头孢泊肟酯牛至油纳米乳的研制
发布时间：2019-07-05 14:13
【摘要】：目的:为研究头孢泊肟酯与牛至油在纳米水平的联合药效,首次制备头孢泊肟酯牛至油纳米乳(cefpodoxime proxetil and origanum oil nanoemulsion,CFP-OR-NE),并进行药物稳定性、有效性、安全性评价,为治疗致病性大肠杆菌病和赛鸽仙台沙门氏菌病提供一种安全有效的新型口服药物。方法:(1)CFP-OR-NE处方筛选:采用相转变法筛选初步处方。通过加水滴定至相变点并结合伪三元相图的绘制,筛选CFP-OR-NE的初步处方;采用多指标正交试验法筛选CFP-OR-NE最优处方。以L9(34)正交表设计试验,以抑菌圈直径和纳米乳稳定常数Ke为考察指标,选择头孢泊肟酯和牛至油质量分数比(A)、纳米乳水相pH值(B)、乳化温度(C)作为考察因素,筛选头孢泊肟酯牛至油纳米乳最优处方。(2)CFP-OR-NE的质量评价:以多波长紫外分光光度法建立CFP-OR-NE有效成分含量测定方法,进行回收率试验、进样重复性试验及日内日间精密度试验;稳定性试验中考察了CFP-OR-NE结构类型、形态观察、粒径分布及Zeta电位等指标,长期试验中分别建立两者含量-时间回归方程,计算药物有效期。(3)CFP-OR-NE的体外抑菌试验:通过微量肉汤稀释法测定CFP-OR-NE对多种革兰氏阴性菌(G+)和革兰氏阳性菌(G-)的最低抑菌浓度(minimum inhibitory concentration,MIC)和最低杀菌浓度(minimal bactericidal concentration,MBC)。(4)CFP-OR-NE的安全性评价:通过最大耐受计量法测定小鼠经口急性毒性最大耐受剂量并对其进行组织病理学检查,确定CFP-OR-NE是否具有肝毒性、肾毒性、肠毒性。结果:(1)CFP-OR-NE处方筛选:CFP-OR-NE初步处方各组分质量分数为:CFP1.5%、OR1.5%、EL-40 22.51%、1,2-丙二醇1.13%、蒸馏水73.36%;CFP-OR-NE针对致病性大肠杆菌、沙门氏菌最优处方为CFP 0.33%、OR 2.67%、1,2-丙二醇1.13%、EL-40 22.51%、蒸馏水73.36%,纳米乳pH值为6.17。(2)质量评价:建立了CFP-ORNE药物含量测定方法,CFP测定的检测波长和参比波长分别为263nm和289.8nm,OR测定的组合波长为261.6 nm、273.2 nm、284.4 nm。在1~100μg/mL范围内,CFP和OR线性关系均良好(r2=0.999 22;r2=0.999 70);回收率试验:9个试验号的回收率均符合70%~110%的要求,CFP的RSD范围在0.4~8.95%之间,OR的RSD范围在0.26~5.57%之间;进样重复性试验:各试验号CFP及OR测定的RSD均小于0.6%,进样重复性良好;日内日间精密度试验:CFP-OR-NE中CFP和OR测定的日内、日间精密度RSD均小于5%,日内日间精密度良好;CFP-OR-NE为水包油型淡黄色透明液体,形态观察CFPOR-NE乳滴呈圆球形,平均粒径为18.94 nm,多分散系数0.258,粒径大小均一且分布范围较窄。在环境温度25℃条件下,稀释5倍后的平均pH值为6.39,zeta电位是(-9.96±6.35)mV,加速、离心、光照、温度梯度实验、长期试验结果均无分层、絮凝、转型、破乳、酸败等现象,测定CFP-OR-NE有效期为21个月。(3)体外抑菌试验:与OR原料药、头孢克肟(cefixime,CFM)原料药相比,CFP-OR-NE的MIC及MBC差异显著(P0.05)。(4)安全性评价:最大耐受计量法测定小鼠经口急性毒性最大耐受剂量大于1 800 mg/kg,组织病理学检查CFP-OR-NE无肾脏毒性和肠毒性,试验组肝脏,轻微出血,少许细胞变性坏死,分界不清,但未引起小鼠死亡。结论:研制出CFP-OR-NE并建立了药物含量测定方法,质量评价符合规定要求;体外抑菌试验表明CFP-OR-NE对临床常见G+和G-的抑菌和杀菌效果优于同为三代头孢菌素的CFM;MTD试验表明CFP-OR-NE为低毒药物,组织病理学检查无肾、肠毒性。
[Abstract]:Objective: To study the effect of cefpodoxime proxetil (Cefpodoxime proxetil) and bovine-to-oil at nanometer level, and to prepare cefpodoxime and oil nanoemulsion (CFP-OR-NE) for the first time, and to evaluate the stability, effectiveness and safety of cefpodoxime proxetil (CFP-OR-NE). In ord to provide a safe and effective novel oral medicine for treating that pathogenic E. coli and the salmonellosis. Methods: (1) CFP-OR-NE prescription screening: a phase transition method was used to screen the initial prescription. The initial formulation of CFP-OR-NE was selected by titration of water to the phase change point and the pseudo-ternary phase diagram, and the optimal formulation of the CFP-OR-NE was selected by the multi-index orthogonal test method. The experiment was designed with L9 (34) orthogonal table, and the diameter of the bacteriostatic ring and the stability constant of the nano-emulsion Ke were selected as the study index, and the ratio (A), the pH value (B) of the nano-emulsion water phase and the emulsification temperature (C) were selected as the factors of the investigation. The optimal formulation of the cefpodoxime proxetil to the oil nanoemulsion is screened. (2) The quality evaluation of CFP-OR-NE: The determination method of the effective component of CFP-OR-NE was established by multi-wavelength