Investigation of synergy between Rifampin and SQ109, a new anti-TB drug candidate

利福平与新型抗结核候选药物SQ109之间的协同作用研究

• 批准号: 7925148
• 负责人: VENKATA M REDDY
• 金额: $ 69.35万
• 依托单位: SEQUELLA, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-22 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7925148
• 关键词: Adverse effects; Anabolism; Animals; Antibiotics; Antitubercular Agents; Applications Grants; Area; Biological; Biological Assay; Biotechnology; Cell Wall; Characteristics; Chemicals; Clinical Trials; Collaborations; Combination Drug Therapy; Combined Modality Therapy; Cytochrome P450; Data; Developed Countries; Developing Countries; Development; Diamines; Disease; Dose; Drug Compounding; Drug Interactions; Drug Kinetics; Drug Metabolic Detoxication; Drug resistance in tuberculosis; Drug toxicity; Enzymes; Ethambutol; Failure; Fatty Acids; Frequencies; Future; Genes; Genetic Transcription; Genus Mycobacterium; Grant; Growth; HIV; Heme; Human; Human Volunteers; Immunosuppression; In Vitro; Individual; Infection; Investigation; Knock-out; Knowledge; Lead; Libraries; Link; Liver Microsomes; Lung; Malnutrition; Mammalian Cell; Manuscripts; Mass Spectrum Analysis; Mediating; Metabolism; Modeling; Multi-Drug Resistance; Multidrug-Resistant Tuberculosis; Mus; Mycobacterium tuberculosis; Oral; Pharmaceutical Preparations; Pharmacodynamics; Phase; Play; Principal Investigator; Production; Property; Proteins; Pulmonary Tuberculosis; Pyrazinamide; Reaction; Respiratory System; Respiratory tract structure; Rifampin; Role; Steroids; Sterols; Structure; Testing; Therapeutic; Time; Tissues; Treatment Protocols; Tuberculosis; Work; World Health Organization; Xenobiotics; bactericide; combat; cytotoxicity; design; drug candidate; drug standard; in vitro Assay; in vivo; interest; isoniazid; killings; macrophage; mortality; mouse model; mutant; non-compliance; novel diagnostics; pre-clinical; preclinical study; prevent; programs; public health relevance; research and development; research study; resistant strain; tool; treatment duration; tuberculosis drugs; tuberculosis treatment

DESCRIPTION (provided by applicant): SQ109 is a new anti-TB drug candidate that is currently undergoing Phase Ia Clinical Trials in humans. SQ109 was selected out of the library of 63,238 compounds, because it is highly efficacious in killing M. tuberculosis within infected macrophages and in experimental animals, has relatively low cytotoxicity in cultured mammalian cells, has activity against both drug-susceptible and drug-resistant M. tuberculosis, and has drug-like pharmacokinetic and pharmacodynamic profiles. Any new anti-TB drug will be used in combination with other drugs in order to prevent the emergence of drug resistant M. tuberculosis. How SQ109, a new drug compound with unique attributes, interacts with existing first line anti-TB drugs is crucial for us to understand, as this knowledge enables us to design the most efficacious therapeutic combination and to avoid any antagonistic side effects of a new combination therapy. We investigated the drug interaction in various combinations of SQ109 with the first line anti-TB drugs in both in vitro assays and in vivo murine models of TB. We found that SQ109 preferentially synergized with RIF in in vitro growth inhibition assays and demonstrated enhanced bactericidal activity in mice when used in combination with INH and RIF. These findings are significant for optimizing the drug combination therapies to be tested in future clinical trials. In this exploratory R21 grant Application we will: 1. Assess whether SQ109 is metabolized within M. tuberculosis and how co-administration of RIF influences SQ109 metabolism. If SQ109 metabolites exist, we will study their structures by mass spectrometry and will compare them to the SQ109 metabolites generated by human CYP. If the metabolites are the same, we will prepare and purify them in sufficient quantities and test them against M. tuberculosis for potency in vitro. 