In vitro susceptibilities of common pediatric pathogens to LY163892.

S Shelton and J D Nelson Antimicrob. Agents Chemother. 1988, 32(2):268. DOI: 10.1128/AAC.32.2.268.

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ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Feb. 1988, p. 268-270 0066-4804/88/020268-03$02.00/0 Copyright C) 1988, American Society for Microbiology

Vol. 32, No. 2

In Vitro Susceptibilities of Common Pediatric Pathogens to LY163892

SHARON SHELTON AND JOHN D. NELSON* Department of Pediatrics, The University of Texas Health Science Center at Dallas, 5323 Harry Hines Boulevard, Dallas, Texas 75235
Received 31 August 1987/Accepted 30 November 1987

LY163892 is a carbacephem antibiotic for oral administration with an antibacterial spectrum similar to that of cefaclor and amoxicillin-clavulanic acid. It has greater stability than cefaclor and greater activity against ,3-lactamase-producing Haemophilus influenzae and Escherichia coli. LY163892 is less active than amoxicillin against streptococci and less active than amoxicillin-clavulanic acid against Branhamella catarrhalis but comparable against other pathogens.

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LY163892 is a carbacephem synthetic antibiotic for oral administration. Its chemical name is 7-[D-(aminophenylacetyl) amino]-3-chloro-8-oxo-1-azabicyclo[4.2.0]oct-2-ene2-carboxylic acid. Structurally it is similar to cefaclor, with the major difference being the substitution of a sulfur molecule for carbon at position 1 in the dihydrothiazine ring. In preliminary tests by the manufacturer, it had a spectrum of in vitro activity comparable to that of cefaclor. The purpose of this study was to examine the in vitro activity of LY163892 against common bacterial pathogens that cause upper and lower respiratory tract infections, urinary tract infections, and infections of the skin and skin structures in comparison with cefaclor, amoxicillin, and amoxicillin-clavulanic acid. Laboratory-standard powders were obtained from the manufacturers. The Streptococcus pyogenes isolates were from throat (n = 52) and middle ear fluid (n = 11) cultures. The Escherichia coli strains were from urine cultures. Branhamella catarrhalis, Haemophilus influenzae, Streptococcus pneumoniae, and Staphylococcus aureus strains (all methicillin susceptible) were mainly from middle ear fluid cultures. The bacterial isolates had been collected from clinical specimens between 1980 and 1986. Mueller-Hinton agar (B. catarrhalis, S. aureius, E. coli), 5% sheep blood agar (S. pyogenes, S. pneumoniae), and chocolate agar (H. influenzae) were used for agar dilution studies. Twofold dilutions of antibiotic from 8 to 0.03 ,ug/ml were added to the liquefied agar. Amoxicillin and clavulanic acid were in a 4:1 ratio, with the concentration expressed as the amoxicillin component. Seed cultures of the bacterial strains were grown overnight at 37C in appropriate broth media and diluted in Mueller-Hinton broth so that the prongs of a Steers-Foltz replicator (which deliver 0.007 ml) would deliver an inoculum of 104 to 105 CFU. Actual colony counts were done on each strain tested and ranged from 9 x 103 to 9 x 105, with most being 105. The agar plates were incubated at 37C for 18 h. The MIC was the smallest amount of drug resulting in no growth or fewer than 10 colonies. Disk testing was done with a 30-p.g LY163892 disk by using the National Committee for Clinical Laboratory Standards modification of the Kirby-Bauer method (2). According to the manufacturer, a zone size of <14 mm indicates
*

TABLE 1. Effect of bacterial inoculum size on susceptibility

Concn (,ug/ml) at inoculum size (CFU):

Organism
MIC

102

104

106

MBC MIC MBC MIC MBC

B. catarrhalis, ,B-lactamase positive (N86-0121E) B. catarrhalis, P-lactamase

0.5

1.0

0.5 1.0 8.0 >8.0 0.25 0.5 0.5

0.12

0.12 0.12 0.12 0.25

negative (N86-O111E) H. influenzae, ,B-lactamase positive (N84-047-0) H. influenzae, P-lactamase negative (N84-014-0) S. aureus, 1-lactamase positive (N86-0204-E) S. pyogenes S. pneumoniae, relatively resistant (N86-0188E) S. pneumoniae, penicillin susceptible (N86-0189E)

