Mary Nantongo, David C. Nguyen, Christopher R. Bethel, Magdalena A. Taracila, Qing Li, Khalid M. Dousa, Eunjeong Shin, Sebastian G. Kurz, Liem Nguyen, Barry N. Kreiswirth, W. Henry Boom, Mark S. Plummer and Robert A. Bonomo*,
{"title":"双氮双环辛烷 β-内酰胺酶抑制剂杜洛巴坦能抑制结核分枝杆菌的 BlaC 和肽聚糖转肽酶","authors":"Mary Nantongo, David C. Nguyen, Christopher R. Bethel, Magdalena A. Taracila, Qing Li, Khalid M. Dousa, Eunjeong Shin, Sebastian G. Kurz, Liem Nguyen, Barry N. Kreiswirth, W. Henry Boom, Mark S. Plummer and Robert A. Bonomo*, ","doi":"10.1021/acsinfecdis.4c00119","DOIUrl":null,"url":null,"abstract":"<p >Peptidoglycan synthesis is an underutilized drug target in <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>). Diazabicyclooctanes (DBOs) are a class of broad-spectrum β-lactamase inhibitors that also inhibit certain peptidoglycan transpeptidases that are important in mycobacterial cell wall synthesis. We evaluated the DBO durlobactam as an inhibitor of BlaC, the <i>Mtb</i> β-lactamase, and multiple <i>Mtb</i> peptidoglycan transpeptidases (PonA1, Ldt<sub>Mt1</sub>, Ldt<sub>Mt2</sub>, Ldt<sub>Mt3</sub>, and Ldt<sub>Mt5</sub>). Timed electrospray ionization mass spectrometry (ESI-MS) captured acyl-enzyme complexes with BlaC and all transpeptidases except Ldt<sub>Mt5</sub>. Inhibition kinetics demonstrated durlobactam was a potent and efficient DBO inhibitor of BlaC (<i>K</i><sub>I app</sub> 9.2 ± 0.9 μM, <i>k</i><sub>2</sub>/<i>K</i> 5600 ± 560 M<sup>–1</sup> s<sup>–1</sup>) and similar to clavulanate (<i>K</i><sub>I app</sub> 3.3 ± 0.6 μM, <i>k</i><sub>2</sub>/<i>K</i> 8400 ± 840 M<sup>–1</sup> s<sup>–1</sup>); however, durlobactam had a lower turnover number (<i>t</i><sub>n</sub> = <i>k</i><sub>cat</sub>/<i>k</i><sub>inact</sub>) than clavulanate (1 and 8, respectively). <i>K</i><sub>I app</sub> values with durlobactam and clavulanate were similar for peptidoglycan transpeptidases, but ESI-MS captured durlobactam complexes at more time points. Molecular docking and simulation demonstrated several productive interactions of durlobactam in the active sites of BlaC, PonA1, and Ldt<sub>Mt2</sub>. Antibiotic susceptibility testing was conducted on 11 <i>Mtb</i> isolates with amoxicillin, ceftriaxone, meropenem, imipenem, clavulanate, and durlobactam. Durlobactam had a minimum inhibitory concentration (MIC) range of 0.5–16 μg/mL, similar to the ranges for meropenem (1–32 μg/mL) and imipenem (0.5–64 μg/mL). In β-lactam + durlobactam combinations (1:1 mass/volume), MICs were lowered 4- to 64-fold for all isolates except one with meropenem–durlobactam. This work supports further exploration of novel β-lactamase inhibitors that target BlaC and <i>Mtb</i> peptidoglycan transpeptidases.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":4.0000,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Durlobactam, a Diazabicyclooctane β-Lactamase Inhibitor, Inhibits BlaC and Peptidoglycan Transpeptidases of Mycobacterium tuberculosis\",\"authors\":\"Mary Nantongo, David C. Nguyen, Christopher R. Bethel, Magdalena A. Taracila, Qing Li, Khalid M. Dousa, Eunjeong Shin, Sebastian G. Kurz, Liem Nguyen, Barry N. Kreiswirth, W. Henry Boom, Mark S. Plummer and Robert A. Bonomo*, \",\"doi\":\"10.1021/acsinfecdis.4c00119\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p >Peptidoglycan synthesis is an underutilized drug target in <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>). Diazabicyclooctanes (DBOs) are a class of broad-spectrum β-lactamase inhibitors that also inhibit certain peptidoglycan transpeptidases that are important in mycobacterial cell wall synthesis. We evaluated the DBO durlobactam as an inhibitor of BlaC, the <i>Mtb</i> β-lactamase, and multiple <i>Mtb</i> peptidoglycan transpeptidases (PonA1, Ldt<sub>Mt1</sub>, Ldt<sub>Mt2</sub>, Ldt<sub>Mt3</sub>, and Ldt<sub>Mt5</sub>). Timed electrospray ionization mass spectrometry (ESI-MS) captured acyl-enzyme complexes with BlaC and all transpeptidases except Ldt<sub>Mt5</sub>. Inhibition kinetics demonstrated durlobactam was a potent and efficient DBO inhibitor of BlaC (<i>K</i><sub>I app</sub> 9.2 ± 0.9 μM, <i>k</i><sub>2</sub>/<i>K</i> 5600 ± 560 M<sup>–1</sup> s<sup>–1</sup>) and similar to clavulanate (<i>K</i><sub>I app</sub> 3.3 ± 0.6 μM, <i>k</i><sub>2</sub>/<i>K</i> 8400 ± 840 M<sup>–1</sup> s<sup>–1</sup>); however, durlobactam had a lower turnover number (<i>t</i><sub>n</sub> = <i>k</i><sub>cat</sub>/<i>k</i><sub>inact</sub>) than clavulanate (1 and 8, respectively). <i>K</i><sub>I app</sub> values with durlobactam and clavulanate were similar for peptidoglycan transpeptidases, but ESI-MS captured durlobactam complexes