头孢喹肟对Ⅱ型猪链球菌在感染小鼠体内的药动药效模型研究
发布时间：2019-06-03 17:38
【摘要】：猪链球菌是人畜共患菌,给养猪业及公共卫生安全构成了严重的威胁。其中Ⅱ型猪链球菌是流行最广,致病性最强,危害最大的血清型。β-内酰胺类药物是治疗Ⅱ型猪链球菌感染首选药物,头孢喹肟对Ⅱ型猪链球菌极其敏感,是治疗Ⅱ型猪链球菌感染理想选择。目前有关Ⅱ型猪链球菌药动药效研究还未见报道,因此无法为治疗Ⅱ型猪链球菌感染提供科学依据。为此本文开展头孢喹肟对Ⅱ型猪链球菌的体外药效和体内药动药效同步模型的相关研究。本文主要通过体内头孢喹肟对Ⅱ型猪链球菌PK/PD参数、抗菌后效应和菌量效应研究,得到的数据通过蒙特克罗模拟推荐临床猪感染Ⅱ型猪链球菌的治疗方案。1.头孢喹肟对Ⅱ型猪链球菌的体外抗菌活性测定头孢喹肟对不同初始菌量(正常菌量:106log10CFU和高菌量:108log10CFU)的MIC和体外杀菌曲线,结果显示无论正常菌量还是高菌量MIC值差异不显著。体外杀菌曲线显示高菌量抑制头孢喹肟的杀菌效果,呈现菌量效应。对防突变浓度的测定结果显示,防突变浓度为0.12μg/m L,耐药选择突变窗为0.03~0.12μg/m L,选择指数为4,比值较低。体外实验表明:菌量变化对MIC影响不明显,但影响体外杀菌效果;选择突变窗窄表明不容易选择出耐药突变株。2.头孢喹肟在感染小鼠体内的药代动力学研究建立免疫缺陷小鼠大腿局部感染模型,按4倍增加的剂量治疗,于给药前和给药后从小鼠眼框静脉丛穿刺间隔采血,利用LC-MS/MS方法测定药物浓度,Win Nonlin5.2.1药动学软件对血药浓度-时间数据进行处理。结果显示:头孢喹肟在感染小鼠体内的药动学符合一级吸收一室模型。头孢喹肟的峰浓度为1.96~606.71μg/m L;药时曲线下面积为1.58~551.4μg.h/m L;达峰时间为0.25~0.28 h;消除半衰期为0.32~0.38 h,为后续药效学奠定基础。3.头孢喹肟对Ⅱ型猪链球菌的抗菌后效应为了减少杂菌干扰,本文优化和制定选择性培养基:萘啶酸:多粘菌素B(60:30,μg/m L:μg/m L)用于体内药效研究。应用免疫缺陷小鼠大腿局部感染模型,进行体内抗菌后效应(PAE)实验。结果显示:2.5mg/kg和40 mg/kg产生的PAE分别为2.45 h和8.55 h,具有浓度依赖性。结果表明,头孢喹肟对Ⅱ型猪链球菌可产生长PAE,相对于产生短PAE的细菌,治疗Ⅱ型猪链球菌时可相对延长给药间隔,此为首次在体内揭示β-内酰胺类药物对Ⅱ型猪链球菌可产生长的PAE。4.小鼠大腿局部感染模型的药动/药效同步模型研究应用免疫缺陷小鼠大腿局部感染正常菌量和高菌量制作感染模型,进行药动药效学研究。通过剂量分配法评价最相关的PK/PD指数、指数范围并考察是否存在菌量效应。结果显示:头孢喹肟对Ⅱ型猪链球菌在两种菌量下体内抗菌活性与PK/PD指数(%?TMIC)的相关性最强;指数范围:两个菌量达到静态效应所需的%?TMIC没有差异而达到1-log杀菌效时,感染高菌量所需的%?TMIC比正常菌量高1.2倍;根据剂量比例,高菌量达到静态效应和1-log杀菌效应所需的给药剂量与正常菌量的比例为1.59和4.47倍,结果表明,头孢喹肟药效受感染菌量的影响。接着结合头孢喹肟在猪体内的药动数据,通过蒙特卡罗分析,推出治疗临床上猪感染Ⅱ型猪链球菌的推荐给药剂量为:当MIC≤0.12μg/m L时,推荐给药剂量为2mg/kg/24h i.m.,当MIC=0.24μg/m L时,推荐给药剂量为2mg/kg/12h i.m。以上结果表明,头孢喹肟用于临床治疗Ⅱ型猪链球菌感染时,需要根据MIC值和感染程度调整给药方案。5.小鼠败血症模型药动/药效同步模型研究由于Ⅱ型猪链球菌引起猪和人类的感染主要为败血症,因此建立小鼠败血症模型进一步评价药效。给予4倍增加的5个药物剂量,间隔6h给药,24h治疗,以生存率作为判断标准,结果显示:药物半数有效量为2.87mg/kg,%TMIC为22.5%~42.5%,表明头孢喹肟在感染败血症的小鼠模型中也能达到很好的治疗效果。本研究利用体外药效和小鼠体内PK/PD模型,研究头孢喹肟对Ⅱ型猪链球菌的抗菌活性,确定了与抗菌效应最相关的PK/PD参数,计算不同感染菌量达到不同抗菌效应所需的PK/PD参数值,结合头孢喹肟在猪体内的药动数据通过蒙特卡罗分析推荐临床治疗猪感染Ⅱ型猪链球菌最佳治疗方案,为临床经验治疗提供科学依据。
[Abstract]:Streptococcus suis is a common animal, and poses a serious threat to the pig industry and public health. The second type of Streptococcus suis is the most popular and most highly pathogenic serotype. The present invention relates to a method for treating type II streptococcus suis infection, and is an ideal choice for treating type II streptococcus suis infection. At present, the study on the drug effect of type II streptococcus suis has not been reported, so it is not possible to provide scientific basis for the treatment of type II streptococcus suis infection. In this paper, the effect of cefixime on the in vitro and in vivo drug effect of Streptococcus suis is studied in this paper. The results of the study on the PK/ PD parameters, the post-antibiotic effect and the amount of the bacteria in the second type of Streptococcus suis were studied by cefammoximes in vivo. The results of the study were as follows:1. The results showed that the MIC and in vitro bactericidal curves of cefixime to different initial bacteria (normal bacterial amount:106 log10CFU and high colony:108 log10CFU) were not significant, and the results showed that the difference of the MIC between the normal and high strains was not significant. The in vitro sterilization curve shows that the high-bacteria amount inhibits the sterilization effect of the cefixime, and the effect of the fungus quantity is present. The result of the determination of the anti-mutation concentration showed that the anti-mutation concentration was 0.12 & mu; g/ m L, the mutation window of drug resistance was 0.03-0.12 & mu; g/ m L, the selection index was 4, and the ratio was low. In vitro experiments show that the effect of the change of the amount of the bacteria on the MIC is not obvious, but the in vitro sterilization effect is influenced; and the selection of the mutant window indicates that the resistant mutant is not easy to be selected. The pharmacokinetics