Doripenem in vitro Doripenem in vitro 7 7 3 DRPM in vitro DRPM 00 DRPM Staphylococcus Streptococcus µ gml MIC90 Moraxella catarrhalishaemophilus influenzae µ gml MIC90 imipenem IPM panipenem meropenemmepm Pseudomonas aeruginosa MIC50MIC90 µ g ml P. aeruginosa DRPM ph Staphylococcus aureus Escherichia coli MBC time-kill study DRPM metallo-β -lactamase β -lactamase class ACD β -lactamase DRPM penicillin-binding proteinpbpdrpm MEPM S. aureus PBPP. aeruginosa PBPPBP3E. coli PBP S. aureusp. aeruginosa E. coli DRPM MEPM IPM MIC Key words: antibacterial activitypenicillin-binding protein DoripenemDRPM 3 Fig. imipenem IPM ceftazidimecazpseudomonas aeruginosa β -lactamase DRPM penicillinbinding proteinpbp I DRPMimipenemIPMU. S. PharmacopeiameropenemMEPMpanipenem 3 HO H 3 C H O Fig.. CO H N 3 H H H H H NH CH 3 S N H Chemical structure of. O S O H O NH PAPMbiapenemBIPM ceftazidimecazu. S. Pharmacopeiapenicillin GPCG U. S. PharmacopeiaoxacillinMPIPCU. S. PharmacopeiaampicillinABPCU. S. PharmacopeiavancomycinamikacinU. S. PharmacopeiasulbactamcefoperazoneSBTCPZU. S. PharmacopeiacephalothinCET SBTCPZ CPZ
Table.Antibacterial spectrum of Organism meropenem MIC g/ml imipenem panipenem biapenem ceftazidime Aerobic bacteria Staphylococcus aureus Staphylococcus epidermidis Staphylococcus haemolyticus Staphylococcus saprophyticus Streptococcus pyogenes Streptococcus agalactiae Streptococcus pneumoniae Streptococcus mitis Streptococcus oralis Streptococcus sanguis Streptococcus parasanguis Streptococcus gordonii Streptococcus crista Streptococcus anginosus Enterococcus faecalis Enterococcus faecium Enterococcus avium Micrococcus luteus Corynebacterium diphtheriae Bacillus anthracis Bacillus cereus Bacillus subtilis Escherichia coli Shigella dysenteriae Shigella flexneri Shigella sonnei Salmonella typhi Salmonella paratyphi Salmonella enteritidis Citrobacter freundii Klebsiella pneumoniae Klebsiella oxytoca Enterobacter cloacae Enterobacter aerogenes Serratia marcescens Proteus mirabilis Proteus vulgaris Providencia rettgeri Providencia stuartii Providencia alcalifaciens Morganella morganii Yersinia enterocolitica Yersinia pseudotuberculosis Vibrio fluvialis Vibrio vulnificus FDA 09P JC- 0.00 0.00 ATCC593 SR a SR3 b 3 3 SR33 b ATCC990 SR97 c 3 3 ATCC9970 0.00 0.00 SR d ATCC5305 ATCC039 0.00 0.00 0.00 0.00 0.00 ATCC995 0.00 0.00 TYPE III 0.00 0.00 0.00 0.00 0.00 ATCC99 e SR99 f ATCC95 ATCC9 ATCC055 ATCC59 0.00 0.00 0.00 0.00 ATCC055 0.00 ATCC500 ATCC33397 0.00 ATCC9 NCTC77 ATCC05 ATCC93 TORONT 0.00 0.00 ATCC57 IFO300 3 ATCC33 NIHJ JC- ATCC59 SR007 SR00 ATCC00 90 05 G- ATCC090 ATCC33 ATCC3 ATCC307 ATCC30 ATCC30 ATCC990 ATCC335 ATCC99 ATCC99 ATCC9 ATCC530 ATCC90 ATCC933 NCTC37 ATCC75 Continue
Doripenem in vitro Organism Aeromonas hydrophila Haemophilus influenzae Haemophilus parainfluenzae Moraxella catarrhalis Neisseria gonorrhoeae Neisseria meningitidis Pseudomonas aeruginosa Pseudomonas putida Burkholderia cepacia Stenotrophomonas maltophilia Acinetobacter baumannii Anaerobic bacteria Peptostreptococcus anaerobius Peptostreptococcus asaccharolyticus Peptostreptococcus magnus Peptostreptococcus micros Peptostreptococcus prevotii Staphylococcus saccharolyticus Propionibacterium acnes Bifidobacterium adolescentis Bifidobacterium bifidum Bifidobacterium longum Eubacterium