ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Feb. 1991, 0066-4804/91/020259-08$02.00/0 Copyright C) 1991, American Society for Microbiology
p.
Vol. 35, No. 2
259-266
In Vitro Activity and 3-Lactamase Stability of GR69153, a New Long-Acting Cephalosporin NAI-XUN CHIN,' JIAN-WEI GU,' WEI FANG,' AND HAROLD C. NEU' 2* Departments of Medicine' and Pharmacology,2 College of Physicians and Surgeons, Columbia University, New York, New York 10032 Received 11 September 1990/Accepted 20 November 1990
GR69153, a new parenteral cephalosporin, inhibited 90% of Escherichia coli, Klebsiella oxytoca, Proteus mirabilis, Citrobacter diversus, shigellae, and salmonellae at 32 ,ug/ml, and cefpirome and cefepime were the most active agents against these species. Pseudomonas aeruginosa was highly susceptible to GR69153, and for this organism the MIC90 was 128 0.5-4
0.06 0.12 0.25 0.03 0.03 2 1
0.25 0.25 0.5 0.12 0.12 >128 2
Klebsiella pneumoniae (30)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015->16 0.06->16 0.12->16 0.015->32 0.015->32 1->128 0.5-8
0.06 0.12 0.25 0.06 0.06 8 1
1 0.5 1 1 1 >128 4
Klebsiella oxytoca (20)
GR69153 E-1040 Ceftazidime
Cefpirome Cefepime Piperacillin Gentamicin
0.015-4 0.06-2 0.12-2 0.015-1 0.015-1 2->128 0.5-1
0.06 0.12 0.25 0.06 0.06 8 0.5
0.12 0.5 0.5 0.25 0.25 >128 1
Enterobacter aerogenes (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-32 0.06-0.5 0.12->32 0.015-1 0.015-0.5 2-128 0.25-1
0.12 0.12 0.5 0.12 0.25 4 0.5
8 0.25 32 0.5 0.5 64 1
Enterobacter cloacae (30)
GR69153 E-1040 Ceftazidime Cefpirome
Cefepime Piperacillin Gentamicin
0.015->32 0.015-4 0.015->32 0.015-16 0.015-4 32->256 0.5->16
0.25 0.12 4 0.25 0.25 128 1
>32 2 >32 16 2 >128 >16
Citrobacterfreundii (30)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015->32 0.015->32 0.015->32 0.015->32 0.015-16 1->128 0.25->16
1 0.12 1 0.12 0.06 4 1
>32 4 >32 8 8 >128 16
Citrobacter diversus (15)
GR69153 E-1040 Ceftazidime
0.015-0.06 0.015-0.12 0.015-0.25 0.015-0.06 0.015-0.06
0.06 0.12 0.12 0.06 0.06 8 0.5
0.06 0.12 0.25 0.06 0.06 32 2
0.06 0.25 0.06 0.015 0.03 0.5 1
0.06 0.5 0.12 0.03 0.06 1 16
0.5 0.06 0.06
8 0.06 0.5
0.015
0.015
Cefpirome Cefepime Piperacillin Gentamicin Proteus mirabilis (20)
GR69153 E-1040
Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
Morganella morganii (10)
GR69153 E-1040 Ceftazidime
Cefpirome
4->128 0.25-4 0.06-0.25 0.12-1 0.06-0.25 0.015-0.5 0.03-0.5 0.25-1 0.5->16
0.06-8 0.06-0.25 0.015-1 0.015-0.03
Continued on following page
ACTIVITY AND STABILITY OF GR69153
VOL. 35, 1991
261
TABLE 1-Continued Organism (no. of isolates tested)
MIC Antimicrobial (gng/ml)a agent Range 50%
90%
0.015 8 1
Cefepime Piperacillin Gentamicin
0.015-0.03 0.12-32 0.5-1
0.015 2 0.5
Proteus vulgaris (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-0.12 0.12-1 0.06-0.25 0.03-1 0.015-0.12 0.25->128 0.12->16
0.06 0.12 0.06 0.12 0.03 8 0.5
0.12 0.5 0.12 0.5 0.06 >128 2
Proteus rettgeri (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-2 0.06-0.5 0.06-1 0.06-0.5 0.06-0.5 0.25->128 1->16
2 0.25 0.25 0.25 0.25 0.5 2
2 0.5 0.5 0.5 0.5 >128 16
Providencia stuartii (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-4 0.06-0.5 0.06-1 0.06-0.25 0.06-4 0.25->128 2->16
0.06 0.06 0.12 0.06 0.06 8 8
Serratia marcescens (30)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-4 0.12-8 0.12-16 0.06-4 0.06-4 1->128 1->16
0.25 0.25 0.25 0.12 0.12 16 2
1 4 1 0.5 1 >128 8
Pseudomonas aeruginosa (45)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-8 0.25-16 1->32 2->32 1-16 2->128 2->16
0.5 0.5 2 4 2 32 4
2 1 32 32 8 >128 >16
Pseudomonas cepacia (17)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.25->64 0.12-64 2->64 0.12->64 0.12-64 2->128 0.25->16
16 32 16 8 8 8 2
>64 64 >64 >64 64 >128 >32
Xanthomonas maltophilia (12)
GR69153 E-1040 Ceftazidime Cefpirome
1->64 0.5-32 2->64 4->64 2-64 16-> 128 0.25->32
32 8 16 >64 32 128 >32
>64 32 >64 >64 64 >128 >32
1->64 1->64 4->64 2->64 4->64 0.5-> 128 0.5->32
16 8 16 16 16 8 8
>64 >64 >64 >64 >64 >128 >32
Cefepime Piperacillin Gentamicin Acinetobacter calcoaceticus (30)
GR69153 E-1040 Ceftazidime
Cefpirome Cefepime Piperacillin Gentamicin
2 0.5 1 0.06
1 >128 >16
Continued on following page
262
ANTIMICROB. AGENTS CHEMOTHER.
