CFU，以确定MIC和MBC。麦克风和MBC 针对F. tularensis确定的AIM 3将使用封装的AR-12进行体内治疗 I.V.和I.P.与NP和I.N.与PMP。剂量研究将用于识别最佳路线和剂量 （MIC，MBC，10XMBC）通过评估器官CFUS以类似的时间和频率给药 传统的抗生素方案。最佳路线和剂量将用于深度治疗评估 器官CFU，组织病理学，血细胞因子水平和生存。 R33进度的里程碑 赠款的一部分将为1）AR-12在AC-DEX颗粒中的封装； 2）减少巨噬细胞相关 链球菌用AR-12治疗进行结核； 3）增加生存率并改善了AR-12的有机细菌负荷 F. tularensis的治疗，体内。该赠款的R33阶段还具有3个具体目标。目标4和5是 分别以某种方式评估了针对伤寒链霉菌和TB的封装AR-12处理。 与目标3相似。目标6是评估封装AR-12的免疫毒性并进行封装的进展 AR-12获得IND批准。这些研究将有助于发展和表征新的广泛范围 靶向宿主细胞的抗生素和输送平台。