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Title page
Abstract
국문초록
CONTENTS
I. Introduction 13
II. Materials and Methods 17
1. Material and reagents 17
2. Bacterial strains 17
3. Animals 18
4. Drug susceptibility test 18
5. DNA analysis of quinolone-resistant S. pneumoniae with defined mutation in the QRDR 19
6. Time-kill studies 20
7. In vivo studies 21
8. The study of resistance mechanism 22
III. Results 24
1. Antibacterial activities of DW-224a 24
1.1 Susceptibility test against quinolone-susceptible strains 24
1.2 Susceptibility test against quinolone-resistant strains 27
2. DNA analysis of quinolone-resistant strains with defined mutation in QRDR 28
3. Time-kill studies 30
4. In vivo studies of DW-224a 33
4.1 Systemic infection model 33
4.2 Pneumococcal pneumonia model 34
5. The study of resistance mechanism of DW-224a 35
5.1 S. pneumoniae mutants selected by stepwise-mutation for DW-224a 35
5.2 Susceptibility test for other fluoroquinolones and analysis of point mutations in QRDR 37
IV. Discussion 40
V. References 43
Table 1. Primer sequences for amplification of QRDR 19
Table 2. The MICs (μg/ml) of tested organism against Ninety-four quinolone susceptible strains 25
Table 3. The MICs (μg/ml) of tested organism against twenty-three quinolone resistant strains a 27
Table 4. The MICs of quinolone for 23 strains with defined mutations in the QRDR a 29
Table 5. In vivo efficacies of quinolones on systemic infection model in mice 33
Table 6. Mutation frequencies of S. pneumoniae c9211 and its derivatives to DW-224a 36
Table 7. Fluoroquinolone susceptiblities and mutations identified in the QRDRs in DW-224a-resistant S. pneumoniae strains 39
Figure 1. Chemical structure of DW-224a 16
Figure 2. Bactericidal activity of DW-224a against S. pneumoniae 18 (PSSP). ■, 1/4× MIC; ●, 1/2× MIC; ▲, 1× MIC; ○, 2× MIC; □, 4× MIC. 31
Figure 3. Bactericidal activity of DW-224a against S. pneumoniae c9211 (PRSP & QSSP). ■, 1/4× MIC; ●, 1/2× MIC; ▲, 1× MIC; ○, 2× MIC; □, 4× MIC. 31
Figure 4. Bactericidal activity of DW-224a against S. pneumoniae 17 (PRSP & QRSP). ■, 1/4× MIC; ●, 1/2× MIC; ▲, 1× MIC; ○, 2× MIC; □, 4× MIC. 32
Figure 5. Therapeutic efficacies of DW-224a, ciprofloxacin, and gemifloxacin in a mouse model of pneumoniae due to S. pneumoniae ATCC 6305 34
Figure 6. Selection of S.pneumoniae c9211 mutants to DW-224a. The numbers outside the boxes show the concentrations of DW-224a used in each selection step. 36
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