Inhibition of β-lactamases of mycobacteria by avibactam and clavulanate

Soroka, Daria; Ourghanlian, Clément; Compain, Fabrice; Fichini, Marion; Dubée, Vincent; Mainardi, Jean–Luc; Hugonnet, Jean-Emmanuel; Arthur, Michel

doi:10.1093/jac/dkw546

Cited by 41 publications

(50 citation statements)

References 22 publications

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“…This effect was compensated for to a limited extent by a moderate reduction (20-fold) of the decarbamylation rate. Thus, the N 132 G substitution decreased the efficacy of inhibition of both KPC-2 and CTX-M-15, as previously shown for class A ␤-lactamases from mycobacteria (9).…”

Section: Resultssupporting

confidence: 80%

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Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N ¹³² G in the Conserved SDN Motif

Ourghanlian

Soroka

Arthur

2017

Antimicrob Agents Chemother

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The substitution N 132 G in the SDN motif of class A ␤-lactamases from rapidly growing mycobacteria was previously shown to impair their inhibition by avibactam but to improve the stability of acyl-enzymes formed with clavulanate. The same substitution was introduced in KPC-2 and CTX-M-15 to assess its impact on ␤-lactamases from Enterobacteriaceae and evaluate whether it may lead to resistance to the ceftazidime-avibactam combination. Kinetic parameters for the inhibition of the ␤-lactamases by avibactam and clavulanate were determined by spectrophotometry using nitrocefin as the substrate. The substitution N 132 G impaired (Ͼ1,000-fold) the efficacy of carbamylation of KPC-2 and CTX-M-15 by avibactam. The substitution improved the inhibition of KPC-2 by clavulanate due to reduced deacylation, whereas the presence or absence of N 132 G resulted in the inhibition of CTX-M-15 by clavulanate. The hydrolysis of amoxicillin and nitrocefin by KPC-2 and CTX-M-15 was moderately affected by the substitution N 132 G, but that of ceftazidime, ceftaroline, and aztreonam was drastically reduced. Isogenic strains producing KPC-2 and CTX-M-15 were constructed to assess the impact of the substitution N 132 G on the antibacterial activities of ␤-lactam-inhibitor combinations. For amoxicillin, the substitution resulted in resistance and susceptibility for avibactam and clavulanate, respectively. For ceftazidime, ceftaroline, and aztreonam, the negative impact of the substitution on ␤-lactamase activity prevented resistance to the ␤-lactam-avibactam combinations. In conclusion, the N 132 G substitution has profound effects on the substrate and inhibition profiles of class A ␤-lactamases, which are largely conserved in distantly related enzymes. Fortunately, the substitution does not lead to resistance to the ceftazidime-avibactam combination. KEYWORDS ␤-lactamase inhibitor, avibactam, CTX-M-15, clavulanate, KPC-2 Avibactam is the first representative of a new family of inhibitors active against ␤-lactamases of classes A and C and certain enzymes of class D (1). Like ␤-lactamcontaining inhibitors of the first generation, such as clavulanate, sulbactam, and tazobactam, avibactam acts as a suicide substrate and forms a covalent adduct with the active-site serine of the enzymes (Fig. 1). However, the carbamylation reaction is reversible in the case of avibactam, whereas the acylation reaction is irreversible in the case of first-generation inhibitors (2). This difference may be accounted for by the presence of a five-membered ring in avibactam, which is sterically less constrained than the four-membered ring of ␤-lactams. Consequently, the efficacy of the inhibition of the ␤-lactamases by avibactam depends upon equilibrium between the native (active) and carbamylated (inactive) forms of the enzyme (Fig. 1). This is evaluated by determining the kinetic parameters for the on (k 2 /K i ) and off (k off ) reactions using nitrocefin

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Section: Resultssupporting

confidence: 80%

“…The substitution N 132 G also drastically reduced the efficacy of clavulanate hydrolysis by KPC-2 (Table 1). Both effects of N 132 G appear to be conserved in class A ␤-lactamases, since they have also been detected for Bla Mab from M. abscessus (9).…”

Section: Discussionmentioning

confidence: 76%

Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N ¹³² G in the Conserved SDN Motif

