CHEMOTHERAPY NOV Fig. 1 Imipenem (MK-0787) Enterobacter cloacae Enterobacter aero Morganella morganii Pseudo- Acinet ob acter Staphylococcus aur

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1 CHEMOTHERAPY NOV Fig. 1 Imipenem (MK-0787) Enterobacter cloacae Enterobacter aero Morganella morganii Pseudo- Acinet ob acter Staphylococcus aureus Streptococcus pyogenes Escherichia coli Klebsiella pneumoniae genes Serratia marcescens Proteus mi rabilis Proteus vulgaris Providencia calco aceticus Haemophilus influenzae Imipenem (MK-0787), ceftizoxime (CZX), latamoxef (LMOX), cefoperazone (CPZ), cefmetazole (CMZ), cefsulodin (CFS), ceftazidime (CAZ), piperacillin(pipc), sulbenicillin (SBPC)genta-

2 VOL. 33 S-4 CHEMOTHERAPY Table 1 Antibacterial spectrum of MK-0787, CPZ, CZX, LMOX and PIPC Gram-positive bacteria Inoculum size: 106cells/ml MIC (Đg/ml) *Sensitivity test agar supplement with 10% horse blood Table 2 Antibacterial spectrum of MK-0787, CPZ, CZX, LMOX and PIPC Gram-positive bacteria Inoculum size: 108 ells/ml MIC (Đg/ml) *Sensitivity test agar supplemented with 10% horse blood Streptococcus Corynebacterium diphtheriae H. influenzae

3 CHEMOTHERAPY NOV. 198B Table 3 Antibacterial spectrum of MK-0787, CPZ, CZX, LMOX and PIPC Gram-negative bacteria MIC (Đg/ml)

4 CHEMO THERAPY Table 4 Antibacterial spectrum of MK-0787, CPZ, CZX, LMOX and PIPC Gram-negative bacteria Inoculum size: 108 cells/mlmic *Gonococcus medi um (fig/m1)

5 CHEMOTHERAPY Table 5 Antibacterial spectrum of MK-0787, CPZ, CZX, LMOX and PIPC anaerobic bacteria Inoculum size: 108 cells/m1mic (peml) Table 6 Antibacterial spectrum of MK-0787, CPZ, CZX, LMOX and PIPC Anaerobic bacteria Inoculum size: 108 cells/mlmic (pg/m1)

6 CHEMO THERAPY Fig. 2 Sensitivity distribution of clinical isolates S. aureus (42 strains) Fig. 4 Sensitivity distribution of clinical isolates. E. coli (59 strains) Fig. 3 Sensitivity distribution of clinical isolates S. pyogenes (31 strains) Fig. 5 Sensitivity distribution of clinical isolates K. pneumoniae (59 strains)

7 CHEMO THERAPY Fig. 6 Sensitivity distribution of clinical isolates E. cloacae (30 strains) Fig. 8 Sensitivity distribution of clinical isolates S. marcescens (52 strains) Fig. 7 Sensitivity distribution of clinical isolates Fig. 9 Sensitivity distribution of clinical isolates E. aerogenes (34 strains) P.mirabilis (32 strains)

8 CHEMO THERAPY Fig.10 Sensitivity distribution of clinical isolates Fig. 12 Sensitvity distribution of clinical isolates P.ulgaris (32 strains) M. morganii (25 strains) Fig.11 Sensitivity distribution of clinical isolates Fig. 13 Sensitivity distribution of clinical isolates P.rett geri (15 strains) P.aeruginosa (40 strains)

9 CHEMO THERAPY Fig.14 Sensitivity distribution of clinical isolates Fig.16 Sensitivity distribution of clinical A. calcoaceticus (44 strains) GMr, SBPCr P.aeruginosa (37 strains) Fig.15 Sensitivity distribution of clinical isolates Fig.17 Sensitivity distribution of clinical H. influenzae (48 strains) CFSr P.aeruginosa (35 strains)

10 CHEMO THERAPY Table 7 Antibacterial activity of MK-0787 *80% Inhibitory concentrations of MK-0787, CZX, CMZ and LMOX Table 8 Antibacterial activity of MK-0787 on GM, SBPC and CFS resistant P. aeruginosa