of cefixime in mice infected with infected mice were studied to establish a local infection model of the thigh of an immune-deficient mouse, which was treated at a dose of 4-fold, and the concentration of the drug was measured by the LC-MS/ MS method before and after administration and after the puncture interval of the vein of the mouse eye frame. The plasma concentration-time data was processed by the pharmacokinetic software of Win Nonlin5.2.1. The results showed that the pharmacokinetics of cefixime in the infected mice were in accordance with the first-order absorption of one-compartment model. The peak concentration of cemedoxime is 1.96-606.71. mu. g/ m L, the area under the curve is 1.58-551.4. m u.g. h/ m L, the peak time is 0.25-0.28 h, the elimination half-life is 0.32-0.38h, and the foundation is laid for the follow-up pharmacodynamics. In order to reduce the interference of the complex bacteria, the antibacterial effect of the cefixime on the type II streptococcus suis is optimized and the selective culture medium is prepared in this paper: polymyxin B (60:30,. mu.g/ m L:. mu.g/ m L) is used in the in vivo efficacy study. An in vivo anti-bacterial effect (PAE) experiment was carried out using an immune-deficient mouse thigh local infection model. The results showed that PAE from 2.5 mg/ kg and 40 mg/ kg was 2.45 h and 8.55 h, respectively, with a concentration-dependent manner. The results show that cefammoximes can be used to produce long PAE in type II Streptococcus suis, and can be used for the treatment of Streptococcus suis with short PAE. This is the first time to reveal the long PAE of the class II Streptococcus suis in vivo, which is the first to reveal the long PAE.4 of the class II Streptococcus suis. The model of local infection in the thigh of mice was studied by the model of drug action/ drug effect of the local infection model of the thigh, and the infection model was made with the normal amount of the local infection and the amount of high bacteria in the thigh of the mouse. The most relevant PK/ PD index, the index range, and the presence of a bacterial volume effect were evaluated by the dose distribution method. The results showed that the antibacterial activity and PK/ PD index (%) of cefammoximes in the two strains of Streptococcus suis were measured. The strongest correlation between the TMICs; the exponential range: the% required to achieve the static effect of two bacterial amounts? When the TMIC does not differ to achieve a 1-log sterilization effect, the% of the amount of high-bacteria infection is infected? TMIC is 1.2 times higher than that of normal bacteria; according to the proportion of the dosage, the proportion of the dose to the static effect and the 1-log sterilization effect is 1.59 and 4.47 times the amount of the normal bacteria, and the result shows that the effect of the ceftrioxime is affected by the amount of the infected bacteria. and then combined with the medicinal kinetic data of the cefixime in the pig body, and through the Monte Carlo analysis, the recommended dosage of the therapeutic clinical pig-infected type II streptococcus suis is as follows: when the MIC is 0.12 & mu; g/ m L, the recommended dose is 2 mg/ kg/24 h i.m., when the MIC is 0.24. mu. g/ m L, The recommended administration dose is 2 mg/ kg/12 h i.m. The results indicated above indicate that Ceftrioxime is used in the clinical treatment of type II swine streptococcal infection, and the dosing protocol should be adjusted according to the MIC value and the degree of infection. The drug action/ drug effect synchronization model of the mouse sepsis model studies the infection of the pigs and the human being caused by the type II streptococcus suis mainly to be septicemia, and therefore, the mice septicemia model is established to further