Transcript No Slide Title

BMS-284756 (T-3811ME) Susceptibility Test Comparisons and Development for
More Than 30 Bacterial Species Using 3,328 Recent Clinical Isolates
Poster #717
8
2
6.7
10
6
10
33.3a 20.0a 33.3a
2
6.7a
0
0.0a
0
0.0
Gram-negative anaerobes (121)
%
2
1.6
43
49
b
35.5 40.5b
6
5.0b
2
1.7b
1
0.8
Gram-positive anaerobes (76)
%
18
14.9
12
30
31
c
c
15.8 39.5 40.8c
2
2.6
1
1.3c
0
0.0c
2
y
=
1
4
.
6
0
.
3
3
x
2
2
1
3
1
0
9
3
3
3
2
r
=
0
.
9
1
1
1
5
5
1
4
3
5
2
0
.
5
1
1
0
.
2
5
1
1
0
.
1
2
1
1
6
1
4
1
1
0
2
3
1
6
2
5
1
5
3
1
9
2
6
5





Acinetobacter (43)
P. aeruginosa (129)
Stenotrophomonas/
Burkholderia spp. (97)
TOTAL (269)
4
0
0
0
0
Etest MIC/reference MIC
2
1
0.5
0.25
5
6
4
15
4
38
31
73
0
33
29
62
0
1
12
13
FIGURE 1: Chemical Structure of BMS-284756
4
1
0
1
5
2
0
2
5
3
0 
3
5
on-scale results for both tests were used to assess essential agreement
(168 organism tests).
bEssential agreement  one log dilution was 89.3% (97.0%  two log dilutions steps).
2
2
0
.
2
5
63
0
.
1
2
1
58
26
7
0
.
0
6
2
2
8
9
8
3
9
1
1
25
96
0
.
0
3


0
.
0
1
6
0
.
0
3
2
0
.
0
6
4
0
.
1
2
5
0
.
2
5
0
.
5
1
2
4
8
1
6
3
2
3
2
3
1
4
4
2
1
 Overall, excellent inter-method, (Etest, BMD, DD, AD) correlation coefficients
1
1
2
7
4
2
3
1
A
l
l
E
n
t
e
r
o
c
o
c
c
i
(
n
=
9
9
)
2
O
y
=
1
4
.
2
0
.
2
9
x
2
1
1
3
4
1
1
r
=
0
.
9
3
1
CO 2H
1
0
.
5
6
2
2
2
1
1
0
.
2
5
1
4
3
4
2
2
2
0
.
1
2
2
2
2
1
N
HN
5
O
1
0
1
5
2
0
2
5
3
0
3
5
B
M
S
2
8
4
7
5
6
5

g
D
i
s
k
Z
o
n
e
D
i
a
m
e
t
e
r
(
m
m
)
REFERENCES
S
.
p
n
e
u
m
o
n
i
a
e
(
n
=
3
2
7
)
4
y
=
1
1
.
1
0
.
2
0
x
1
2
4
8
2
5
1
3
2

4
4
4
7
7
2
2
1
E
n
t
e
r
i
c
b
a
c
i
l
l
i
(
n
=
6
5
6
)
y
=
1
4
.
5
0
.
3
4
x
r
=
0
.
9
4
2
1
2
9
0
7
1
2
0
.
5
1
2
7
6
3
6
4
5
2
4
2
1
0
.
2
5
1
4
1
4
6
3
9
4
1
2
3
1
1
0
.
1
2
1
1
2
1
6
3
3
2
4
1
7
5
3
5
2
2
1
1
5
1
1
2
2
3
1
9
8
2
9
4
8
1
1
0
.
0
3

1
1
4
1
2
3
3
0
2
2
1
7
0
2
9
9
5
2
5
1
1
1
1
2
3
0
.
5
2
0
.
2
5
1
0
.
1
2
1
5
3
3
3
4
5
3
1
0
.
0
6
2
1
2
2
2
0
2
2
4
3
4
9
3
8
1
1
6
4
0
3