aerofaciens Eubacterium limosum Lactobacillus acidophilus Lactobacillus casei Lactobacillus delbrueckii Lactobacillus ruminis Clostridium difficile Clostridium perfringens Clostridium sporogenes Mobiluncus curtisii Mobiluncus holmesii Mobiluncus mulieris Bacteroides fragilis Bacteroides thetaiotaomicron Bacteroides vulgatus Bacteroides distasonis Bacteroides ovatus Bacteroides uniformis Prevotella bivia Prevotella melaninogenica Prevotella intermedia Porphyromonas asaccharolytica Fusobacterium mortiferum Fusobacterium necrophorum Fusobacterium nucleatum Fusobacterium varium Veillonella parvula Capnocytophaga ochracea meropenem MIC g/ml imipenem panipenem biapenem ceftazidime IFO30 ATCC97 ATCC97 ATCC0 g ATCC33533 h ATCC790 ATCC53 0.00 0.00 0.00 0.00 0.00 ATCC37 i 0.00 ATCC9 IID5 ATCC753 ATCC59 SR05 j ATCC33 ATCC5 3 ATCC337 3 ATCC90 ATCC7337 ATCC93 ATCC93 ATCC3370 ATCC93 ATCC953 ATCC7 JCM50 JCM ATCC5707 ATCC59 ATCC IFO305 IFO3533 ATCC797 ATCC770 ATCC757 ATCC3 ATCC35 ATCC35 ATCC35 ATCC353 ATCC55 ATCC97 ATCC937 ATCC503 ATCC3 ATCC9 ATCC9303 GAI590 ATCC5 ATCC50 ATCC97 ATCC5 ATCC55 ATCC50 ATCC0790 ATCC3359 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 MICs were determined by broth microdilution or agar dilution methods as recommended by NCCLS. a penicillinase-producing strain, b MRSA, c MRSE, d methicillin, teicoplanin-resistant strain, e penicillin-intermediate strain, f penicillin-resistant strain, g serotype-b, h serotype-b, -lactamase-producing strain, i -lactamase-producing strain, j metallo- -lactamase-producing strain 3 3 3 3
Doripenem in vitro
Doripenem in vitro 5 00 3MIC National Committee for Clinical Laboratory Standards NCCLS 3, broth microdilution Neisseria gonorrhoeae agar dilution MIC cation Mueller-Hinton brothcamhbbecton Dickinson 5 Wilkins-Chalgren agarbecton DickinsonStreptococcus Haemophilus yeast extract0.005 NAD 5 CAMHB N. gonorrhoeae supplement GC Becton Dickinson Staphylococcus MPIPC MIC NaCl CAMHB 3 MIC CAMHB
Table 3. Effects of factors on MIC g/mlof Broth medium Inoculum sizecfu/ml f Medium ph f Inactivated horse serum% f CAMHB a.5 0 BHI b HIB c TSB d NB e 5 0 5 0 5 5 0 5.5 7.0 0 50 0.00 0.00 0.00 0.00 0.00 0.00 Organism Staphylococcus aureus FDA 09P JC- ATCC593 Staphylococcus epidermidis ATCC990 Enterococcus faecalis ATCC933 ATCC9 Escherichia coli NIHJ JC- ATCC59 Citrobacter freundii ATCC090 Klebsiella pneumoniae ATCC33 Enterobacter cloacae ATCC307 Serratia marcescens ATCC30 Pseudomonas aeruginosa ATCC753 ATCC59 a Cation-adjusted Mueller Hinton broth, b Brain heart infusion broth, c Heart infusion broth, d Tryptic soy broth, e Nutrient broth, f MICs were determined by a broth microdilution method with CAMHB. brain heart infusion brothheart infusion brothtryptic soy brothnutrient broth Becton Dickinson 50 5 CFUmL 50 50 CFUmL ph NaOH HCl ph 5.5 7.0.5 CAMHB 0 50 CAMHB 5MBC Staphylococcus aureus Escherichia coli 00 NCCLS macrodilution 5 CAMHB 35 brain heart infusion agarbecton Dickinson 37,000 MBC Time-kill study S. aureus Smith E. coli NIHJ JC- 0 CFUmL CAMHB 37.5 7PBP S. aureus SmithP. aeruginosa ATCC59E. coli NIHJ JC- CPCG Spratt SDS PBP BAS000 CPCG PBP 50 IC50 β -Lactamase β - lactamase 7 β -lactamase 30 β β -lactamase class ACD00 µ moll ABPC CET Metallo-β -lactamase Km CET VmaxMichaelis-Menten 9 S. aureus SmithP. aeruginosa ATCC59E. coli NIHJ JC- CAMHB 35 3 5 MIC NCCLS broth microdi-