evaluate the drug effect. The results showed that the effective amount of the drug was 2.87 mg/ kg and the% of TMIC was 22.5% ~ 42.5%. It was shown that the Ceftdoxime can also achieve a good therapeutic effect in the model of mice infected with septicemia. In this study, in vitro and in vivo PK/ PD model of mice, the antibacterial activity of cefammoximes on type II swine streptococci was studied, and the most relevant PK/ PD parameters were determined, and the PK/ PD parameter values required for different antibacterial effects were calculated. In order to provide scientific basis for clinical experience treatment, the best treatment plan of swine-infected type II streptococcus suis is recommended by Monte-Carlo analysis in combination with the drug-moving data of cefixime in pig.
【学位授予单位】：华南农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S859.7
[Abstract]:Streptococcus suis is a common animal, and poses a serious threat to the pig industry and public health. The second type of Streptococcus suis is the most popular and most highly pathogenic serotype. The present invention relates to a method for treating type II streptococcus suis infection, and is an ideal choice for treating type II streptococcus suis infection. At present, the study on the drug effect of type II streptococcus suis has not been reported, so it is not possible to provide scientific basis for the treatment of type II streptococcus suis infection. In this paper, the effect of cefixime on the in vitro and in vivo drug effect of Streptococcus suis is studied in this paper. The results of the study on the PK/ PD parameters, the post-antibiotic effect and the amount of the bacteria in the second type of Streptococcus suis were studied by cefammoximes in vivo. The results of the study were as follows:1. The results showed that the MIC and in vitro bactericidal curves of cefixime to different initial bacteria (normal bacterial amount:106 log10CFU and high colony:108 log10CFU) were not significant, and the results showed that the difference of the MIC between the normal and high strains was not significant. The in vitro sterilization curve shows that the high-bacteria amount inhibits the sterilization effect of the cefixime, and the effect of the fungus quantity is present. The result of the determination of the anti-mutation concentration showed that the anti-mutation concentration was 0.12 & mu; g/ m L, the mutation window of drug resistance was 0.03-0.12 & mu; g/ m L, the selection index was 4, and the ratio was low. In vitro experiments show that the effect of the change of the amount of the bacteria on the MIC is not obvious, but the in vitro sterilization effect is influenced; and the selection of the mutant window indicates that the resistant mutant is not easy to be selected. The pharmacokinetics of cefixime in mice infected with infected mice were studied to establish a local infection model of the thigh of an immune-deficient mouse, which was treated at a dose of 4-fold, and the concentration of the drug was measured by the LC-MS/ MS method before and after administration and after the puncture interval of the vein of the mouse eye frame. The plasma concentration-time data was processed by the pharmacokinetic software of Win Nonlin5.2.1. The results showed that the pharmacokinetics of cefixime in the infected mice were in accordance with the first-order absorption of one-compartment model. The peak concentration of cemedoxime is 1.96-606.71. mu. g/ m L, the area under the curve is 1.58-551.4. m u.g. h/ m L, the peak time is 0.25-0.28 h, the elimination half-life is 0.32-0.38h, and the foundation is laid for the follow-up pharmacodynamics. In order to reduce the interference of the complex bacteria, the antibacterial effect of the cefixime on the type II streptococcus suis is optimized and