Ourghanlian

Soroka

Arthur

2017

Antimicrob Agents Chemother

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“…Carbapenems and classical ␤-lactamase inhibitors may provide alternatives for use against variants with impaired inhibition by avibactam, as previously observed for substitutions in the ␤-lactamases from mycobacteria (21). These observations indicate, as previously discussed (21), that the formulation of ␤-lactamase inhibitors independently from a ␤-lactam partner would provide clinicians with access to potentially useful therapeutic regimens based on combining ␤-lactams with approved inhibitors.…”

supporting

confidence: 55%

“…Recently, Shields et al also reported such reversions, but resistance to carbapenems and ceftazidime-avibactam has been obtained by multistep exposure (Ն4 weeks) of ceftazidime-avibactamresistant K. pneumoniae strains to meropenem, mainly due to modifications of the porin OmpK36 (20). Carbapenems and classical ␤-lactamase inhibitors may provide alternatives for use against variants with impaired inhibition by avibactam, as previously observed for substitutions in the ␤-lactamases from mycobacteria (21). These observations indicate, as previously discussed (21), that the formulation of ␤-lactamase inhibitors independently from a ␤-lactam partner would provide clinicians with access to potentially useful therapeutic regimens based on combining ␤-lactams with approved inhibitors.…”

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confidence: 80%

Impaired Inhibition by Avibactam and Resistance to the Ceftazidime-Avibactam Combination Due to the D ¹⁷⁹ Y Substitution in the KPC-2 β-Lactamase

Compain

Arthur

2017

Antimicrob Agents Chemother

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The ceftazidime-avibactam antibiotic combination was recently shown to be at risk for the emergence of resistance under treatment. To gain insight into the underlying mechanism, we have analyzed the catalytic properties of a Klebsiella pneumoniae carbapenemase type 2 (KPC-2) ␤-lactamase harboring the D 179 Y substitution. We show that impaired inhibition by avibactam combined with significant residual activity for ceftazidime hydrolysis accounts for the resistance. In contrast, the D 179 Y substitution abolished the hydrolysis of aztreonam and imipenem, indicating that these drugs might provide therapeutic alternatives.KEYWORDS ␤-lactamase inhibitor, avibactam, KPC-2, carbapenemase, ceftazidime R esistance to carbapenems in Enterobacteriaceae is often due to the production of class A ␤-lactamases belonging to the Klebsiella pneumoniae carbapenemase (KPC) type (1). The most commonly encountered variants worldwide are KPC-2 and KPC-3, which confer high-level resistance to most available ␤-lactams. The production of KPC ␤-lactamases is often associated with resistance to other classes of antibiotics, including aminoglycosides, fluoroquinolones, and colistin, leaving few or no therapeutic alternatives (2, 3). Inhibition of KPC enzymes by classical ␤-lactamase inhibitors (clavulanate, sulbactam, and tazobactam) is not sufficient to restore the activity of ␤-lactams (4). In this context, a new ␤-lactam-␤-lactamase inhibitor combination, ceftazidime-avibactam (5), has recently obtained regulatory approval in the United States and Europe. The combination has a broad spectrum of activity against multidrug-resistant Enterobacteriaceae and Pseudomonas aeruginosa strains producing class A, C, and some class D ␤-lactamases (6, 7).The emergence of resistance to the ceftazidime-avibactam combination has been recently reported in three out of 37 patients infected with carbapenem-resistant Enterobacteriaceae and treated with these drugs (8). The emergence of resistance, which was associated with microbiological failure, was due to amino acid substitutions in the KPC-3 enzymes produced by the K. pneumoniae isolates recovered from these three patients (D 179 Y, V 240 G, and D 179 Y associated with T 243 M) (9). The impact of these substitutions on the kinetic parameters for hydrolysis of ceftazidime and inhibition by avibactam has not been previously reported. In another study (10), site-directed mutagenesis identified rare substitutions resulting in resistance to the ceftazidimeavibactam combination. A comparison of KPC-2 and KPC-2 D 179 N did not reveal any modification of the steady-state rate of ceftazidime hydrolysis or inhibition by avibactam (10). However, an initial burst of ceftazidime hydrolysis was detected for KPC-2 D 179 N but not for the parental enzyme (10).To gain insight into the modifications of the catalytic properties of KPC enzymes that

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“…Here, we have studied if avibactam, a recently developed β-lactamase inhibitor, can alter the potency of the carbapenem class of β-lactams against M. abscessus. Recent studies have provided insight into the activities of some older carbapenems with or without avibactam against M. abscessus [16][17][18]. We have included all commercially available carbapenems, most importantly new and oral carbapenems, and a collection of clinically isolated M. abscessus strains most of which are resistant to multiple drugs currently deployed to treat infection by this pathogen.…”