11 CHEMO THERAPY Table 9 Influence of inoculum size on the antibacterial activity of MK-0787 MIC: ml Table 10 Influence of medium ph on the antibacterial activity of MK-0787 Inoculum size : 10 cells/m1 MIC: ml

12 CHEMOTHERAPY Table 11 Influence of horse serum on the antibacterial activity of MK-0787 Inoculum size: MIC: pg/m1 106 cells/1-n] Fig. 18 Effect of MK-0787 on the viability of S. aureus 209-P JC Fig. 20 Effect of MK-0787 on the viability of K. pneumoniae KC-1 Fig. 19 Effect of MK-0787 on the viability of E. coli K-12 Fig. 21 Effect of MK-0787 on the viability of S. marcescens T-55

13 CHEMOTHERAPY 86 Fig. 22 Effect of MK-0787 P. aeruginosa E-2 on the viability NOV μg/mlに of あ り,CFSの6.25μg/ml,CPZの12.5 μg/ml,gmの3.13お よ び50μg/ml,SBPCの12.5 お よ び100μg/mlよ 値 をTable8に 6.25μg/mlや り も優 れ て い た そ の 時 のMIC80 示 した 本 剤 は3.13μg/mlでCFSの 他 の比 較 薬 よ りも優 れ て いた 16)CFSrP. aeruginosaの 1983年 場合 に京 滋地 区 の 呼吸 器 感 染 症 患 者 よ り分離 され たP. aeruginosaの 中 で,CFS耐 性 の 結 果 をFig.17に 3.13μg/mlに 性35株 に 対 す る感受 示 し た 本 剤 の 感 受 性 の ピ ー クは あ り,GMは3.13μg/mi,CAZは025 μg/ml,cpzは25μg/ml,cfsは>100μg/mlで た MIC80値 あっ で み て も 本 剤 は3.13μg/mlで あ り,他 の 比較 薬 に比 べ て 明 らか に 優 れ て い た 3.抗 菌 力 に及 ぼ す 諸 因 子 の影 響 s209-p い た 14)H. influenzaeの Fig.15に iaekc-1お 場合 示 す よ う にMK-0787の 0.78μg/mlに 感 受 性 ピー ク値 は り劣 り,CMZの6.25μg/mlよ た MIC80値 の 比 較 で も,本 り優 れ て い 剤 は3.13μg/mlで あ り, CZXの0.05μg/mlやLMOXの0.2μg/mlよ り劣 っ て い た 15)GMr,SBPCrP. aeruginosaの Fig.16に Fig. 示 す よ うにMK-0787の 23 Phase-contrast よ びP. aeruginosae-2を 力 に 及 ぼ す 培 地pH,馬 場 合 感 受 性 ピー ク値 は micrographs of E. coli 9 11に pneumo-n 用 い て,抗 菌 血 清 添 加,接 CZX,LMOX,CPZ,PIPCと あ り,CZXの0.05μg/mlやLMOXの 0.1μg/mlよ JC, E.coliNIHJJC-2,K. S.aureu 種菌量の影響を 比 較 した 結 果 をTable 示 した 本 剤 を 含 む す べ て の 薬 剤 は,い 菌 株 に お い て も,培 地pH,馬 ず れの 血 清 添 加 に お い て,MIC の 変 動 は ほ と ん ど 認 め ら れ な か っ た ま た 本 剤 は,接 種 菌 量 に よ る 影 響 を,S.aureusやP. aeruginosaで はほと ん ど 受 け な か っ た が,E.coli,K.pneumoniaeで は菌量 の 増 加 に 伴 い,感 K-12 exposed to 受 性 は 若 干 低 下 し た 一 方 比 較 薬 とし MK-0787 for 3 hours at 37 Ž

14 VOL. Fig. Fig CHEMOTHERAPY S-4 Phase-contrast 25 Phase micrographs contrast micrographs of S. of P. inarcescens aeruginosa 87 T-55 exposed to MK-0787 for 3 hours at 37 Ž E-2 exposed to MK-0787 for 3 hours at 37 Ž