the selective culture medium is prepared in this paper: polymyxin B (60:30,. mu.g/ m L:. mu.g/ m L) is used in the in vivo efficacy study. An in vivo anti-bacterial effect (PAE) experiment was carried out using an immune-deficient mouse thigh local infection model. The results showed that PAE from 2.5 mg/ kg and 40 mg/ kg was 2.45 h and 8.55 h, respectively, with a concentration-dependent manner. The results show that cefammoximes can be used to produce long PAE in type II Streptococcus suis, and can be used for the treatment of Streptococcus suis with short PAE. This is the first time to reveal the long PAE of the class II Streptococcus suis in vivo, which is the first to reveal the long PAE.4 of the class II Streptococcus suis. The model of local infection in the thigh of mice was studied by the model of drug action/ drug effect of the local infection model of the thigh, and the infection model was made with the normal amount of the local infection and the amount of high bacteria in the thigh of the mouse. The most relevant PK/ PD index, the index range, and the presence of a bacterial volume effect were evaluated by the dose distribution method. The results showed that the antibacterial activity and PK/ PD index (%) of cefammoximes in the two strains of Streptococcus suis were measured. The strongest correlation between the TMICs; the exponential range: the% required to achieve the static effect of two bacterial amounts? When the TMIC does not differ to achieve a 1-log sterilization effect, the% of the amount of high-bacteria infection is infected? TMIC is 1.2 times higher than that of normal bacteria; according to the proportion of the dosage, the proportion of the dose to the static effect and the 1-log sterilization effect is 1.59 and 4.47 times the amount of the normal bacteria, and the result shows that the effect of the ceftrioxime is affected by the amount of the infected bacteria. and then combined with the medicinal kinetic data of the cefixime in the pig body, and through the Monte Carlo analysis, the recommended dosage of the therapeutic clinical pig-infected type II streptococcus suis is as follows: when the MIC is 0.12 & mu; g/ m L, the recommended dose is 2 mg/ kg/24 h i.m., when the MIC is 0.24. mu. g/ m L, The recommended administration dose is 2 mg/ kg/12 h i.m. The results indicated above indicate that Ceftrioxime is used in the clinical treatment of type II swine streptococcal infection, and the dosing protocol should be adjusted according to the MIC value and the degree of infection. The drug action/ drug effect synchronization model of the mouse sepsis model studies the infection of the pigs and the human being caused by the type II streptococcus suis mainly to be septicemia, and therefore, the mice septicemia model is established to further evaluate the drug effect. The results showed that the effective amount of the drug was 2.87 mg/ kg and the% of TMIC was 22.5% ~ 42.5%. It was shown that the Ceftdoxime can also achieve a good therapeutic effect in the model of mice infected with septicemia. In this study, in vitro and in vivo PK/ PD model of mice, the antibacterial activity of cefammoximes on type II swine streptococci was studied, and the most relevant PK/ PD parameters were determined, and the PK/ PD parameter values required for different antibacterial effects were calculated. In order to provide scientific basis for clinical experience treatment, the best treatment plan of swine-infected type II streptococcus suis is recommended by Monte-Carlo analysis in combination with the drug-moving data of cefixime in pig.
【学位授予单位】：华南农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S859.7
【参考文献】
相关期刊论文 前8条
1 廖雪玲;张炳旭;丁焕中;;基于防突变浓度的新型药动学-药效学-突变选择窗同步模型在临床用药上的指导作用[J];中国畜牧兽医;2013年01期
2 李敬来;崔孟s，
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