In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase
β-Lactamase-mediated resistance to β-lactam antibiotics has been significantly threatening the efficacy of these clinically important antibacterial drugs. Although some β-lactamase inhibitors are prescribed in combination with β-lactam antibiotics to overcome this resistance, the emergence of enzyme...
| Published in: | Frontiers in Cellular and Infection Microbiology |
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| Main Authors: | , , , , , , |
| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-12-01
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| Online Access: | https://www.frontiersin.org/article/10.3389/fcimb.2018.00441/full |
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| author | Jung-Hyun Na Tae Hee Lee Tae Hee Lee Soo-Bong Park Min-Kyu Kim Min-Kyu Kim Bo-Gyeong Jeong Kyung Min Chung Kyung Min Chung Sun-Shin Cha |
| author_facet | Jung-Hyun Na Tae Hee Lee Tae Hee Lee Soo-Bong Park Min-Kyu Kim Min-Kyu Kim Bo-Gyeong Jeong Kyung Min Chung Kyung Min Chung Sun-Shin Cha |
| author_sort | Jung-Hyun Na |
| collection | DOAJ |
| container_title | Frontiers in Cellular and Infection Microbiology |
| description | β-Lactamase-mediated resistance to β-lactam antibiotics has been significantly threatening the efficacy of these clinically important antibacterial drugs. Although some β-lactamase inhibitors are prescribed in combination with β-lactam antibiotics to overcome this resistance, the emergence of enzymes resistant to current inhibitors necessitates the development of novel β-lactamase inhibitors. In this study, we evaluated the inhibitory effect of dinucleotides on an extended-spectrum class C β-lactamase, AmpC BER. Of the dinucleotides tested, NADPH, a cellular metabolite, decreased the nitrocefin-hydrolyzing activity of the enzyme with a Ki value of 103 μM in a non-covalent competitive manner. In addition, the dissociation constant (KD) between AmpC BER and NADPH was measured to be 40 μM. According to our in vitro susceptibility study based on growth curves, NADPH restored the antibacterial activity of ceftazidime against a ceftazidime-resistant Escherichia coli BER strain producing AmpC BER. Remarkably, a single dose of combinatory treatment with NADPH and ceftazidime conferred marked therapeutic efficacy (100% survival rate) in a mouse model infected by the E. coli BER strain although NADPH or ceftazidime alone failed to prevent the lethal bacterial infection. These results may offer the potential of the dinucleotide scaffold for the development of novel β-lactamase inhibitors. |
| format | Article |
| id | doaj-art-8a70711d5bec4d64b7d331f320b306ef |
| institution | Directory of Open Access Journals |
| issn | 2235-2988 |
| language | English |
| publishDate | 2018-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| spelling | doaj-art-8a70711d5bec4d64b7d331f320b306ef2025-08-19T21:06:46ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882018-12-01810.3389/fcimb.2018.00441429737In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-LactamaseJung-Hyun Na0Tae Hee Lee1Tae Hee Lee2Soo-Bong Park3Min-Kyu Kim4Min-Kyu Kim5Bo-Gyeong Jeong6Kyung Min Chung7Kyung Min Chung8Sun-Shin Cha9Department of Chemistry and Nanoscience, Ewha Womans University, Seoul, South KoreaDepartment of Microbiology and Immunology, Chonbuk National University Medical School, Jeonju, South KoreaInstitute for Medical Science, Chonbuk National University Medical School, Jeonju, South KoreaDepartment of Chemistry and Nanoscience, Ewha Womans University, Seoul, South KoreaBiotechnology Research Division, Korea Atomic Energy Research Institute, Jeongeup, South KoreaDepartment of Radiation Biotechnology and Applied Radioisotope Science, University of Science and Technology, Daejeon, South KoreaDepartment of Chemistry and Nanoscience, Ewha Womans University, Seoul, South KoreaDepartment of Microbiology and Immunology, Chonbuk National University Medical School, Jeonju, South KoreaInstitute for Medical Science, Chonbuk National University Medical School, Jeonju, South KoreaDepartment of Chemistry and Nanoscience, Ewha Womans University, Seoul, South Koreaβ-Lactamase-mediated resistance to β-lactam antibiotics has been significantly threatening the efficacy of these clinically important antibacterial drugs. Although some β-lactamase inhibitors are prescribed in combination with β-lactam antibiotics to overcome this resistance, the emergence of enzymes resistant to current inhibitors necessitates the development of novel β-lactamase inhibitors. In this study, we evaluated the inhibitory effect of dinucleotides on an extended-spectrum class C β-lactamase, AmpC BER. Of the dinucleotides tested, NADPH, a cellular metabolite, decreased the nitrocefin-hydrolyzing activity of the enzyme with a Ki value of 103 μM in a non-covalent competitive manner. In addition, the dissociation constant (KD) between AmpC BER and NADPH was measured to be 40 μM. According to our in vitro susceptibility study based on growth curves, NADPH restored the antibacterial activity of ceftazidime against a ceftazidime-resistant Escherichia coli BER strain producing AmpC BER. Remarkably, a single dose of combinatory treatment with NADPH and ceftazidime conferred marked therapeutic efficacy (100% survival rate) in a mouse model infected by the E. coli BER strain although NADPH or ceftazidime alone failed to prevent the lethal bacterial infection. These results may offer the potential of the dinucleotide scaffold for the development of novel β-lactamase inhibitors.https://www.frontiersin.org/article/10.3389/fcimb.2018.00441/fullantimicrobial resistanceclass C β-lactamaseAmpC BERNADPHβ-lactamase inhibitorsmouse infection model |
| spellingShingle | Jung-Hyun Na Tae Hee Lee Tae Hee Lee Soo-Bong Park Min-Kyu Kim Min-Kyu Kim Bo-Gyeong Jeong Kyung Min Chung Kyung Min Chung Sun-Shin Cha In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase antimicrobial resistance class C β-lactamase AmpC BER NADPH β-lactamase inhibitors mouse infection model |
| title | In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase |
| title_full | In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase |
| title_fullStr | In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase |
| title_full_unstemmed | In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase |
| title_short | In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase |
| title_sort | in vitro and in vivo inhibitory activity of nadph against the ampc ber class c β lactamase |
| topic | antimicrobial resistance class C β-lactamase AmpC BER NADPH β-lactamase inhibitors mouse infection model |
| url | https://www.frontiersin.org/article/10.3389/fcimb.2018.00441/full |
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