Doripenem in vitro Table. Bactericidal activity of against clinical isolates of Staphylococcus aureus and Escherichia coli OrganismNo. of strains Staphylococcus aureus -lactamase-producing strain0 -lactamase-nonproducing strain0 Escherichia coli0.30.30.90 Mean of MBC/MIC ratios a of meropenem.90.0.0 imipenem.30.0.05 ceftazidime a MICs and MBCs were determined by a broth macrodilution method as recommended by NCCLS, and the mean of MBC/MIC ratios for 0 or 0 strains was calculated..0.0.0 MIC: g/ml meropenem MIC: g/ml imipenem MIC: g/ml ceftazidime MIC: g/ml Viable cellslog CFU/mL control / MIC / MIC MIC MIC MIC MIC 0 0 0 0 Incubation timeh MIC: g/ml meropenem MIC: g/ml imipenem MIC: g/ml ceftazidime MIC: g/ml Viable cellslog CFU/mL control / MIC / MIC MIC MIC MIC MIC 0 0 0 0 Incubation timeh Fig.. Killing kinetic curves of against Staphylococcus aureus SmithAand Escherichia coli NIHJ JC-B. lution II 9 09 DRPM MEPMIPMPAPM BIPMCAZ MIC Table DRPM 0.00 µ gml MIC Streptococcus pneumoniae PRSP Haemophilus influenzae β -lactamase DRPM Staphylococcus P. aeruginosa metalloβ -lactamase Stenotrophomonas maltophilia DRPM 0.00 µ gml MIC DRPM Table
Table 5. Binding affinity of to penicillin-binding proteins Sorce of bacterial membrane PBP IC50 g/ml a meropenem imipenem 0.07 0.09 0.0 Staphylococcus aureus Smith 3.5 3 0.0 0.0 0. 0.0 0.0 A 0.0 0. 0.9 B 0.7 0. 0.75 0.3 0.09 0.9 Pseudomonas aeruginosa ATCC59 3 0.0 0.055 0.7 0.0 5.9. A 0. 0. B. 0.3 0.90 0.055 Escherichia coli NIHJ JC- 3. 0.5. 0.05 5 5 0. 0.9 a Binding affinity was expressed as the concentrationic50at which binding of Cbenzylpenicillin toward each PBP was inhibited by 50% of that in the control without the addition of antibiotics. DRPM Staphylococcus DRPM MIC90 3 µ gml DRPM S. aureus Staphylococcus epidermidis coagulase Staphylococcus µ gml MIC90 DRPM Streptococcus S. pneumoniae µ gml MIC90 S. pneumoniae PRSP µ gml MIC90 Streptococcus pyogenesstreptococcus agalactiaestreptococcus anginosus group µ gml MIC90 Enterococcus Enterococcus faecalis DRPM MIC50 MIC90 µ gml Enterococcus faecium DRPM MIC90 DRPM IPM PAPM MEPM BIPM Moraxella catarrhalis DRPM µ gml MIC90 DRPM IPMPAPM BIPMCAZ MEPM DRPM H. influenzae β -lactamase ABPC BLNAR MIC90 µ g ml β-lactamase 3 BLNAR MIC90 µ gml H. influenzae DRPM MEPMCAZ MIC P. aeruginosa DRPM MIC50 µ gmlmic90 µ gml Acinetobacter DRPM µ gml MIC90 Burkholderia cepacia MIC50MIC90 µ gml Peptostreptococcus Prevotella DRPM µ gml µ g ml MIC90 Clostridium difficile Bacteroides DRPM MIC90 µ gml 3 MIC 3 CAMHB nutrient broth MIC 5 MIC
Doripenem in vitro Table 3 MIC 50 5 CFUmL 50 CFU ml MIC 50 CFUmL MIC 3 -lactamases Table. Stability of against hydrolysis by class A, C, and D Relative hydrolysis rate a MIC g/ml b for source strain meropenem imipenem cephalothin ampicillin meropenem imipenem Source of enzyme class S. aureus SR5 A 0. 0. 0. f 00 K. pneumoniae GN9 A 0. 0. 0. 00 E. coli W30/RTEM c A 0. 0. 0. 