at more time points. Molecular docking and simulation demonstrated several productive interactions of durlobactam in the active sites of BlaC, PonA1, and Ldt<sub>Mt2</sub>. Antibiotic susceptibility testing was conducted on 11 <i>Mtb</i> isolates with amoxicillin, ceftriaxone, meropenem, imipenem, clavulanate, and durlobactam. Durlobactam had a minimum inhibitory concentration (MIC) range of 0.5–16 μg/mL, similar to the ranges for meropenem (1–32 μg/mL) and imipenem (0.5–64 μg/mL). In β-lactam + durlobactam combinations (1:1 mass/volume), MICs were lowered 4- to 64-fold for all isolates except one with meropenem–durlobactam. This work supports further exploration of novel β-lactamase inhibitors that target BlaC and <i>Mtb</i> peptidoglycan transpeptidases.</p>\",\"PeriodicalId\":17,\"journal\":{\"name\":\"ACS Infectious Diseases\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.0000,\"publicationDate\":\"2024-04-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Infectious Diseases\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://pubs.acs.org/doi/10.1021/acsinfecdis.4c00119\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, MEDICINAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Infectious Diseases","FirstCategoryId":"3","ListUrlMain":"https://pubs.acs.org/doi/10.1021/acsinfecdis.4c00119","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, MEDICINAL","Score":null,"Total":0}
Durlobactam, a Diazabicyclooctane β-Lactamase Inhibitor, Inhibits BlaC and Peptidoglycan Transpeptidases of Mycobacterium tuberculosis
Peptidoglycan synthesis is an underutilized drug target in Mycobacterium tuberculosis (Mtb). Diazabicyclooctanes (DBOs) are a class of broad-spectrum β-lactamase inhibitors that also inhibit certain peptidoglycan transpeptidases that are important in mycobacterial cell wall synthesis. We evaluated the DBO durlobactam as an inhibitor of BlaC, the Mtb β-lactamase, and multiple Mtb peptidoglycan transpeptidases (PonA1, LdtMt1, LdtMt2, LdtMt3, and LdtMt5). Timed electrospray ionization mass spectrometry (ESI-MS) captured acyl-enzyme complexes with BlaC and all transpeptidases except LdtMt5. Inhibition kinetics demonstrated durlobactam was a potent and efficient DBO inhibitor of BlaC (KI app 9.2 ± 0.9 μM, k2/K 5600 ± 560 M–1 s–1) and similar to clavulanate (KI app 3.3 ± 0.6 μM, k2/K 8400 ± 840 M–1 s–1); however, durlobactam had a lower turnover number (tn = kcat/kinact) than clavulanate (1 and 8, respectively). KI app values with durlobactam and clavulanate were similar for peptidoglycan transpeptidases, but ESI-MS captured durlobactam complexes at more time points. Molecular docking and simulation demonstrated several productive interactions of durlobactam in the active sites of BlaC, PonA1, and LdtMt2. Antibiotic susceptibility testing was conducted on 11 Mtb isolates with amoxicillin, ceftriaxone, meropenem, imipenem, clavulanate, and durlobactam. Durlobactam had a minimum inhibitory concentration (MIC) range of 0.5–16 μg/mL, similar to the ranges for meropenem (1–32 μg/mL) and imipenem (0.5–64 μg/mL). In β-lactam + durlobactam combinations (1:1 mass/volume), MICs were lowered 4- to 64-fold for all isolates except one with meropenem–durlobactam. This work supports further exploration of novel β-lactamase inhibitors that target BlaC and Mtb peptidoglycan transpeptidases.
期刊介绍:
ACS Infectious Diseases will be the first journal to highlight chemistry and its role in this multidisciplinary and collaborative research area. The journal will cover a diverse array of topics including, but not limited to:
* Discovery and development of new antimicrobial agents — identified through target- or phenotypic-based approaches as well as compounds that induce synergy with antimicrobials.
* Characterization and validation of drug target or pathways — use of single target and genome-wide knockdown and knockouts, biochemical studies, structural biology, new technologies to facilitate characterization and prioritization of potential drug targets.
* Mechanism of drug resistance — fundamental research that advances our understanding of resistance; strategies to prevent resistance.
* Mechanisms of action — use of genetic, metabolomic, and activity- and affinity-based protein profiling to elucidate the mechanism of action of clinical and experimental antimicrobial agents.
* Host-pathogen interactions — tools for studying host-pathogen interactions, cellular biochemistry of hosts and pathogens, and molecular interactions of pathogens with host microbiota.
* Small molecule vaccine adjuvants for infectious disease.
* Viral and bacterial biochemistry and molecular biology.