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confidence: 99%

Combinations of Avibactam and Carbapenems Exhibit Enhanced Potencies Against Drug-Resistant Mycobacterium Abscessus

et al. 2017

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Aim:The objective of this study was to assess if avibactam, a new β-lactamase inhibitor, can restore the potency of carbapenems, a sub-class of β-lactams, against Mycobacterium abscessus clinical isolates. Materials & methods: 28 M. abscessus clinical isolates that are resistant to multiple drugs currently used to treat its infection were included. MIC of carbapenems alone and in combination with avibactam against these strains were determined. Results: Tebipenem, an oral carbapenem, and ertapenem and panipenem exhibited the greatest shift in MIC when supplemented with avibactam. Conclusion: Avibactam restores MICs of tebipenem, ertapenem and panipenem against M. abscessus to therapeutically achievable concentrations and raises the possibility of usefulness of these carbapenems to treat drug-resistant M. abscessus infections. Mycobacterium abscessus is a rapidly growing nontuberculous mycobacterium found widely in soil and water and can cause a spectrum of infections [1]. Prevalence of M. abscessus infections in the lungs of people with chronic conditions, such as cystic fibrosis is significant and can often lead to serious morbidity [2]. A survey revealed that M. abscessus is present in the sputum of approximately 13% of cystic fibrosis patients in the USA [3]. Among nontuberculous mycobacterium lung infections, M. abscessus is one of the prevalent species and often leads to a chronic and incurable disease [4][5][6]. Drug resistance in M. abscessus is steadily rising globally, making it increasingly difficult to manage infections with these strains [7]. Therefore, new drugs and novel regimens are acutely needed to treat infections with M. abscessus. An ideal new drug would inhibit a novel target so that it can be effective against M. abscessus strains that are resistant to currently used drugs.The peptidoglycan is an Achilles' heel of bacteria as agents that inhibit its biosynthesis, namely β-lactams and glycopeptides, comprise some of the most widely used class of antibacterials in modern medicine. β-lactams derive their activity by preventing formation of linkage between peptide side chains by inhibiting the transpeptidases that catalyze this reaction [8]. Recently it was demon strated that majority of the linkages in the peptidoglycan layer of M. abscessus are generated by LD-transpeptidases [9] and that this class of enzyme is selectively more susceptible to the carbapenem class of β-lactams [10][11][12]. Imipenem, a carbapenem, has superior activity compared with For reprint orders, please contact: reprints@futuremedicine.com

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Inhibition of β-lactamases of mycobacteria by avibactam and clavulanate

Abstract: G 132 N and N 132 G had opposite effects on the inhibition of BlaC and Bla Mab , indicating that these substitutions might lead to acquisition of resistance to either of the β-lactamase inhibitors, but not to both of them.

Cited by 41 publications

References 22 publications

Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N ¹³² G in the Conserved SDN Motif

Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N ¹³² G in the Conserved SDN Motif

Impaired Inhibition by Avibactam and Resistance to the Ceftazidime-Avibactam Combination Due to the D ¹⁷⁹ Y Substitution in the KPC-2 β-Lactamase

Combinations of Avibactam and Carbapenems Exhibit Enhanced Potencies Against Drug-Resistant Mycobacterium Abscessus

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Inhibition of β-lactamases of mycobacteria by avibactam and clavulanate

Abstract: G 132 N and N 132 G had opposite effects on the inhibition of BlaC and Bla Mab , indicating that these substitutions might lead to acquisition of resistance to either of the β-lactamase inhibitors, but not to both of them.

Cited by 41 publications

References 22 publications

Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N 132 G in the Conserved SDN Motif

Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N 132 G in the Conserved SDN Motif

Impaired Inhibition by Avibactam and Resistance to the Ceftazidime-Avibactam Combination Due to the D 179 Y Substitution in the KPC-2 β-Lactamase

Combinations of Avibactam and Carbapenems Exhibit Enhanced Potencies Against Drug-Resistant Mycobacterium Abscessus

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Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N ¹³² G in the Conserved SDN Motif

Inhibition by Avibactam and Clavulanate of the β-Lactamases KPC-2 and CTX-M-15 Harboring the Substitution N ¹³² G in the Conserved SDN Motif

Impaired Inhibition by Avibactam and Resistance to the Ceftazidime-Avibactam Combination Due to the D ¹⁷⁹ Y Substitution in the KPC-2 β-Lactamase