0
.
0
3
1
1
1
2
1
2
1
1
1
1
7
6
4
1
5
1
0
1
5
2
0
2
5
3
0
3
5
B
M
S
2
8
4
7
5
6
5

g
D
i
s
k
Z
o
n
e
D
i
a
m
e
t
e
r
(
m
m
)
1
0 1
5 2
0 2
5 3
0
3
5 4
0
B
M
S
2
8
4
7
5
6
5

g
D
i
s
k
Z
o
n
e
D
i
a
m
e
t
e
r
(
m
m
)
FIGURE 3: Scattergrams comparing the BMS-284756 MIC with the zone of inhibition around a 5-g disk when
testing P. aeruginosa, (124 strains). Vertical broken lines indicate preliminary zone diameter breakpoint criteria
correlating to the proposed  4 g/ml MIC indicating susceptibility.
FIGURE 7: Scattergram comparing the reference agar dilution BMS-284756 MICs and zone diameters
around 5-g disks (137 strains of N. gonorrhoeae; r=0.78)
4
.
0
1
1
1
3
3
1
2
1
P
.
a
e
r
u
g
i
n
o
s
a
(
n
=
1
2
4
)
2
1
1
3
4
4
1
1
1
1
1
.
0
0
.
5
0
.
5
r
=
0
.
9
5
1
1
5
3
4
1
2
0
.
2
5
0
.
1
2
1
1
0
.
0
6
5
1
5
2
0
2
5
3
0 
3
5
B
M
S
2
8
4
7
5
6
5

g
D
i
s
k
Z
o
n
e
D
i
a
m
e
t
e
r
(
m
m
)
4
0
2
0
.
2
5
4
1
1
2
1
1
2
1
1
1
1
1
11
1
0
.
0
6
N
.
g
o
n
o
r
r
h
o
e
a
e
(
n
=
1
3
7
)
2
1
1
1
3
2
1
1
1
1
1
1
y
=
1
2
.
5
0
.
1
8
x
0
.
0
3
2. Fung-Tomc JC, Minassian B, Kolek B, Huczko E, Aleksunes L, Stickle T, Tasho
T, Gradelski E, Valera L, Bonner DP. (2000a). Antibacterial spectrum of a novel
des-fluoro(6)quinolone, BMS-284756. Antimicrob Agents Chemother 44:33513356.
3. Hoellman DB, Kelly LM, Jacobs MR, Appelbaum PC. (2001). Comparative
antianaerobic activity of BMS-284756. Antimicrobial Agents Chemother 45,
589-592.
4. National Committee for Clinical Laboratory Standards (NCCLS). (2000).
Methods for dilution antimicrobial susceptibility tests for bacteria that grow
aerobically. Document M7-A5. Wayne, PA:NCCLS.
1
1
2
3
1
1
1
2
0
.
1
2
r
=
0
.
7
8
1
2
2
2
1
2
5
2
5
3
3
3
2
1
0
.
0
1
6
2
1
1
4
5
4
4
1
3
3
3
4
1
1
1
0
.
0
0
8
1
1
2
3
1
2
0
.
0
0
4
1
0
1
y
=
1
3
.
3
0
.
2
0
x
1
2
8
2
4
7
2
1
1
1. Biedenbach DJ, Croco MAT, Barrett TJ, Jones RN. Comparative in vitro activity
of gatifloxacin against Stenotrophomonas maltophilia and Burkholderia species
isolates including evaluation of disk diffusion and Etest methods.
Eur J Clin Microbiol Infect Dis 1999; 18:428-431.
5. National Committee for Clinical Laboratory Standards (NCCLS). (2000).
Performance standards for antimicrobial disk susceptibility tests, seventh edition.
Approved standard, M2-A7. Wayne, PA:NCCLS.
2
.
0
4
accurate susceptibility testing results for BMS-284756 when testing a wide
variety of bacteria.