CHIN ET AL. TABLE 1-Continued
(p.g/ml)a 50%
90%
0.015-0.12 0.12-0.25 0.25-1 0.015-0.25 0.015-0.25 0.25-> 128 0.5-4
0.03 0.12 0.5 0.06 0.06 2 1
0.06 0.25 0.5 0.12 0.12 >128 4
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-0.12 0.06-0.5 0.06-1 0.015-0.5 0.015-1 0.25->128 0.25-8
0.06 0.12 0.12 0.06 0.06 2 2
0.12 0.12 0.5 0.12 0.12 >128 4
Aeromonas spp. (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-8 0.015-4 0.12-4 0.015-2 0.015-8 1->128 0.5-8
0.25 0.06 0.25 0.06 0.06 8 1
2 0.25 2 0.25 0.5 64 2
Yersinia enterocolitica (10)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.03-0.25 0.03-2 0.12-8 0.03-0.12 0.03-0.25 0.5-16 0.12-2
0.03 0.06 0.12 0.03 0.03 2 0.5
0.25 0.25
GR69153 E-1040 Ceftazidime Cefpirome
Cefepime Piperacillin Gentamicin
0.06-2 0.12-2 0.06-2 0.06-0.12 0.06-1 0.25->128 2-8
0.12 0.25 0.12 0.06 0.12 2 4
0.25 1 0.25 0.12 0.25 >128
Moraxella catarrhalis (22)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-0.5 0.06-0.5 0.06-0.5 0.06-0.5 0.12-0.5 0.12-2 0.5-8
0.12 0.12 0.12 0.12 0.2 0.5 4
0.5 0.5 0.25 0.5 0.5 2 8
Neisseria gonorrhoeae (10)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-0.5 0.015-0.5 0.015-0.5 0.015-0.5 0.015-0.5 0.06->32 0.25-2
0.06 0.06 0.06 0.06 0.06 >32 0.5
0.12 0.12 0.12 0.12 0.12 >32 2
Staphylococcus aureus, methicillin susceptible (20)
GR69153 E-1040 Ceftazidime Cefpirome
4->32
8 16 8 1 4 8 4
16 16 16 2 8 64 16
8->64
16
32
Antimicrobial Ant agent
~~Range
Salmonella spp. (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
Shigella spp. (15)
Organism (no. of isolates tested)
Haemophilus influenzae (15)
Cefepime Piperacillin Gentamicin
Staphylococcus aureus, methicillin resistant (20)
GR69153
Rag
4-32 8-32 8-32 1-4 2-16 0.5-128
MIC
50%
90%
1
0.06 0.12 4 1
4
Continued on following page
ACTIVITY AND STABILITY OF GR69153
VOL. 35, 1991
263
TABLE 1-Continued MIC (p.g/ml)a
Organism (no. of isolates tested)
Antimicrobial agent
Range
Coagulase-negative staphylococci, methicillin susceptible (10)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.5->64 4-16 4-32 0.25-16 0.5-64 1-64 0.5->32
0.5 4 8 0.5 1 16 4
8 16 32 2 8 64 >32
Coagulase-negative staphylococci, methicillin resistant (10)
GR69153
0.5->64
0.5
8
Streptococcus pyogenes (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-2 0.015-1 0.015-0.5 0.015-0.03 0.015-0.12 0.015-0.12 0.25-8
0.12 0.015 0.015 0.015 0.015 0.06 1
5 0.25 0.25 0.015 0.03 0.12 2
Streptococcus agalactiae (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-0.5 0.015-0.5 0.12-0.5 0.015-0.06 0.015-0.12 0.015-0.25 0.5-4
0.5 0.12 0.25 0.015 0.03 0.06 0.5
0.5 0.5 0.5 0.03 0.06 0.12 4
Streptococcus groups C, F, and G (22)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-1 0.015-2 0.12-4 0.015-0.25 0.015-0.5 0.12 1-4
0.25 0.03 0.25 0.015 0.03 0.12 2
0.5 0.12 0.25 0.12 0.25 0.12 4
Streptococcus pneumoniae (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-1 0.12-2 0.12-4 0.015-0.5 0.015-0.5 0.06-0.12 2-16
0.25 0.25 0.25 0.015 0.03 0.06 8
0.5 1 2 0.12 0.25 0.06 16
Enterococcus faecalis (22)
GR69153 E-1040
Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin Listeria monocytogenes (15)
GR69153 E-1040 Ceftazidime Cefpirome
Cefepime Piperacillin Gentamicin
Bacteroides fragilis (15)
GR69153 E-1040 Ceftazidime
Cefpirome Cefepime Piperacillin Gentamicin
50%
90%
16->64 32->64 64->64 2->64 16->64 2-8 0.5->32
>64 >64 >64 4 32 2 >32
>64 >64 >64 >64 >64 4 >32
2->64 32->64 32->64 1-32 4->64 0.12-4 0.12-4
>64 >64 >64 8 32 2 1
>64 >64 >64 32 64 4 4
0.25->32 32->64 32->64 16-32 8->64 2->128 >32
0.25 >64 >64 32 32 16 >32
8 >64 >64 32 >64 >128 >32
Continued on following page
264
CHIN ET AL.
