Overcoming Pyrazinamide Resistance with Pyrazinoate-Cephalosporin Conjugates

用吡嗪酸-头孢菌素缀合物克服吡嗪酰胺耐药性

• 批准号: 10088387
• 负责人: Courtney C Aldrich
• 金额: $ 19.17万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-25 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10088387
• 关键词: AIDS-Related Opportunistic Infections; Atypical Mycobacteria; Bacillus; Basic Science; Bioavailable; Biochemical; Caco-2 Cells; Cephalosporins; Clinical; Communicable Diseases; Data; Development; Drug Kinetics; Drug resistance; Enzymes; Esters; Etiology; Future; Genetic; Human; Immunology; In Vitro; Infection; Infectious Diseases Research; Liquid substance; Measures; Mediating; Microbiology; Minnesota; Modeling; Mus; Mycobacterium tuberculosis; Oral; Patients; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Population; Predisposition; Prodrugs; Pyrazinamide; Pyrazinamide resistance; Relapse; Research; Resistance; Serum; Site; Stomach; Treatment Failure; Treatment Protocols; Tuberculosis; Universities; Work; amidase; analog; antimicrobial; beta-Lactamase; beta-Lactams; collaborative environment; commensal microbes; drug discovery; drug disposition; esterase; gut microbiome; in vivo; loss of function mutation; macrophage; microorganism; mortality; mycobacterial; novel; pathogen; preclinical development; pyrazinoic acid; research facility; therapy duration; tuberculosis chemotherapy; tuberculosis drugs; tuberculosis treatment

SUMMARY Mycobacterium tuberculosis (Mtb), the principal etiological agent of tuberculosis (TB), infects over one-third of humanity and is now the leading cause of infectious disease mortality by a single pathogen. Pyrazinamide (PZA) is a first-line sterilizing anti-tubercular drug that is anticipated to be an irreplaceable component of future TB treatment regimens. Previous studies have demonstrated that PZA is a pro-drug which is converted to the active form of pyrazinoic acid (POA) by the M. tuberculosis amidase PncA, and that loss of function mutations in pncA account for the vast majority of PZA resistance. Resistance to PZA in turn is associated with significantly higher treatment failures and relapse rates. Attempts to directly administer POA in humans and mice have been unsuccessful because POA cannot attain sufficient concentrations within Mtb at the infection site. Prodrug approaches employing simple POA esters have been unable to overcome the intrinsic drug disposition liabilities of POA as conventional prodrugs are rapidly hydrolyzed by serum esterases. In preliminary data, we have developed a novel prodrug strategy for POA through conjugation to a β-lactam promoiety leading to selective release of POA by the unique mycobacterial beta-lactamase BlaC and not by serum esterases. This strategy overcomes PZA-resistance by circumventing PncA-mediated activation and renders blaC conditionally essential since loss of function mutations to blaC, restores susceptibility of Mtb to the β-lactam promoiety. Consequently, we hypothesize the genetic barrier for development of spontaneous resistance to our POA-β-lactam conjugates will be high. The objectives of this application are to synthesize and evaluate β-lactam prodrugs of POA, which are orally bioavailable and selectively release POA within Mtb- infected macrophages. We will accomplish the overall objectives of this application by pursuing two specific aims. In aim, we will synthesize β-lactam prodrugs of POA, which are selectively activated by Mtb to release POA and not by commensal microbiota. The prodrugs will be biochemically and microbiologically characterized with a panel of β-lactamases and microorganisms. In aim 2, we will measure prodrug stability in serum and stimulated gastric fluid and permeability in Caco-2 cells. We will next determine complete pharmacokinetic parameters and the extent of POA release by the gut microbiome. Finally, we plan to evaluate one compound in a murine TB infection model.

概括 结核分枝杆菌 (Mtb) 是结核病 (TB) 的主要病原体，感染超过三分之一的人 人类，现在是由单一病原体引起的传染病死亡的主要原因。 （PZA）是一线灭菌抗结核药物，预计将成为未来不可替代的组成部分 以前的研究表明，PZA 是一种前药，可转化为结核病治疗方案。 结核分枝杆菌酰胺酶 PncA 产生吡嗪酸 (POA) 的活性形式，以及功能缺失突变 pncA 占 PZA 耐药的绝大多数，而 PZA 耐药又与此相关。 尝试在人类中直接施用 POA 的治疗失败率和复发率明显更高。 小鼠未能成功，因为感染时 POA 在 Mtb 内无法达到足够的浓度 使用简单的 POA 酯的前药方法无法克服内在药物。 POA 作为常规前药的处置倾向会被血清酯酶快速水解。 根据初步数据，我们通过与 β-内酰胺缀合开发了 POA 的新型前药策略 促进部分导致 POA 通过独特的分枝杆菌 β-内酰胺酶 BlaC 选择性释放，而不是通过 该策略通过规避 PncA 介导的激活来克服 PZA 耐药性。 使 blaC 有条件地必需，因为 blaC 的功能突变丧失，恢复 Mtb 对 测试了β-内酰胺基团，我们克服了自发发育的遗传障碍。 对我们的 POA-β-内酰胺缀合物的耐药性将很高 该应用的目标是合成和。 评估 POA 的 β-内酰胺前药，这些药物具有口服生物利用度，并在 Mtb- 内选择性释放 POA 我们将通过追求两个特定的目标来实现该应用的总体目标。 目标是合成POA的β-内酰胺前药，其被Mtb选择性激活释放。 POA 而不是通过共生微生物群对前药进行生化和微生物学表征。 在目标 2 中，我们将测量血清和微生物中的前药稳定性。 接下来我们将确定完整的药代动力学。 最后，我们计划评估一种化合物。 在小鼠结核感染模型中。

项目成果

Cephem-Pyrazinoic Acid Conjugates: Circumventing Resistance in Mycobacterium tuberculosis.

头孢烯-吡嗪酸缀合物：规避结核分枝杆菌的耐药性。

• 发表时间: 2022-09-12
• 作者: Cole, Malcolm S;Howe, Michael D;Buonomo, Joseph A;Sharma, Sachin;Lamont, Elise A;Brody, Scott I;Mishra, Neeraj K;Minato, Yusuke;Thiede, Joshua M;Baughn, Anthony D;Aldrich, Courtney C
• 通讯作者: Aldrich, Courtney C