15 CHEMOTHERAPY 88 て 用 い たPIPC,CZX,LMOXで よ びP. aeruginosaに NOV.1985 Fig. 27 はK.pneumoniaeお お い て 接 種 菌 量 celles/ml Fluorography showing competition of MK-0787 for [14C]-PCG binding in P. aeruginosa E-2 で菌 量 に よ る影響 が み られ た 4.殺 菌作用 Fig.18に 示 した よ う に,Smrm5に 0787は107,108cells/ml接 対 してMK- 種 共 にMIC(0.013μg/ml) 以 上 の 濃 度 で 殺 菌 的 に 作 用 した ooliやk.pneumoniaeに Fig.20に 対 E. し て は,Fig.19お 示 した よ う にMK-0787の (い ず れ も0.78μg/ml)以 よび そ れ ぞ れ のMIC 上 の 濃 度 でdoseresponseの あ る殺 菌 作用 が み られ た marcescensの 場 合 は,Fig.21に S. 量 が107cells/mlの 示 した よ う に,菌 場 合,MIC(3.13μg/ml)以 で 殺 菌 作 用 が 見 ら れ た が,108cells/mlの 上 の 濃度 菌 量 で はを の 殺 菌 性 が 減 弱 す る傾 向 を 認 め た P.P. aeruginosaに MIC(1.56μg/ml)以 Smrm3の 対 して はFig.22に 示 した よ う に 上 の 濃 度 で 殺 菌 作 用 が み られ た が, 場 合 と 同 様doseresponseは あ ま り顕 著 で 溶 菌 象 も観 察 され た は な か った 5.位 対P.すaeruginosa る 形 態 変 化 の 結 果 をFig.25に 相 差 顕 微鏡 に よる形 態 観 察 coliに 対 す るMK-0787の し た 0.013お 示 した 0.1μg/mI作 示 い く様 子 が 観 察 され,0.39,0,78,1.56(MIC) 用 で はcontrolと 同 6.25μg/ml作 よ び0,025μg/ml作 様 に 増 殖 して い く様 子 が,0.05,0.1お よ び0,39μg/ml 6.PBPsに coliお 用 では 菌 体 の球 形 化 や 溶菌 像 を観 察 す る こ とが で きた 0787の S.marce scensに 対 す る 形 態 変 化 の 結 果 をFig.24に した 0.19,0.39,0.78μg/ml作 び12.5μg/ml作 Fig. 26 用 で はbulge状 示 用 で は菌 は 増 殖 しな が ら 球 形 化 して い く様 子 が 観 察 さ れ,3.13(MIC)お,3.13, の 細 胞 やspheroplast様 対 す る親 和 性 よ びP. aeruginosaのpbpsに E. 親 和 性 をFig.26お 対 す るMK- よ びFig.27に,そ シ ト メ ー タ ー 処 理 に よ り 得 た50%阻 12に よ の細 胞 や 球 形 化 そ して Fluorography showing competition of MK-0787 for [14C]-PCG binding in E. coli K-12 用 で はbulge状 同 様 に増 殖 して 構 造 そ して溶 菌像 が 観 察 され た 作 用 で は 菌 体 の 球 形 化 が 観 察 さ れ た ま た0.78(MIC) μg/ml作 用 で はcontrolと 形E. 態 変 化 をFig.23に 示 した coliに 3の して デ ン 害 濃 度 をTable 対 E. し てMK-0787はPBPsの2 順 に 良 好 な 親 和 性 を 示 し,そ 度 は,1a,1bs, れ ぞ れ の50%阻 く0.008,0.008,0.048,0.48で 害濃 あ っ た ま たP. に 対 して はPBPご52=1bs,1a,3の 良 好 な 親 和 性 を 示 し,そ の50%阻 <0.008=<0.008,0.0094,0 0105で III.総 新 しい β-lactam系 加vitro抗 aeruginosa 順に 害 濃度 は そ れ ぞ れ あ った 括 お よび 考 察 抗 生 剤 で あ るMK-0787に 関す る 菌 力 に つ い て 検 討 を 行 っ た 本 剤 は,グ ラム 陽 性 菌 お よ び グ ラ ム 陰 性 菌 に 幅 広 い 抗 菌 ス ペ ク トラム を 有 し,Bacteroidesに も 抗 菌 力 を 示 した グ ラム陽 性 菌 に は 比 較 薬 と して 用 い たCMZ,CTZ,LMOXよ りも 良 好 な 抗 菌 力 を 示 し た グ ラ ム 陰 性 菌 で はP. aeruginosa,a. calcoaceticusお よ びA.faecalisに は 比較薬 より も優 れ た 抗 菌 力 を 示 し た が,E.coli,K.pneumoniae, Prolmε 属 で は,比 較 薬 に 比 べ て 抗 菌 力 が 若 干 低 か った ま た,P. maltophiliaに はSAINOら7)の は 抗 菌 力 を 示 さ な か っ た これ 報 告 し て い る よ う に,P. maltophiliaが