<0.008 0.5

0.06 0.5 0.5

0.016 0.016 0.12 0.12 0.5

0.5

1.0

1.0 2.0 8.0 >8.0

0.25 2.0

0.25 0.12 0.12 0.25 2.0 2.0 4.0 4.0

0.25 4.0
0.12

0.12

0.12 0.12 0.12 0.12

TABLE 2. Effect of fresh human serum on susceptibility

Concn (pLg/ml) in:

Organism"
MIC

Broth

Broth with 50%

serum

MBC

MIC

MBC

B. catarrhalis, ,3-lactamase

0.5 0.12
0.06

1.0
0.12
0.5

0.25
0.06
0.06

0.25
0.12 0.25 0.25 2.0

positive B. catarrhalis, 3-lactamase negative H. influenzae, 1B-lactamase positive H. influenzae, P-lactamase negative S. aureus, P-lactamase positive S. pyogenes S. pneumoniae, relatively
resistant

0.12 1.0
0.25 2.0

0.12
2.0

0.03
2.0 0.25 4.0 0.25

0.25 4.0
0.12

0.25 4.0
0.25

S. pneumoniae, penicillin

0.12

susceptible
Corresponding author.
268
" Strain identification numbers are the same as in Table 1.

VOL. 32, 1988

NOTES
-

269

8r
E
at
._

~~~vv

TABLE 3. Agar dilution susceptibility test results

V

vW
V 7"l
v

Organism (no. of strains tested)

o

MIC (,g/mi)b Dru rugRange 50

90
1 1

0
0

2_

B. catarrhalis,

c 0

P-lactamase
1
v
4c

positive (40)
0

LY CEF AMOX AMOX-CA

1-

Rage5-29P/
0.5-2

s0.03-2 <0.03->8 s0.03-0.25 s0.03-0.5

.0.03-0.5

0.5 1 1

0.12

4 0.25 0.12 0.25

<0.03 <0.03

.0
C 0

0.5 _
0.2.5

v S. oureus

Str. pneumonioe

cJ

(Relatively Resistant)
a

B. catarrhalis, lactamase negative (8)

0

E
C

Str. pneumoniae

68a
68,

LY CEF AMOX AMOX-CA LY CEF AMOX AMOX-CA

LY CEF AMOX AMOX-CA

0.12 0.12
.0.03 .0.03

s0.03-0.12 s0.03
0.12-0.5 1-8 1->8 <0.03-2
<0.03-1 0.25-8 s0.03-0.5 s0.03-0.5 0.12-1 2->8 2->8 0.05-4 0.25-1 1->8 0.12-0.5 0.25-0.5
0.5-8 1->8 0.12->8 0.12-2

(Penicillin Susceptible)

0.125
0.06

Str. pyogenes

o0

r._

H. influenzae type b, P-lactamase positive (24)

H. influenzae type b, P-lactamase negative (25) H. influenzae NTC,

0.25 0.5 4 4 >8 >8 1 1

0.25 4 0.25 0.25
0.5 8 >8 2

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40lo_ o_ s 0.03 20 lO 30 40 Zone Size (mm) Around 30Etg DISC FIG. 1. LY163892 disk zone diameters and MICs for grampositive bacteria.
L

0.o

0.5 8 0.5 0.5

1

P-lactamase positive (14)

H. influenzae NT,

LY CEF AMOX AMOX-CA

LY CEF AMOX AMOX-CA
LY CEF AMOX AMOX-CA LY CEF AMOX AMOX-CA LY CEF AMOX AMOX-CA
LY CEF AMOX AMOX-CA

>8
>8 2

resistance, 15 to 17 mm indicates intermediate, and -18 mm indicates susceptibility. Initially, a representative strain of each type of organism was tested for inoculum effect and for the effect of human serum on MICs and MBCs determined by the broth microdilution method described by Barry (1). The manufacturer considers strains for which MICs are 8 p,g/ml or less to be susceptible. Haemophilus strains were tested in MuellerHinton broth with supplement C (Difco Laboratories), Branhamella and staphylococcal strains were tested in MuellerHinton broth, and streptococcal strains were tested in Todd-Hewitt broth with 5% sheep erythrocytes. With ino8

P-lactamase negative (31)

S.
aureus

0.5 8 0.5 0.5 2-4 4 4 2

1 >8 0.5 0.5

4

(25)

8 >8 2

S.

pyogenes

(63)

s0.03-0.5 s0.03-0.25
.0.03

s0.03
0.25->8 1->8 0.5->8 0.5->8
81 >82 81,>82 0.51,22 0.51,22

0.06 0.12 0.12 0.12 .0.03 <0.03 s0.03 <0.03 0.5 4 4 4