2. Determine the expression of individual CYP in RIF-treated M. tuberculosis in order to identify candidate SQ109 activators that are induced by RIF. 3. Generate M. tuberculosis knock out mutants of the candidate CYP(s) and re-examine the synergy of interaction between SQ109 and RIF in the CYP knockout mutants. Public Health Relevance: One of the most vexing problems with standard tuberculosis (TB) therapy is the lengthy treatment course (6-9 months) and toxicity of the drugs in the regiment. This prolonged therapeutic regimen results in a high level of noncompliance, and ineffective concentrations of drugs lead to the emergence of multidrug resistant (MDR) M. tuberculosis. There are an estimated 400,000 new cases of MDR TB each year, and the World Health Organization (WHO) estimates there are 50 million individuals worldwide infected with a resistant strain of TB. MDR strains of M. tuberculosis cause increased mortality, and the fatality rate of untreated cases of MDR TB can be as high as 70%. The likelihood of cure for MDR M. tuberculosis-infected individuals becomes even less in the setting of secondary immunosuppression caused by HIV and malnutrition, conditions that are rampant in developing countries. Because of the failures of the existing therapies, the world desperately needs new drugs that can shorten treatment duration and kill MDR M. tuberculosis. Sequella, Inc., founded in 1997, focuses its R&D on developing new diagnostic tools and new drugs for TB disease. One of our most successful projects is the development of SQ109, a new anti-TB drug candidate that is currently undergoing Phase Ia Clinical Trials. SQ109 was selected as our lead drug candidate out of the library of 63,238 compounds, because it is highly efficacious in killing M. tuberculosis within infected macrophages and in experimental animals, has relatively low cytotoxicity in cultured mammalian cells, has activity against both drug-susceptible and drug-resistant M. tuberculosis, and has drug-like pharmacokinetic and pharmacodynamic profiles. Importantly, when used in combination with rifampin (RIF) + isoniazid (INH) + pyrazinamide (PZA), SQ109 is able to increase bacterial killing and reduce the time by at least 25% to achieve the same level of CFU as the standard drug regimen, RIF + INH + PZA + ethambutol (EMB) in a mouse model of TB. Any new anti-TB drug will be used in combination with other drugs in order to prevent the emergence of drug resistant M. tuberculosis. How SQ109, a new drug compound with unique attributes, interacts with existing first line anti-TB drugs is crucial for us to understand, as this knowledge enables us to design the most efficacious therapeutic combination and to avoid any antagonistic side effects of a new combination therapy. In this R21 exploratory grant application, we propose to investigate one of the several hypotheses that may contribute to the synergistic interaction between SQ109 and RIF in M. tuberculosis: (1) SQ109 is readily metabolized in M. tuberculosis, (2) RIF is able to induce the expression of one or more M. tuberculosis CYP- like proteins, and (3) CYP induced by RIF is involved in SQ109 metabolism. Tasks 1 and 2 are two independent areas of study, which are linked by task 3. In this application, we outline the experiment design for all three tasks in sequence. Further, we provide alternative directions for the study of this interesting synergy if the data obtained do not support our hypothesis.