4
r
=
0
.
6
6
2
 The Etest, AD, and disk diffusion testing methods can be used to provide
FIGURE 6: Scattergram comparison BMS-284756 MIC and zone diameter (5-g disk) results for 327
S. pneumoniae.
FIGURE 2: Scattergram comparing reference BMS-284756 MIC results to the zone diameters around 5-g disks.
Broken vertical line represents interpretive breakpoint suggested for Enterobacteriaceae (susceptible at  15 mm,
resistant at  11 mm and a  4 g/ml MIC breakpoint).
1
1
6
5
3
2
1
1
 The 5-g disk concentration appears to be appropriate for MIC breakpoints up

0
.
0
3
CHF2
4
(generally r  0.90) were observed for enteric bacilli, pseudomonads,
Acinetobacter, staphylococci, enterococci, Campylobacter, and Gram-negative
and -positive anaerobes.
to  2 g/ml. A larger disk content of BMS-284756 may be required for a higher
breakpoint ( 4 g/ml).
0
.
0
6

0
.
0
3
aOnly
3
CONCLUSIONS
0.12
5
5
0
.
5
B
M
S
2
8
4
7
5
6
E
t
e
s
t
M
I
C
(

g
/
m
l
)

4
2
2
2
1
1
(
a
)
4
0
0
254
FIGURE 5: Scattergram comparing the reference broth dilution BMS-284756 MICs and zone diameters
around 5-g disks for enterococci (99 strains; r = 0.93). Suggested breakpoints are shown as solid
vertical and horizontal lines. More conservative criteria are noted as broken lines.
1

4
0
.
0
6
TABLE 1: In vitro susceptibility of 269 non-fermentative
Gram-negative blood stream infection isolates from the
SENTRY Program (2000). Comparison of BMS-284756
Etest versus BMD MICs
1
B
M
S
2
8
4
7
5
6
5

g
D
i
s
k
Z
o
n
e
D
i
a
m
e
t
e
r
(
m
m
)
BMS284756IC( g/ml)

1
1
1
2
4
5
7
1
4
3
3
4
7

0
.
0
3
BMS284756IC( g/ml)

2
1
2
1
0
.
0
6
0
0.0
1
r
=
0
.
7
4
1
1
1
2
3
4
1
1
2
1
C. jejuni (30)
%
y
=
1
.
3
+
0
.
5
4
x
4
1
1
1
1
A
l
l
S
t
a
p
h
y
l
o
c
o
c
c
i
(
n
=
3
0
0
)
 two log2 dilutions was 93.3% (60.0% at  one log2 dilution).
bPercentage  two log dilutions was 97.6% (60.4% at  one log dilution).
2
2
cPercentage  two log dilutions was 97.4% (81.6% at  one log dilution).
2
2
inhibition an excellent correlation was achieved (r=0.94).
The Etest versus BMD essential agreement was 97.7% within  one log2 dilution and
99.8% when  two log2 dilutions were compared for enteric bacilli.
Inter-method comparison results for BMS-284756 DD versus reference MIC was
r=0.95.
The summary of the non-fermentor strains testing BMS-284756 Etest versus reference
MIC showed an essential agreement  one log2 dilution of 89.3% (97.0% for  two log2
dilutions).
When comparing BMS-284756 MIC to disk zone diameters for staphylococci (both
oxacillin-susceptible and -resistant included) an excellent linear correlation developed,
r=0.91.
The correlation of r=0.93 was achieved when comparing BMS-284756 BMD MICs to
disk zone diameters for all enterococci tested.
Excellent linear correlation was observed for Etest versus reference BMD results
(r=0.74) with 99.7% of results within  one log2 dilution for the streptococci group
including S. pneumoniae, viridans group streptococci, and -haemolytic streptococci.
Evaluations and comparisons of BMS-284756 MICs and disk zone diameters showed
that only a susceptible breakpoint was necessary and quantitative or categorical
agreement was 79.5% and 100.0%, respectively when testing H. influenzae.
Testing penicillin-susceptible or -resistant and ciprofloxacin-resistant strains of
gonococci produced an acceptable correlation (r=0.78) between BMS-284756 MICs
and disk zone diameter results.
Inter-method comparisons of BMS-284756 Etest MICs and AD MICs were 93.3%,
97.6%, and 97.4% for results with C. jejuni, Gram-negative and -positive anaerobes,
respectively when using criteria within  two log2 dilutions.
Organism (no. tested)
Isolates (3,328 strains) were selected from various clinical organism collections at
the University of Iowa College of Medicine (Iowa City, IA), JMI Laboratories, and
the SENTRY Antimicrobial Surveillance Program (1997-2001). These isolates
belong to over 30 different genus groups. Most isolates were stored at -70C or
below in serum or lysed blood, or at room temperature in distilled water
(Pseudomonas spp.).
Organism (no. tested)
4
BMS284756IC( g/ml)