ANTIMICROB. AGENTS CHEMOTHER. TABLE 1-Continued
Organism (no. of isolates tested)
Antimicrobial agent agent
(15)'
GR69153 E-1040 Ceftazidime Cefpirome Cefepime
Bacteroides spp.
Rag
Piperacillin Gentamicin Clostridium spp. (19)c
MIC
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
(,jLg/ml)a 50%
50%°
90%
0.25->128 32->64 32->64 16-32 8->32 1->128 >32
0.25 >64 >64 32 32 32 >32
32 >64 >64 32 >32 >128 >32
0.5->16 0.5-8 0.5->16
1 2 4 1 1 0.12 >32
8 8 >16 >16 8 1 >32
~~~Range
0.25->16
0.5-> 16 0.06-1 >32
50% and 90%, MIC for 50 and 90o of isolates, respectively. Includes B. melaninogenicus (n = 5), B. bivius (n = 3), B. ovatus (n = 3), and B. thetaiotaomicron (n c Includes C. perfringens (n = 10), C. septicum (n = 3), C. ramsum (n = 3), and C. difficile (n = 3).
90S
a
b
determined by plating 0.01-ml samples from clear 1-ml tubes onto antibiotic-free agar plates. The MBC was the concentration at which there was a 99.9% reduction in CFU based on the CFU in original samples, as determined by the method of Pearson et al. (5). For determination of pH effects, the pH of Mueller-Hinton agar was adjusted with hydrochloric acid or sodium hydroxide to the desired levels. The presence of 1-lactamases in isolates was determined by the nitrocefin assay. The P-lactamases used to determine purified compounds, stability were either purified or partially . . n as described previously (4). Stability to ,-lactamases was' determined by a previously described spectrophotometric assay (4).
RESULTS The activity of GR69153 compared with those of the other agents is shown in Table 1. GR69153 inhibited 50% of the members of the family Enterobacteriaceae at .1 ,ug/ml. It inhibited 90% of Escherichia coli, Klebsiella oxytoca, Citrobacter diversus, Proteus mirabilis, Proteus vulgaris, and Salmonella and Shigella species at .0.25 p.g/ml. Against these species, GR69153 had activity comparable to those of ceftazidime, cefpirome, cefepime, and E-1040; and it inhibited organisms resistant to piperacillin and gentamicin. For some isolates of Klebsiella pneumoniae, Enterobacter species, and Citrobacter freundii, as well as Morganella morganii, MICs were >8 ,ug/ml. GR69153 had activity similar to
=
4).
that of ceftazidime but was less active than E-1040, cefpirome, and cefepime against Enterobacter species and Citrobacterfreundii. GR69153 was less active than the other cephalosporins against Morganella morganii. For approximately 10% of the isolates of Proteus rettgeri, Providencia stuartii, and Serratia marcescens, GR69153 MICs were 2
jig/ml. GR69153 inhibited 90% of Pseudomonas aeruginosa at .2
. lates
i
g
and p
om the cefazidindrefis and isolates for which the ceftazidime and cefpilrome ~~~~lates andisolaesfrwich
MICs
were >16 jig/ml, and had activity similar to that of E-1040. For most Pseudomonas cepacia, Xanthomonas maltophilia, and Acinetobacter calcoaceticus isolates, MICs were >16 jigIml. GR69153 inhibited Haemophilus influenzae and Moraxella catarrhalis at concentrations similar to those of the other agents. Penicillinase-producing Neisseria gonorrhoeae was inhibited by .0.12 p.g/ml. Against Staphylococcus aureus GR69153 had activity similar to those of ceftazidime and E-1040 (MIC for 90% of strains tested, 16 ,ug/ml). It was less active than cefpirome and did not inhibit methicillin-resistant staphylococci. GR69153 was less active than the other agents against group A streptococci, but 90% of the isolates were inhibited by 0.5 ,uglml, as were group B, C, F, and 0 streptococci. GR69153 had activity similar to that of ceftazidime against Streptococcus pneumoniae but was less active than cefpirome, cefepime, and piperacillin. Similar to the other cephalospo-
TABLE 2. Effect of inoculum size on MICs of GR69153'
Organism' Escherichia coli Klebsiella pneumoniae Citrobacter diversus Serratia marcescens Pseudomonas aeruginosa Staphylococcus aureus a
b
Determined on Mueller-Hinton agar. Five isolates of each species were tested.