2 >8 >8 >8

E. coli (51)

E
4
-

S. pneumoniae,

2
C,

penicillin resistant (MIC, >1.0

0
.0
-

p.g/ml) (3)

NA" NA NA NA
0.5 0.5 0.12 0.06

NA NA NA NA 4 1 0.12 0.12

c 0

1I

o0 bgeB400
0.5 _
El

0
._

S. pneumoniae, rel- LY CEF atively resistant AMOX (MIC, 0.1-1.0 AMOX-CA ,ug/ml) (17)

0.12-8 0.25-8 <0.03-0.25 .0.03-0.12

0.25 _ 0.125
< 0.06
10

cotorrhalis v E. co/i
o
8.

18
0

S. pneumoniae,

op8
0

o
._

H. influenzoe

0
1

penicillin susceptible (MIC, <0.1 p.g/ml) (26)

LY CEF AMOX AMOX-CA

<0.03-1 0.12-1 <0.03 <0.03-0.06

1 0.5 0.5 0.5 .0.03 <0.03 <0.03 <0.03

._

40 30 Zone Size (mm) Around 30,4g D)ISC

20

' LY, LY163892; CEF, cefaclor; AMOX, amoxicillin; AMOX-CA, amoxicillin-clavulanic acid. 'b 50% and 90%, MIC for 50 and 90% of strains tested, respectively. c NT, Nontypable. d Subscripts indicate the numbers of strains for which MIC was as indi-

cated.

FIG. 2. LY163892 disk negative bacteria.

zone

diameters and MICs for gram-

e NA,

Not applicable.

270

NOTES

ANTIMICROB. AGENTS CHEMOTHER.

cula of 102, 104, and 106 CFU, results were similar except with P-lactamase-positive B. catarrhalis and S. aureus for which MICs and MBCs increased approximately eightfold with the largest inoculum (Table 1). There was no increase in MIC or MBC in broth containing 50% freshly drawn human serum compared with plain broth, and in the case of ,Blactamase-positive B. catarrhalis the MIC and MBC were two- to fourfold lower in broth with 50% serum (Table 2). Results of agar dilution susceptibility testing results are shown in Table 3. LY163892 exhibited good in vitro activity against all bacterial strains tested except two strains of E. coli that were resistant to 8 ,ug/ml. Those two strains were also resistant to 8 ,ug of the other three drugs tested per ml. In comparison with cefaclor, LY163892 had approximately 16-fold-greater activity against ,-lactamase-positive or -negative H. influenzae, approximately 8-fold-greater activity against E. coli, and comparable activity against the other pathogens. In comparison with amoxicillin and amoxicillin-clavulanic acid, LY163892 was less active against S. pyogenes, S.

pneumoniae, and B. catarrhalis, more active against E. coli, and comparably active against the other pathogens. The 30-,ug LY163892 disk did not discriminate well the various MICs for gram-positive cocci (Fig. 1), but zone sizes correlated with MICs for gram-negative organisms (Fig. 2). Because of its in vitro spectrum of activity, LY163892 has the potential for clinical use in upper and lower respiratory tract infections, skin and skin structure infections, and urinary tract infections caused by susceptible bacteria. Its greater stability in comparison with cefaclor is an advantage.
This study was supported by a grant from Eli Lilly & Co.
LITERATURE CITED 1. Barry, A. L. 1986. The antimicrobic susceptibility test: principles and practices, p. 95. Lea & Febiger, Philadelphia. 2. Jones, R. N. (ed.). 1984. Performance standards for antimicrobial disk susceptibility tests, 3rd ed. National Committee for Clinical Laboratory Standards. Viltanova. Pa.

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