16 VOL. 33 S-4 CHEMOTHERAPY Table 12 Competition of MK-0787 with 14C-labeled penicillin G for binding to PBPs in cytoplasmic membranes of E. coli K-12 and P. aeruginosa E-2 in vivo * Concentration as amolar ratio to 14C-labeled penicillin G required for 50% inhibition binding of labeled penicillin G 2) NASU, M.; J. P. MASKELL, R. J. WILLIAMS & J. D. WILLIAMS: In vitro activity of MK 0787 (N-formimidoyl thienamycin) and other beta-lactam compounds against Bacteroides spp. Antimicrob. Agents Chemother. 20: 433 `436, ) NEU, H. C. & P. LABTHAVIKUL: Comparative in vitro activity of N-formimidoyl thienamycin against gram-positive and gram-negative aerobic and anaerobic species and its p-lactamase stability. Antimicrob. Agents Chemother. 21: , 1982 Chemotherapy 27: 559 `560, ) SPRATT, B. G.; Distinct penicillin binding proteins involved in the division, elongation and shape of Escherichia coli K-12. Proc. Natl. Acad. Sci., U. S. A. 72: , ) SAINO, Y. ; F. KOBAYASHI, M. INOUE & S. MI- 1) KROPP, H.; J. G. SUNDELOP, J. S. KAHAN, F. M. KAHAN & J. BIRNBAUM: MK 0787 (N-formimidoyl thienamycin) : Evaluation of in vitro and in vivo activities. Antimicrob. Agents Chemother. 17: 993 `4000, 1980 TSUHASHI: Purification and properties of inducible penicillin Ĉ-lactamase isolated from Pseudomonas maltophilia. Antimicrob. Agents Chemother. 22 : , 1982 Chemotherapy 28 (S-7): 35-41, ) HASHIZUME, T.; F. ISHINO, J. NAKAGAWA, S. TAMAKI & M. MATSUHASHI: Studies on the mechanism of action of imipenem(n-formimidoyl thienamycin) in vitro: Binding to the penicillin-binding proteins (PBPs) in Escherichia coli and Pseudomonas aeruginosa and inhibition of enzyme activities due to the PBPs in E. coli. J. Antibiot. 37: 394 `400, 1984

17 CHEMOTHERAPY NOV IN VITRO ACTIVITY OF IMIPENEM, A NEW CARBAPENEM TAKESHI NISHINO, KAZUNORI NAKAMURA, MAYUMI TANAKA, TOSHIMI GOTOH MASAKO OHTSUKI and TERUO TANINO Department of Microbiology, Kyoto Pharmaceutical University The bacteriological evaluation of imipenem(mk-0787), a new S-lactam antibiotic, was studied. the following results were obtained. MK-0787 showed a broad antibacterial spectrum against gram-positive, gram-negative bacteria with the exception of P. maltophilia and Bacteroides. The antibacterial activity of MK-0787 against gram-positive bacteria was superior to that of cefinetazole, ceftizoxime and latamoxef. It inhibited gram-negative bacteria resistant to other agents. It did not show cross resistance for gentamicin or cefsulodin resistant P. aeruginosa. There was minimal effect of inoculum size of E. coli and K. pneumoniae, and medium ph or horse serum had no effect on activity. Time-kill curve confirmed a dose response bactericidal activity of MK In morphological observation of E. coli and P. aeruginosa by phase contrast microscope, exposure to MK-0787 resulted in the formation of spherical cells, bulge forms and spheroplast-like structures. MK-0787 showed a high affinity for PBPs 2 of E. coli and P. aeruginosa.

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