Geometric
mean
MIC
(p.g/ml [range]) at CFU of:
103
105
107
0.04 (0.03-0.06) 0.05 (0.03-0.06) 0.03 (0.03-0.03) 0.31 (0.12-0.5) 0.22 (0.12-0.5) 4 (4-4)
0.04 (0.03-0.06) 0.05 (0.03-0.12) 0.05 (0.03-0.12) 0.42 (0.25-1) 0.33 (0.25-0.5) 8 (8-8)
1 (1-1) 0.5 (0.5-0.5) 0.25 (0.25-0.25) 4 (4-4) 2.3 (1-4) 16 (16-16)
265
ACTIVITY AND STABILITY OF GR69153
VOL. 35, 1991
TABLE 3. Effect of pH on MICs of GR69153 7.5
Geometric mean MIC (jig/ml [range]) at pH of: 6.5
5.5
0.03 (0.03-0.03) 0.09 (0.06-0.12) 0.03 (0.03-0.03) 0.25 (0.12-0.5) 0.5 (0.25-1) 6.96 (4-8)
0.05 (0.03-0.12) 0.09 (0.06-0.12) 0.06 (0.03-0.25) 0.41 (0.25-0.5) 0.66 (0.5-1) 6.06 (4-8)
0.16 (0.12-0.25) 0.5 (0.5-0.5) 0.25 (0.25-0.25) 1 (1-1) 6.96 (4-64) 5.3 (4-8)
Organisma
Escherichia coli Klebsiella pneumoniae Citrobacter diversus Serratia marcescens Pseudomonas aeruginosa Staphylococcus aureus a Five isolates of each species were tested.
rins, GR69153 failed to inhibit Enterococcus faecalis and Listeria monocytogenes. GR69153 was more active than the other cephalosporins against Bacteroides fragilis and other Bacteroides species, inhibiting most Bacteroides at .8 ug/Iml, with the exception of Bacteroides thetaiotaomicron, for which the MIC was >32 ,ug/ml. Peptostreptococci and peptococci (five isolates each; data not shown) were inhibited by .0.25 pg/ml. Clostridium perfringens was inhibited by 0.5 .g/nml, but for Clostridium difficile the GR69153 MIC was 8 ,ug/ml. Propionibacterium species (four isolates; data not shown) were inhibited by .0.25 ,ug of GR69153 per ml. Effect of growth conditions. The effect of inoculum size is shown in Table 2. At 107 CFU there was an appreciable increase in the MIC compared with the MIC at 105 CFU for members of the family Enterobacteriaceae and Pseudomonas aeruginosa. GR69153 was more active at pH 7.5 than it was at pH 5.5 (Table 3), with, for example, the mean MIC for Pseudomonas aeruginosa increasing from 0.5 ,ug/ml at pH 7.5 to 7 pLg/ml at pH 5.5 and increasing for one isolate from 1 to 64 ,ug/ml. There was a similar effect of pH on Serratia marcescens. There was no effect on the MIC for Staphylococcus aureus. The GR69153 MBC for the members of the family Enterobacteriaceae tested increased minimally compared with the MIC, but the GR69153 MBC for Pseudomo-
nas aeruginosa was 8- to 16-fold greater than the MIC. The GR69153 MBC determined in 50% human serum for Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae (one isolate each) was within a dilution of the MBC determined in Mueller-Hinton broth. The activity of GR69153 against Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus (five isolates of each species tested) was similar in room air and in the anaerobic environment of the GasPak jar system. ,I-Lactamase stability. The activity of GR69153 compared with those of the other agents against organisms with charthe excepacterized of-lactamases is shown in Table 4. With ins the ceperobacter loaceowhich c tio tion of Enterobacter cloacae, which contains the cephalosporinase P99, Escherichia coli, which contains TEM-9, and a Citrobacter freundii isolate which possessed TEM-7 as well as its own chromosomal 1-lactamase, all organisms were inhibited by 64 0.12 1 1 0.5 0.5 0.5 1 0.12 0.12 0.12
0.25 0.25 8 8 32 >64 0.25 0.25 8 8 8 >64 0.25 4 4 4 8 4 2 8 4 0.12
0.03 0.03 >64 0.06 0.25 8 0.25 0.12 >64 16 1 >64 0.12 16 32 >64 >64 >64 >64 >64 >64 0.03
CHIN ET AL.
266
ANTIMICROB. AGENTS CHEMOTHER. TABLE 5. Stability of GR69153 to P-lactamases
TEM-1 TEM-2 TEM-3 TEM-9 SHV-1 Kl P99
PSE-1 PSE-4 Sabath-Abraham BRO-1 PC
Relative hydrolysis rate
Source
Enzyme
Escherichia coli Escherichia coli Klebsiella pneumoniae Escherichia coli Klebsiella pneumoniae Klebsiella oxytoca Enterobacter cloacae Pseudomonas aeruginosa Pseudomonas aeruginosa Pseudomonas aeruginosa Moraxella catarrhalis Staphylococcus aureus Morganella morganii
of':
GR69153
Cefotaxime
Ceftazidime
p.