00 P. vulgaris SR3 A 0.00 0.00 0.00 00 E. coli SHMR/pKP95 AESBL e 0.0 0.0 0.0 00 E. cloacae SR3 C 0.00 0.00 0.00 00 C. freundii SR9 C 0.0 0.0 0.0 00 P. aeruginosa SR- C 0.00 0.00 0.00 00 E. coli ML0/RGN3 d D 0.7 0.7 0.7 00 a Expressed relative to arbitrary 00 for cephalothin or ampicillin at a concentration of 00 mm, b MICs were determined by a broth microdilution method as recommended by NCCLS, c TEM- type, d OXA- type, e extended spectrum -lactamasetoho--like type, f not tested ph MIC S. aureus E. faecalis MIC MIC E. coli S. aureus Citrobacter freundii 50 MIC MIC β -lactamase S. aureus E. coli 00 0 MBC Table MBCMIC S. aureus β -lactamase DRPM MBC MIC E. coli DRPM DRPMMEPMCAZ E. coli MBCMIC IPM 3 S. aureus Smith E. coli NIHJ JC- DRPM Fig. DRPM DRPM MIC,000 5PBP S. aureus SmithP. aeruginosa ATCC59E. coli NIHJ JC- PBP DRPM Table 5 S. aureus PBP DRPM PBP PBP P. aeruginosa DRPM PBP3 PBPAB E. coli DRPM PBP PBPB3 S. aureus P. aeruginosa PBP DRPM MEPM β -Lactamase DRPM β -lactamaseextended-spectrum β -lactamaseesbl class ACD β -lactamase Table DRPM MEPM IPM P. aeruginosa class C β -lactamase DRPM relative VmaxKm metallo-β -lactamase Table 7
Source of enzyme Table 7. Stability of against hydrolysis by metallo- -lactamases Parameter meropenem imipenem cephalothin Km mol/l 35 3.7.5 Escherichia coli SHMR/pKP9 a Relative Vmax b 0 3 0 00 Relative Vmax/Km c.3 5 0 00 MIC g/ml d e Km mol/l. 7.5 Stenotrophomonas maltophilia SR7 Relative Vmax 9 0 00 Relative Vmax/Km 7 9 59 00 MIC g/ml a IMP- type, b Expressed relative to arbitrary 00 for Vmax of cephalothin. c Expressed relative to arbitrary 00 for Vmax/Km of cephalothin. d MICs were determined by a broth microdilution method as recommended by NCCLS, e not tested A B C g/ml g/ml g/ml Fig. 3. Acquired resistance of Staphylococcus aureus SmithA, Escherichia coli NIHJ JC-B,andPseudomonas aeruginosa ATCC59Cwhen subcultured repeatedly in media containing, meropenem, or imipenem.
Doripenem in vitro 7 S. aureus SmithE. coli NIHJ JC-P. aeruginosa ATCC59 DRPM MEPMIPM 3 Fig. 3S. aureus IPM DRPM MEPM E. coli DRPM IPM MEPM P. aeruginosa MIC 3 III DRPM Table DRPM IPM PAPM MEPM P. aeruginosa IPM MEPM DRPM Tsuji Nomura 9 Ge 0 Jones DRPM ph MBC timekill study IPM MEPM DRPM metalloβ -lactamase class ACD β -lactamase Toho- ESBL Mushtaq AmpC ESBL DRPM IPM MEPM AmpC β -lactamase DRPM Table MIC90 µ gml β -lactamase PBP DRPM S. aureus PBP PBP P. aeruginosa PBP PBP3 E. coli PBP DRPM PBP DRPM IC50 MEPM DRPM in vitro DRPM in vivo,3 dehydropeptidase-i I MEPM 5 Iso Y, Irie T, Nishino Y, et al: A novel β -methylcarbapenem antibiotic, S-. Synthesis and structure-activity relationships of -5-substituted pyrrolidin-3-ylthio-β -methylcarbapenems. J Antibiot 9: 9909, 99 Tsuji M, Ishii Y, Ohno A, et al: In vitro and in vivo antibacterial activities of S-, a new carbapenem. Antimicrob Agents Chemother : 999, 99 3 National Committee for Clinical Laboratory Standards: Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard-fifth edition, M7-A5. National Committee for Clinical Laboratory Standards, Wayne, Pa, 000 National Committee for Clinical Laboratory Standards: Methods for antimicrobial susceptibility testing of anaerobic bacteria; approved standard-fifth edition, M-A5. National Committee for Clinical Laboratory Standards, Wayne, Pa, 00 5 National Committee for Clinical Laboratory Standards: Methods for determining bactericidal activity of antimicrobial agents; approved guideline, M-A. National Committee for Clinical Laboratory Standards, Wayne, Pa, 999 Spratt B G: Properties of the penicillin-binding proteins of Escherichia coli K. Eur J Biochem 7: 335, 977 7 Murakami K, Yoshida T: Covalent binding of moxalac-
