舒巴坦联合美罗培南不同给药方案抑制鲍曼不动杆菌耐药突变的研究

发布时间：2018-11-10 16:57

【摘要】：目的：在体外PK/PD模型中,探讨舒巴坦联合美罗培南的不同给药方案对鲍曼不动杆菌耐药突变的抑制作用。方法：纸片扩散法测定抗菌药物单药对鲍曼不动杆菌45791的MIC值；微量肉汤稀释法和琼脂法测定美罗培南和舒巴坦单药及联合时对该菌的MIC、MPC值和两药的相互作用。建立体外药代动力学/药效学(PK/PD)模型,模拟2种美罗培南单药给药方案(1.030min Q8h,1.03h Q8h);6种舒巴坦单药给药方案,即1.030min Q8h、2.030min Q8h、3.030min Q8h及1.03h Q8h、2.03h Q8h、3.03h Q8h;6种联合方案,即美罗培南1.030min Q8h分别与舒巴坦1.030min Q8h、2.030min Q8h、3.030min Q8h,美罗培南1.03h Q8h分别与舒巴坦1.03h Q8h、2.03h Q8h、3.03h Q8h。各方案均持续给药7天,在相应的时间点测定模型中总细菌数、耐药突变菌数,并用高效液相色谱法(HPLC)测定美罗培南的浓度,用高效液相色谱串联质谱法(HPLC-MS/MS)测定肉汤中舒巴坦的浓度。结果：美罗培南对该菌的MIC为2μg·mL-1,MPC为28.μg·mL-1,舒巴坦的MIC为128μg·mL-1,MPC为128μg·mL-1,该菌对青霉素类、喹诺酮类、氨基糖苷类、头孢菌素类、含β内酰胺酶的复合制剂类耐药。棋盘法表明,联合的MIC显示有协同作用,联合后美罗培南和舒巴坦的MPC值各降至8ug/mL和32ug/mL。6种舒巴坦单药方案中,168h内细菌总量均未见减少。美罗培南1.030min Q8h及1.03hQ8h单药方案中,总细菌量在72h及24h内持续下降,下降约103cfu/mL,之后分别在144h和96h内维持基本不变,最后稳定生长,最终浓度超过106cfu/mL;耐药突变菌于72h富集生长。6种联合方案均不能防止耐药菌产生,美罗培南1.030min Q8h与舒巴坦1.030min Q8h、2.030min Q8h、3.030minQ8h的联合方案中,各联方案之间无差异；总细菌数持续下降,下降约105cfu/mL,耐药突变菌于144h富集生长。美罗培南1.03h Q8h与舒巴坦1.03h Q8h、2.03h Q8h、3.03h Q8h联合方案中,各联方案之间亦无差异；总细菌数持续下降,下降约106cfu/mL,未出现耐药菌富集生长。上述14种方案,在时间为168h时系统中的细菌均全部为耐药菌。结论：对于本研究的实验菌株,舒巴坦和美罗培南联合可以明显缩小美罗培南的突变选择窗口。舒巴坦与美罗培南联合后,可明显延缓耐药突变菌的富集速度。3小时静脉滴注的联合给药方案比30min静脉滴注的联合给药方案效果更明显,能在168小时内抑制耐药菌的富集。美罗培南与舒巴坦的联合中,增加舒巴坦的剂量不能提高杀菌效果和抑制耐药突变。
[Abstract]:Aim: to investigate the inhibitory effect of sulbactam combined with meropenem on resistant mutation of Acinetobacter baumannii in vitro PK/PD model. Methods: the MIC value of single antimicrobial drug against Acinetobacter baumannii 45791 was determined by disk diffusion method, and the MIC,MPC value and the interaction between the two drugs were determined by broth dilution method and Agar method. In vitro pharmacokinetics / pharmacodynamics (PK/PD) model was established to simulate two meropenem single drug administration regimens (1.030min Q8h 1.03h Q8h). Six sulbactam single drug administration regimens, namely 1.030min Q8H 2.030min, Q8h 3.030min Q8h and 1.03h Q8hU 2.03h Q8hU 3.03hQ8h; Six combined schemes, meropenem 1.030min Q8h and sulbactam 1.030min Q8hU 2.030min Q8h, meropenem 1.03h Q8h and sulbactam 1.03h Q8h respectively and sulbactam 1.030min Q8hU 2.030min Q8hU 3.030min Q8h respectively. The total number of bacteria and the number of resistant mutant bacteria in the model were determined at the corresponding time points. The concentration of meropenem was determined by high performance liquid chromatography (HPLC) (HPLC). The concentration of sulbactam in broth was determined by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Results: the MIC of meropenem to this strain was 2 渭 g mL-1,MPC = 28. 0 渭 g mL-1, sulbactam, and the MIC of meropenem was 128 渭 g mL-1,MPC and 128 渭 g mL-1,. The bacteria had penicillin, quinolones, aminoglycosides, cephalosporins, penicillin, quinolones, aminoglycosides and cephalosporins. Complex preparations containing 尾-lactamases were resistant to drugs. Chessboard method showed that the combined MIC showed synergistic effect, and the MPC values of meropenem and sulbactam decreased respectively to 8ug/mL and 32ug/mL.6 sulbactam monopharmaceuticals, and the total amount of bacteria did not decrease within 168h. In meropenem 1.030min Q8h and 1.03hQ8h single drug regimen, the total bacterial count decreased continuously in 72 h and 24 h, decreased about 103 cfur / mL, then remained basically unchanged for 144 h and 96 h, and finally grew stably, and the final concentration was over 106 cfur / mL; The drug resistant mutant bacteria were enriched and grown at 72 h. All of the six combination protocols could not prevent the production of resistant bacteria, but there was no difference between the two combinations of meropenem 1.030min Q8h and sulbactam 1.030min Q8h 2.030min Q8hU 3.030minQ8h. The total bacterial count continued to decrease, about 105 cfur / mL, and the drug-resistant mutant bacteria enriched and grew at 144h. In the combination regimen of meropenem 1.03h Q8h and sulbactam 1.03h Q8hU 2.03h Q8hU 3.03h Q8h, there was no difference between the two schemes, and the total bacterial count decreased by 106cfu-mL. there was no enrichment growth of drug-resistant bacteria. All the bacteria in the system were drug-resistant when the time was 168h. Conclusion: the combination of sulbactam and meropenem can significantly reduce the mutation window of meropenem. After combination of sulbactam and meropenem, the enrichment rate of resistant mutant bacteria was significantly delayed. The combined administration regimen of 3 hours intravenous drip was more effective than that of 30min intravenous drip regimen, and could inhibit the enrichment of resistant bacteria within 168h. In the combination of meropenem and sulbactam, increasing the dosage of sulbactam did not improve bactericidal efficacy and inhibit drug-resistant mutation.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

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