Vol. 35, No. 2
259-266
In Vitro Activity and 3-Lactamase Stability of GR69153, a New Long-Acting Cephalosporin NAI-XUN CHIN,' JIAN-WEI GU,' WEI FANG,' AND HAROLD C. NEU' 2* Departments of Medicine' and Pharmacology,2 College of Physicians and Surgeons, Columbia University, New York, New York 10032 Received 11 September 1990/Accepted 20 November 1990
GR69153, a new parenteral cephalosporin, inhibited 90% of Escherichia coli, Klebsiella oxytoca, Proteus mirabilis, Citrobacter diversus, shigellae, and salmonellae at 32 ,ug/ml, and cefpirome and cefepime were the most active agents against these species. Pseudomonas aeruginosa was highly susceptible to GR69153, and for this organism the MIC90 was 128 0.5-4
0.06 0.12 0.25 0.03 0.03 2 1
0.25 0.25 0.5 0.12 0.12 >128 2
Klebsiella pneumoniae (30)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015->16 0.06->16 0.12->16 0.015->32 0.015->32 1->128 0.5-8
0.06 0.12 0.25 0.06 0.06 8 1
1 0.5 1 1 1 >128 4
Klebsiella oxytoca (20)
GR69153 E-1040 Ceftazidime
Cefpirome Cefepime Piperacillin Gentamicin
0.015-4 0.06-2 0.12-2 0.015-1 0.015-1 2->128 0.5-1
0.06 0.12 0.25 0.06 0.06 8 0.5
0.12 0.5 0.5 0.25 0.25 >128 1
Enterobacter aerogenes (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-32 0.06-0.5 0.12->32 0.015-1 0.015-0.5 2-128 0.25-1
0.12 0.12 0.5 0.12 0.25 4 0.5
8 0.25 32 0.5 0.5 64 1
Enterobacter cloacae (30)
GR69153 E-1040 Ceftazidime Cefpirome
Cefepime Piperacillin Gentamicin
0.015->32 0.015-4 0.015->32 0.015-16 0.015-4 32->256 0.5->16
0.25 0.12 4 0.25 0.25 128 1
>32 2 >32 16 2 >128 >16
Citrobacterfreundii (30)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015->32 0.015->32 0.015->32 0.015->32 0.015-16 1->128 0.25->16
1 0.12 1 0.12 0.06 4 1
>32 4 >32 8 8 >128 16
Citrobacter diversus (15)
GR69153 E-1040 Ceftazidime
0.015-0.06 0.015-0.12 0.015-0.25 0.015-0.06 0.015-0.06
0.06 0.12 0.12 0.06 0.06 8 0.5
0.06 0.12 0.25 0.06 0.06 32 2
0.06 0.25 0.06 0.015 0.03 0.5 1
0.06 0.5 0.12 0.03 0.06 1 16
0.5 0.06 0.06
8 0.06 0.5
0.015
0.015
Cefpirome Cefepime Piperacillin Gentamicin Proteus mirabilis (20)
GR69153 E-1040
Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
Morganella morganii (10)
GR69153 E-1040 Ceftazidime
Cefpirome
4->128 0.25-4 0.06-0.25 0.12-1 0.06-0.25 0.015-0.5 0.03-0.5 0.25-1 0.5->16
0.06-8 0.06-0.25 0.015-1 0.015-0.03
Continued on following page
ACTIVITY AND STABILITY OF GR69153
VOL. 35, 1991
261
TABLE 1-Continued Organism (no. of isolates tested)
MIC Antimicrobial (gng/ml)a agent Range 50%
90%
0.015 8 1
Cefepime Piperacillin Gentamicin
0.015-0.03 0.12-32 0.5-1
0.015 2 0.5
Proteus vulgaris (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-0.12 0.12-1 0.06-0.25 0.03-1 0.015-0.12 0.25->128 0.12->16
0.06 0.12 0.06 0.12 0.03 8 0.5
0.12 0.5 0.12 0.5 0.06 >128 2
Proteus rettgeri (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-2 0.06-0.5 0.06-1 0.06-0.5 0.06-0.5 0.25->128 1->16
2 0.25 0.25 0.25 0.25 0.5 2
2 0.5 0.5 0.5 0.5 >128 16
Providencia stuartii (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-4 0.06-0.5 0.06-1 0.06-0.25 0.06-4 0.25->128 2->16
0.06 0.06 0.12 0.06 0.06 8 8
Serratia marcescens (30)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-4 0.12-8 0.12-16 0.06-4 0.06-4 1->128 1->16
0.25 0.25 0.25 0.12 0.12 16 2
1 4 1 0.5 1 >128 8
Pseudomonas aeruginosa (45)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-8 0.25-16 1->32 2->32 1-16 2->128 2->16
0.5 0.5 2 4 2 32 4
2 1 32 32 8 >128 >16
Pseudomonas cepacia (17)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.25->64 0.12-64 2->64 0.12->64 0.12-64 2->128 0.25->16
16 32 16 8 8 8 2
>64 64 >64 >64 64 >128 >32
Xanthomonas maltophilia (12)
GR69153 E-1040 Ceftazidime Cefpirome
1->64 0.5-32 2->64 4->64 2-64 16-> 128 0.25->32
32 8 16 >64 32 128 >32
>64 32 >64 >64 64 >128 >32
1->64 1->64 4->64 2->64 4->64 0.5-> 128 0.5->32
16 8 16 16 16 8 8
>64 >64 >64 >64 >64 >128 >32
Cefepime Piperacillin Gentamicin Acinetobacter calcoaceticus (30)
GR69153 E-1040 Ceftazidime
Cefpirome Cefepime Piperacillin Gentamicin
2 0.5 1 0.06
1 >128 >16
Continued on following page
262
ANTIMICROB. AGENTS CHEMOTHER.