tam to cephalosporinase of Citrobacter freundii. Antimicrob Agents Chemother 7: 7773, 95 Jasson J A T: A direct spectrometric assay for penicillin β -lactamasepenicillinase. Biochim Biophys Acta 99: 77, 95 9 Nomura S, Nagayama A: In vitro antibacterial activity of S-, a new parenteral carbapenem, against urological pathogens isolated from patients with complicated urinary tract infections. J Chemother : 55 0, 00 0 Ge Y, Wikler M A, Sahm D F, et al: In vitro antimicrobial activity of, a new carbapenem. Antimicrob Agents Chemother : 339, 00 Jones R N, Huynh H K, Biedenbach D J, et al: DoripenemS-, a novel carbapenem: comparative activity against contemporary pathogens including bacterial action and preliminary in vitro methods evaluation. J Antimicrob Chemother 5: 5, 00 Mushtaq S, Ge Y, Livermore D M: Comparative activities of versus isolates, mutants, and transconjugants of Enterobacteriaceae and Acinetobacter spp. with characterized β -lactamases. Antimicrob Agents Chemother : 3339, 00 3 Doripenem in vivo 53Suppl :779, 005 Doripenem dehydropeptidase-i 53 Suppl :995, 005 5 Doripenem I 53Suppl : 03, 005 In vitro antibacterial activity of, a novel parenteral carbapenem Takaji Fujimura, Yoshiji Kimura, Isamu Yoshida, Isao Higashiyama, Yutaka Jinushi, Tadashi Munekage, Naomi Kuroda, Masayoshi Doi, Toru Nishikawa and Yoshinori Yamano Discovery Research Laboratories, Shionogi & Co., Ltd., 3 Futaba-cho, Toyonaka, Osaka, Japan We studied the in vitro antibacterial activity of DRPM, a novel parenteral carbapenem. DRPM showed a broad antibacterial spectrum against aerobic gram-positive and negative bacteria and anaerobic bacteria. In a susceptibility test for clinical strains isolated in 00, MIC90s ofdrpmwere- µ gml for Streptococcus spp. and methicillin-susceptible strains of Staphylococcus spp., and - µ gml for Enterobacteriaceae, Moraxella catarrhalis, andhaemophilus influenzae, suggesting that DRPM has potent antibacterial activity. Of the 5 carbapenems tested, DRPM was most potent against aerobic gram-positive bacteria, following imipenemipmand panipenem, and is most potent against aerobic gram-negative bacteria, following meropenemmepm. MIC50 and MIC90 of DRPM against Pseudomonas aeruginosa were and µ gml, indicating that DRPM has the most potent antipseudomonal activity among antibiotics tested. The antibacterial activity of DRPM was not affected by factors such as medium ph or inoculum size. Results from MBC determination and a time-kill study suggested that DRPM has strong bactericidal activity corresponding to its antibacterial activity. Although DRPM was hydrolyzed by metallo-β -lactamases as well as other carbapenems, it was stabletohydrolysisbyclassa,c,anddβ -lactamases, including extended-spectrum β -lactamase. A study on the affinity of DRPM to penicillin-binding proteins as its mode of action suggested that DRPM probably exhibits antibacterial activity by inhibiting the activity of PBP for S. aureus, PBP and PBP3 for P. aeruginosa, andpbp for E. coli as well as MEPM. When subcultured repeatedly in media containing DRPM, MEPM, or IPM, bacterial strains of S. aureus, P. aeruginosa, and E. coli showed decreased susceptibility to all of these antibiotics, and cross-resistance among tested carbapenems was observed.