INTRODUCTION
MATERIALS AND METHODS
A
l
l
S
t
r
e
p
t
o
c
o
c
c
i
(
n
=
6
6
8
)
3
 When comparing enteric bacilli reference BMD MIC results to the zone diameters of

Fluoroquinolone therapy has been steadily increasing world wide over the last 12 14 years due to a wide spectrum of activity, low toxicity, oral or parenteral dosing,
and good potency enhanced by very favorable pharmakinetics. Fluoroquinolone
therapy is often prescribed empirically or directed by susceptibility test results for
multitudes of infections including nosocomial and community-acquired respiratory
tract therapy (sinusitis, pneumonia and chronic bronchitis).
BMS-284756 is a novel des-F(6)-quinolone (formerly T-3811ME) with a broad
spectrum of activity. While it may be no more active than ciprofloxacin or
levofloxacin against many Gram-negative bacteria, it is consistently more active
against the Gram-positive bacteria. Animal model studies show BMS-284756 has
greater bioavailability than ciprofloxacin and is less toxic than levofloxacin.
The study is designed to evaluate the in vitro susceptibility testing criteria of BMS284756 by comparing over 30 species of Gram-positive and -negative bacteria by
reference broth microdilution (BMD), agar dilution, and disk diffusion methods.
Comparisons between reference and Etest MICs were also performed to access
accuracy of the agar stable gradient methodology. The proposed breakpoint of
 4 g/ml was used for some MIC comparisons.
Occurrances at Etest/AD MIC ratio:
 0.125 0.25 0.5
1
2
4
FIGURE 8: Comparison of BMS-284756 MIC results obtained with the Etest and the reference NCCLS
method for all streptococci tested (668 strains). Only two of 674 strains varied beyond the  one log2
dilution range (99.7% agreement between methods).