CHIN ET AL. TABLE 1-Continued
(p.g/ml)a 50%
90%
0.015-0.12 0.12-0.25 0.25-1 0.015-0.25 0.015-0.25 0.25-> 128 0.5-4
0.03 0.12 0.5 0.06 0.06 2 1
0.06 0.25 0.5 0.12 0.12 >128 4
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-0.12 0.06-0.5 0.06-1 0.015-0.5 0.015-1 0.25->128 0.25-8
0.06 0.12 0.12 0.06 0.06 2 2
0.12 0.12 0.5 0.12 0.12 >128 4
Aeromonas spp. (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-8 0.015-4 0.12-4 0.015-2 0.015-8 1->128 0.5-8
0.25 0.06 0.25 0.06 0.06 8 1
2 0.25 2 0.25 0.5 64 2
Yersinia enterocolitica (10)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.03-0.25 0.03-2 0.12-8 0.03-0.12 0.03-0.25 0.5-16 0.12-2
0.03 0.06 0.12 0.03 0.03 2 0.5
0.25 0.25
GR69153 E-1040 Ceftazidime Cefpirome
Cefepime Piperacillin Gentamicin
0.06-2 0.12-2 0.06-2 0.06-0.12 0.06-1 0.25->128 2-8
0.12 0.25 0.12 0.06 0.12 2 4
0.25 1 0.25 0.12 0.25 >128
Moraxella catarrhalis (22)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-0.5 0.06-0.5 0.06-0.5 0.06-0.5 0.12-0.5 0.12-2 0.5-8
0.12 0.12 0.12 0.12 0.2 0.5 4
0.5 0.5 0.25 0.5 0.5 2 8
Neisseria gonorrhoeae (10)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.015-0.5 0.015-0.5 0.015-0.5 0.015-0.5 0.015-0.5 0.06->32 0.25-2
0.06 0.06 0.06 0.06 0.06 >32 0.5
0.12 0.12 0.12 0.12 0.12 >32 2
Staphylococcus aureus, methicillin susceptible (20)
GR69153 E-1040 Ceftazidime Cefpirome
4->32
8 16 8 1 4 8 4
16 16 16 2 8 64 16
8->64
16
32
Antimicrobial Ant agent
~~Range
Salmonella spp. (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
Shigella spp. (15)
Organism (no. of isolates tested)
Haemophilus influenzae (15)
Cefepime Piperacillin Gentamicin
Staphylococcus aureus, methicillin resistant (20)
GR69153
Rag
4-32 8-32 8-32 1-4 2-16 0.5-128
MIC
50%
90%
1
0.06 0.12 4 1
4
Continued on following page
ACTIVITY AND STABILITY OF GR69153
VOL. 35, 1991
263
TABLE 1-Continued MIC (p.g/ml)a
Organism (no. of isolates tested)
Antimicrobial agent
Range
Coagulase-negative staphylococci, methicillin susceptible (10)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.5->64 4-16 4-32 0.25-16 0.5-64 1-64 0.5->32
0.5 4 8 0.5 1 16 4
8 16 32 2 8 64 >32
Coagulase-negative staphylococci, methicillin resistant (10)
GR69153
0.5->64
0.5
8
Streptococcus pyogenes (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.06-2 0.015-1 0.015-0.5 0.015-0.03 0.015-0.12 0.015-0.12 0.25-8
0.12 0.015 0.015 0.015 0.015 0.06 1
5 0.25 0.25 0.015 0.03 0.12 2
Streptococcus agalactiae (15)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-0.5 0.015-0.5 0.12-0.5 0.015-0.06 0.015-0.12 0.015-0.25 0.5-4
0.5 0.12 0.25 0.015 0.03 0.06 0.5
0.5 0.5 0.5 0.03 0.06 0.12 4
Streptococcus groups C, F, and G (22)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-1 0.015-2 0.12-4 0.015-0.25 0.015-0.5 0.12 1-4
0.25 0.03 0.25 0.015 0.03 0.12 2
0.5 0.12 0.25 0.12 0.25 0.12 4
Streptococcus pneumoniae (20)
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
0.12-1 0.12-2 0.12-4 0.015-0.5 0.015-0.5 0.06-0.12 2-16
0.25 0.25 0.25 0.015 0.03 0.06 8
0.5 1 2 0.12 0.25 0.06 16
Enterococcus faecalis (22)
GR69153 E-1040
Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin Listeria monocytogenes (15)
GR69153 E-1040 Ceftazidime Cefpirome
Cefepime Piperacillin Gentamicin
Bacteroides fragilis (15)
GR69153 E-1040 Ceftazidime
Cefpirome Cefepime Piperacillin Gentamicin
50%
90%
16->64 32->64 64->64 2->64 16->64 2-8 0.5->32
>64 >64 >64 4 32 2 >32
>64 >64 >64 >64 >64 4 >32
2->64 32->64 32->64 1-32 4->64 0.12-4 0.12-4
>64 >64 >64 8 32 2 1
>64 >64 >64 32 64 4 4
0.25->32 32->64 32->64 16-32 8->64 2->128 >32
0.25 >64 >64 32 32 16 >32
8 >64 >64 32 >64 >128 >32
Continued on following page
264
CHIN ET AL.
