Affinity of cefditoren for penicillin-binding proteins in bacteria and its relationship with antibiotic sensitivity.

Penicillin-binding proteins (PBPs) are the targets of β-lactam antibiotics; however, changes in the affinity of PBPs for beta-lactam antibiotics often affect the susceptibility of bacteria to antibiotics. The purpose of this study was to elucidate the mechanism by which cefditoren, an oral third-generation cephalosporin, binds PBPs. The minimal inhibitory concentration (MIC), bactericidal curves, and inhibition zone comparisons were assessed to evaluate the antibacterial activity of cefditoren. PBP1A and PBP2X proteins from Streptococcus pneumoniae were purified, and their ability to bind to cefditoren was investigated via microscale thermophoresis. The Kd of cefditoren toward PBP1A was 0.005 ± 0.004 µM, which was lower than those of other cephalosporins (cefcapene, cefixime and cefdinir). In contrast, the Kd of cefditoren toward PBP2X of S. pneumoniae was 9.70 ± 8.24 µM, which was lower than that of cefixime but higher than those of cefcapene and cefdinir. Additionally, the biotinylated ampicillin (BIO-AMP) method was employed to evaluate the affinity of cefditoren toward PBPs of Haemophilus influenzae, and the results demonstrated that cefditoren and PBP3A/B had the lowest IC