4

4
aPercentage
RESULTS
FIGURE 4: Scattergram comparing the reference broth microdilution BMS-284756 MICs and zone
diameters around 5-g disks for staphylococci (300 strains; r = 0.91). Suggested breakpoints are
shown as solid vertical and horizontal lines. More conservative criteria separating two distinct organism
populations are noted as broken lines.
BMS284756IC( g/ml)
Testing was performed according to reference National Committee for Clinical
Laboratory Standards (NCCLS) recommended procedures for broth microdilution
(BMD), agar dilution (AD) and disk diffusion (DD) for each specific species. The Etest
(AB BIODISK, Solna, Sweden) was performed according to the method described in
the manufacturer’s product package insert.
BMS-284756 laboratory standard powder was supplied by Bristol-Myers Squibb
(Princeton, NJ) and the comparison drugs by their domestic manufacturers.
The BMS-284756 5-g disks and Etest strips were made by BD Microbiology Systems
(Cockeysville, MD) and AB BIODISK, respectively.
Results of AD, Etest and BMD MICs when available were compared to establish
essential agreement between the results of these quantitative methods. MICs and DD
zone diamters were analyzed by regression analysis and error-rate bounding to
determine correlation between results.
TABLE 2: Comparison of Etest and agar dilution (AD)
test results for BMS284756 versus strains of C. jejuni,
Gram-negative anaerobes and Gram-positive anaerobes
BMS284756IC( g/ml)
MATERIALS AND METHODS – CON’T
BMS284756IC(mg/L)
Background: BMS-284756 (BMS) is a novel des-F(6)-quinolone with activity
against many Enterobacteriaceae, non-fermentative Gram-negative bacilli,
staphylococci, streptococci, and fastidious or anaerobic species. In this report we
compare BMS results between various in vitro susceptibility testing methods and
propose breakpoint criteria.
Methods: BMS activity was evaluated by NCCLS broth microdilution (BMD), agar
dilution (AD), 5-g disk diffusion (DD) and Etest methods against 3,328 bacterial
strains from various organism collections including the SENTRY Program
(1997-2000).
Results: The correlation coefficients (r) between BMD and Etest MIC values for
the 11 species of Enterobacteriaceae, P. aeruginosa, and Acinetobacter spp.
ranged 0.92-0.98. BMD vs. DD results were also excellent (r=0.94-0.99). The
nonfermenter group Burkholderia/Stenotrophomonas spp. had a lower (r) for both
BMD vs. Etest and BMD vs. DD (0.81-0.84). Staphylococci and enterococci were
only compared for BMD vs. DD with an overall (r) of 0.94 (400 strains).
Streptococci had a reduced correlation for both BMD vs. Etest and vs. DD,
ranging from 0.44 to 0.80, but the Etest MIC value  1 log2 dilution was 99.4%.
The BMD vs. Etest MIC (r) was consistently better than BMD vs. DD. H.
influenzae (292 strains) had higher Etest BMS MICs with low (r) of 0.12 and 0.29,
yet perfect categorical agreement. AD results were compared to DD and Etest
values (r=0.78-87) against N. gonorrhoeae (137 strains). Etest results were
consistently 4-fold lower than the AD MICs. C. jejuni and anaerobes when tested
by AD and Etest showed 93-98% of results  2 log2 dilutions.
Conclusions: The inter-method correlation (r) for BMS results were greater than
0.90 for nearly all common species evaluated and without serious categorical
error using a proposed BMS breakpoint of 4 g/ml. Both Etest and DD
approximations were considered to be highly accurate alternatives for testing BMS
potency in clinical laboratories.
P.R. Rhomberg, D.J. Biedenbach, R.N. Jones.
The JONES Group/JMI Laboratories, North Liberty, IA
BMS284756IC( g/ml)
ABSTRACT
Ronald N. Jones, M.D.
The JONES Group / JMI Laboratories
345 Beaver Kreek Centre, Suite A, North Liberty, Iowa 52317
Phone: 319-665-3370 Fax: 319-665-3371
[email protected]
1
1
1
1
5
1
0
1
5
2
0
2
5
3
0
3
5
4
0
4
5
5
0
5
5

6
0
6
5
B
M
S
2
8
4
7
5
6
5

g
D
i
s
k
Z
o
n
e
D
i
a
m
e
t
e
r
(
m
m
)
6. National Committee for Clinical Laboratory Standards (NCCLS). (2001).
Development of in vitro susceptibility testing criteria and quality-control
parameters. Approved guideline M23-A2. Wayne, PA:NCCLS.
7. Takahata M, Mitsuyama J, Yamashiro Y, Yonezawa M, Araki H. Todo Y,
Minami S, Watanabe Y, Narita H. (1999). In vitro and in vivo antimicrobial
activities of T-3811ME, a novel des-F(6)-quinolone. Antimicrob Agents
Chemother 43:1077-1084.
A156-23