ANTIMICROB. AGENTS CHEMOTHER. TABLE 1-Continued
Organism (no. of isolates tested)
Antimicrobial agent agent
(15)'
GR69153 E-1040 Ceftazidime Cefpirome Cefepime
Bacteroides spp.
Rag
Piperacillin Gentamicin Clostridium spp. (19)c
MIC
GR69153 E-1040 Ceftazidime Cefpirome Cefepime Piperacillin Gentamicin
(,jLg/ml)a 50%
50%°
90%
0.25->128 32->64 32->64 16-32 8->32 1->128 >32
0.25 >64 >64 32 32 32 >32
32 >64 >64 32 >32 >128 >32
0.5->16 0.5-8 0.5->16
1 2 4 1 1 0.12 >32
8 8 >16 >16 8 1 >32
~~~Range
0.25->16
0.5-> 16 0.06-1 >32
50% and 90%, MIC for 50 and 90o of isolates, respectively. Includes B. melaninogenicus (n = 5), B. bivius (n = 3), B. ovatus (n = 3), and B. thetaiotaomicron (n c Includes C. perfringens (n = 10), C. septicum (n = 3), C. ramsum (n = 3), and C. difficile (n = 3).
90S
a
b
determined by plating 0.01-ml samples from clear 1-ml tubes onto antibiotic-free agar plates. The MBC was the concentration at which there was a 99.9% reduction in CFU based on the CFU in original samples, as determined by the method of Pearson et al. (5). For determination of pH effects, the pH of Mueller-Hinton agar was adjusted with hydrochloric acid or sodium hydroxide to the desired levels. The presence of 1-lactamases in isolates was determined by the nitrocefin assay. The P-lactamases used to determine purified compounds, stability were either purified or partially . . n as described previously (4). Stability to ,-lactamases was' determined by a previously described spectrophotometric assay (4).
RESULTS The activity of GR69153 compared with those of the other agents is shown in Table 1. GR69153 inhibited 50% of the members of the family Enterobacteriaceae at .1 ,ug/ml. It inhibited 90% of Escherichia coli, Klebsiella oxytoca, Citrobacter diversus, Proteus mirabilis, Proteus vulgaris, and Salmonella and Shigella species at .0.25 p.g/ml. Against these species, GR69153 had activity comparable to those of ceftazidime, cefpirome, cefepime, and E-1040; and it inhibited organisms resistant to piperacillin and gentamicin. For some isolates of Klebsiella pneumoniae, Enterobacter species, and Citrobacter freundii, as well as Morganella morganii, MICs were >8 ,ug/ml. GR69153 had activity similar to
=
4).
that of ceftazidime but was less active than E-1040, cefpirome, and cefepime against Enterobacter species and Citrobacterfreundii. GR69153 was less active than the other cephalosporins against Morganella morganii. For approximately 10% of the isolates of Proteus rettgeri, Providencia stuartii, and Serratia marcescens, GR69153 MICs were 2
jig/ml. GR69153 inhibited 90% of Pseudomonas aeruginosa at .2
. lates
i
g
and p
om the cefazidindrefis and isolates for which the ceftazidime and cefpilrome ~~~~lates andisolaesfrwich
MICs
were >16 jig/ml, and had activity similar to that of E-1040. For most Pseudomonas cepacia, Xanthomonas maltophilia, and Acinetobacter calcoaceticus isolates, MICs were >16 jigIml. GR69153 inhibited Haemophilus influenzae and Moraxella catarrhalis at concentrations similar to those of the other agents. Penicillinase-producing Neisseria gonorrhoeae was inhibited by .0.12 p.g/ml. Against Staphylococcus aureus GR69153 had activity similar to those of ceftazidime and E-1040 (MIC for 90% of strains tested, 16 ,ug/ml). It was less active than cefpirome and did not inhibit methicillin-resistant staphylococci. GR69153 was less active than the other agents against group A streptococci, but 90% of the isolates were inhibited by 0.5 ,uglml, as were group B, C, F, and 0 streptococci. GR69153 had activity similar to that of ceftazidime against Streptococcus pneumoniae but was less active than cefpirome, cefepime, and piperacillin. Similar to the other cephalospo-
TABLE 2. Effect of inoculum size on MICs of GR69153'
Organism' Escherichia coli Klebsiella pneumoniae Citrobacter diversus Serratia marcescens Pseudomonas aeruginosa Staphylococcus aureus a
b
Determined on Mueller-Hinton agar. Five isolates of each species were tested.
