title: Structural requirements for the stability of novel cephalosporins to AmpC beta-lactamase based on 3D-structure.
abstract: AmpC beta-lactamase is one of the leading causes of Pseudomonas aeruginosa (P. aeruginosa) resistance to cephalosporins. FR259647 is a cephalosporin having a novel pyrazolium substituent at the 3-position and exhibits excellent activity (MIC=1 microg/mL) against the AmpC beta-lactamase overproducing P. aeruginosa FP1380 strain in comparison with the third-generation cephalosporins FK518 [Abstracts of Papers, 30th Interscience Conference on Antimicrobial Agents and Chemotherapy, Atlanta, GA, October 21-24, 1990, Abs. 454; Abstracts of Papers, 30th Interscience Conference on Antimicrobial Agents and Chemotherapy, Atlanta, GA, October 21-24, 1990, Abs. 455; Abstracts of Papers, 30th Interscience Conference on Antimicrobial Agents and Chemotherapy, Atlanta, GA, October 21-24, 1990, Abs. 456; Abstracts of Papers, 30th Interscience Conference on Antimicrobial Agents and Chemotherapy, Atlanta, GA, October 21-24, 1990, Abs. 457] (MIC=16 microg/mL) and ceftazidime (CAZ) (MIC=128 microg/mL). The stability of FR259647 and FK518 to AmpC beta-lactamase was evaluated using MIC assays against both the P. aeruginosa PAO1 strain and a PAO1 mutant strain overproducing AmpC beta-lactamase as a differential assay, which indicates that the main difference derives from their stability to AmpC beta-lactamase. A structural analysis using computer simulations indicated that the difference in stability may be due to steric hindrance of the 3-position substituents causing differential affinity. This steric hindrance may disturb entry of the cephalosporins into the binding pocket. We predicted the possibility of inhibition of entry as a potential means of enhancing stability by conformational analysis. In order to validate this speculation, novel FR259647 derivatives 4-9 were designed, calculated, synthesized, and evaluated. As a result, we demonstrated that their probability of entry correlated with the MIC ratio of the mutant strain to the parent strain and supports the validity of our model.
authors: Lead Discovery Research Labs, Astellas Pharma Inc., 21 Miyukigaoka, Tsukuba, Ibaraki 305-8585, Japan.
authors: Kenji
authors: K
authors: Murano
authors: Toshio
authors: T
authors: Yamanaka
authors: Ayako
authors: A
authors: Toda
authors: Hidenori
authors: H
authors: Ohki
authors: Shinya
authors: S
authors: Okuda
authors: Kohji
authors: K
authors: Kawabata
authors: Kazuo
authors: K
authors: Hatano
authors: Shinobu
authors: S
authors: Takeda
authors: Hisashi
authors: H
authors: Akamatsu
authors: Kenji
authors: K
authors: Itoh
authors: Keiji
authors: K
authors: Misumi
authors: Satoshi
authors: S
authors: Inoue
authors: Tatsuya
authors: T
authors: Takagi
mesh_terms: D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D002511:Cephalosporins / Q000737:chemistry* / Q000494:pharmacology*; D004795:Enzyme Stability / Q000187:drug effects; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D015394:Molecular Structure; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D018440:beta-Lactam Resistance / Q000187:drug effects; D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002511:Cephalosporins; D065093:beta-Lactamase Inhibitors; C048795:AmpC beta-lactamases; D001618:beta-Lactamases