描述（由申请人提供）：SQ109是一种新的抗结核候选药物，目前正在进行人体Ia期临床试验。 SQ109是从63,238个化合物库中选出的，因为它在杀死受感染的巨噬细胞和实验动物中的结核分枝杆菌方面非常有效，在培养的哺乳动物细胞中具有相对较低的细胞毒性，对药物敏感和耐药的结核分枝杆菌均​​具有活性，并且具有类似药物的药代动力学和药效学特征。任何新的抗结核药物都会与其他药物联合使用，以防止耐药结核分枝杆菌的出现。 SQ109这种具有独特属性的新药物化合物如何与现有的一线抗结核药物相互作用对于我们来说至关重要，因为这些知识使我们能够设计出最有效的治疗组合，并避免新组合疗法的任何拮抗副作用。我们在结核病的体外测定和体内小鼠模型中研究了 SQ109 与一线抗结核药物的各种组合的药物相互作用。我们发现，在体外生长抑制试验中，SQ109 优先与 RIF 协同作用，并且与 INH 和 RIF 联合使用时，在小鼠中表现出增强的杀菌活性。这些发现对于优化未来临床试验中待测试的药物组合疗法具有重要意义。在此探索性 R21 资助申请中，我们将： 1. 评估 SQ109 是否在结核分枝杆菌内代谢，以及 RIF 的共同给药如何影响 SQ109 代谢。如果 SQ109 代谢物存在，我们将通过质谱研究其结构，并将其与人类 CYP 产生的 SQ109 代谢物进行比较。如果代谢物相同，我们将制备和纯化足够数量的它们，并在体外测试它们对结核分枝杆菌的效力。 2. 确定 RIF 处理的结核分枝杆菌中单个 CYP 的表达，以鉴定 RIF 诱导的候选 SQ109 激活剂。 3.生成候选CYP的结核分枝杆菌敲除突变体，并重新检查CYP敲除突变体中SQ109和RIF之间相互作用的协同作用。 公共卫生相关性：标准结核病 (TB) 治疗最令人烦恼的问题之一是疗程较长（6-9 个月）和药物毒性。这种延长的治疗方案会导致高度的不依从性，并且无效的药物浓度会导致耐多药（MDR）结核分枝杆菌的出现。据估计，每年新增耐多药结核病例 40 万例，世界卫生组织 (WHO) 估计全球有 5000 万人感染耐药结核菌株。耐多药结核分枝杆菌菌株导致死亡率增加，未经治疗的耐多药结核病病例的死亡率可高达 70%。在艾滋病毒和营养不良（发展中国家普遍存在的情况）引起的继发性免疫抑制的情况下，耐多药结核分枝杆菌感染者的治愈可能性甚至更低。 由于现有疗法的失败，世界迫切需要能够缩短治疗时间并杀死耐多药结核分枝杆菌的新药。 Sequella, Inc. 成立于 1997 年，其研发重点是开发结核病的新诊断工具和新药物。我们最成功的项目之一是 SQ109 的开发，这是一种新的抗结核候选药物，目前正在进行 Ia 期临床试验。 SQ109从63,238个化合物库中被选为我们的主要候选药物，因为它在杀死受感染的巨噬细胞和实验动物中的结核分枝杆菌方面非常有效，在培养的哺乳动物细胞中具有相对较低的细胞毒性，对药物敏感和耐药的结核分枝杆菌均​​具有活性，并且具有类似药物的药代动力学和药效学特征。重要的是，当与利福平 (RIF) + 异烟肼 (INH) + 吡嗪酰胺 (PZA) 联合使用时，SQ109 能够提高细菌杀灭率，并缩短至少 25% 的时间，以在结核病小鼠模型中达到与标准药物方案 RIF + INH + PZA + 乙胺丁醇 (EMB) 相同的 CFU 水平。 任何新的抗结核药物都会与其他药物联合使用，以防止耐药结核分枝杆菌的出现。 SQ109这种具有独特属性的新药物化合物如何与现有的一线抗结核药物相互作用对于我们来说至关重要，因为这些知识使我们能够设计出最有效的治疗组合，并避免新组合疗法的任何拮抗副作用。 在此 R21 探索性资助申请中，我们建议研究可能有助于 SQ109 和 RIF 在结核分枝杆菌中协同相互作用的几个假设之一：（1）SQ109 在结核分枝杆菌中很容易代谢，（2）RIF 能够诱导一种或多种结核分枝杆菌 CYP 样蛋白的表达，（3）RIF 诱导的 CYP 参与 SQ109新陈代谢。任务 1 和 2 是两个独立的研究领域，通过任务 3 联系起来。在此应用程序中，我们按顺序概述了所有三个任务的实验设计。此外，如果获得的数据不支持我们的假设，我们将为研究这种有趣的协同作用提供替代方向。