Geometric
mean
MIC
(p.g/ml [range]) at CFU of:
103
105
107
0.04 (0.03-0.06) 0.05 (0.03-0.06) 0.03 (0.03-0.03) 0.31 (0.12-0.5) 0.22 (0.12-0.5) 4 (4-4)
0.04 (0.03-0.06) 0.05 (0.03-0.12) 0.05 (0.03-0.12) 0.42 (0.25-1) 0.33 (0.25-0.5) 8 (8-8)
1 (1-1) 0.5 (0.5-0.5) 0.25 (0.25-0.25) 4 (4-4) 2.3 (1-4) 16 (16-16)
265
ACTIVITY AND STABILITY OF GR69153
VOL. 35, 1991
TABLE 3. Effect of pH on MICs of GR69153 7.5
Geometric mean MIC (jig/ml [range]) at pH of: 6.5
5.5
0.03 (0.03-0.03) 0.09 (0.06-0.12) 0.03 (0.03-0.03) 0.25 (0.12-0.5) 0.5 (0.25-1) 6.96 (4-8)
0.05 (0.03-0.12) 0.09 (0.06-0.12) 0.06 (0.03-0.25) 0.41 (0.25-0.5) 0.66 (0.5-1) 6.06 (4-8)
0.16 (0.12-0.25) 0.5 (0.5-0.5) 0.25 (0.25-0.25) 1 (1-1) 6.96 (4-64) 5.3 (4-8)
Organisma
Escherichia coli Klebsiella pneumoniae Citrobacter diversus Serratia marcescens Pseudomonas aeruginosa Staphylococcus aureus a Five isolates of each species were tested.
rins, GR69153 failed to inhibit Enterococcus faecalis and Listeria monocytogenes. GR69153 was more active than the other cephalosporins against Bacteroides fragilis and other Bacteroides species, inhibiting most Bacteroides at .8 ug/Iml, with the exception of Bacteroides thetaiotaomicron, for which the MIC was >32 ,ug/ml. Peptostreptococci and peptococci (five isolates each; data not shown) were inhibited by .0.25 pg/ml. Clostridium perfringens was inhibited by 0.5 .g/nml, but for Clostridium difficile the GR69153 MIC was 8 ,ug/ml. Propionibacterium species (four isolates; data not shown) were inhibited by .0.25 ,ug of GR69153 per ml. Effect of growth conditions. The effect of inoculum size is shown in Table 2. At 107 CFU there was an appreciable increase in the MIC compared with the MIC at 105 CFU for members of the family Enterobacteriaceae and Pseudomonas aeruginosa. GR69153 was more active at pH 7.5 than it was at pH 5.5 (Table 3), with, for example, the mean MIC for Pseudomonas aeruginosa increasing from 0.5 ,ug/ml at pH 7.5 to 7 pLg/ml at pH 5.5 and increasing for one isolate from 1 to 64 ,ug/ml. There was a similar effect of pH on Serratia marcescens. There was no effect on the MIC for Staphylococcus aureus. The GR69153 MBC for the members of the family Enterobacteriaceae tested increased minimally compared with the MIC, but the GR69153 MBC for Pseudomo-
nas aeruginosa was 8- to 16-fold greater than the MIC. The GR69153 MBC determined in 50% human serum for Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae (one isolate each) was within a dilution of the MBC determined in Mueller-Hinton broth. The activity of GR69153 against Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus (five isolates of each species tested) was similar in room air and in the anaerobic environment of the GasPak jar system. ,I-Lactamase stability. The activity of GR69153 compared with those of the other agents against organisms with charthe excepacterized of-lactamases is shown in Table 4. With ins the ceperobacter loaceowhich c tio tion of Enterobacter cloacae, which contains the cephalosporinase P99, Escherichia coli, which contains TEM-9, and a Citrobacter freundii isolate which possessed TEM-7 as well as its own chromosomal 1-lactamase, all organisms were inhibited by 64 0.12 1 1 0.5 0.5 0.5 1 0.12 0.12 0.12
0.25 0.25 8 8 32 >64 0.25 0.25 8 8 8 >64 0.25 4 4 4 8 4 2 8 4 0.12
0.03 0.03 >64 0.06 0.25 8 0.25 0.12 >64 16 1 >64 0.12 16 32 >64 >64 >64 >64 >64 >64 0.03
CHIN ET AL.
266
ANTIMICROB. AGENTS CHEMOTHER. TABLE 5. Stability of GR69153 to P-lactamases
TEM-1 TEM-2 TEM-3 TEM-9 SHV-1 Kl P99
PSE-1 PSE-4 Sabath-Abraham BRO-1 PC
Relative hydrolysis rate
Source
Enzyme
Escherichia coli Escherichia coli Klebsiella pneumoniae Escherichia coli Klebsiella pneumoniae Klebsiella oxytoca Enterobacter cloacae Pseudomonas aeruginosa Pseudomonas aeruginosa Pseudomonas aeruginosa Moraxella catarrhalis Staphylococcus aureus Morganella morganii
of':
GR69153
Cefotaxime
Ceftazidime