ultraviolet spectrophotometry, the recovery test, the injection repeatability test and the day-day precision test were carried out, and the structure type and morphology of the CFP-OR-NE were investigated in the stability test. In the long-term test, the content-time regression equation was established, and the validity of the drug was calculated. (3) In vitro antibacterial test of CFP-OR-NE: the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CFP-OR-NE on a variety of Gram-negative bacteria (G +) and gram-positive bacteria (G-) were determined by a microbroth dilution method. (4) Safety evaluation of CFP-OR-NE: The maximum tolerated dose of the oral acute toxicity of the mouse was determined by the maximum tolerance measurement method and the histopathological examination was performed to determine whether the CFP-OR-NE had hepatotoxicity, renal toxicity, and intestinal toxicity. Results: (1) CFP-OR-NE prescription was selected: CFP1.5%, OR1.5%, EL-40 22.51%, 1,2-propanediol 1.13%, distilled water 73.36%, CFP-OR-NE for pathogenic E. coli, the optimal prescription of Salmonella was CFP 0.33%, OR 2.67%, 1,2-propanediol 1.13%, EL-40 22.51%, The distilled water was 73.36%, and the pH value of the nano-emulsion was 6.17. (2) Quality evaluation: The method for determination of the content of CFP-ORNE was established. The detection wavelength and reference wavelength of CFP were 263 nm and 289.8 nm, respectively, and the combined wavelength was 261.6 nm, 273.2 nm and 284.4 nm respectively. In the range of 1 to 100 & mu; g/ mL, the linear relationship between the CFP and the OR was good (r2 = 0.999 22; r2 = 0.999 70); the recovery test: the recovery rate of the 9 test numbers met the requirements of 70% to 110%, the RSD of the CFP was between 0.4 and 8.95%, the RSD of the OR was between 0.26 and 5.57%, and the injection repeatability test: The RSD of each test number CFP and OR was less than 0.6% and the injection repeatability was good; intra-day precision test: the day-day precision RSD was less than 5% within the day of the CFP and OR measurement in the CFP-OR-NE, and the intra-day precision was good; and the CFP-OR-NE is an oil-in-water type light yellow transparent liquid, The form of CFPOR-NE emulsion is spherical, the average particle size is 18.94 nm, the polydispersity coefficient is 0.258, the particle size is uniform and the distribution range is narrow. The mean pH value after 5-fold dilution at ambient temperature was 6.39, the zeta potential was (-9.96-6.35) mV, acceleration, centrifugation, illumination, temperature gradient experiment, and the results of long-term test were no delamination, flocculation, transformation, demulsification, rancidity, etc., and the validity period of CFP-OR-NE was 21 months. (3) In vitro antibacterial test: Compared with the drug substance of OR drug substance and cefixime (CFM), the MICs and MBC of CFP-OR-NE were significant (P0.05). (4) Safety evaluation: The maximum tolerated dose of the oral acute toxicity in mice was more than 1 800 mg/ kg, and the histopathological examination of CFP-OR-NE had no renal toxicity and intestinal toxicity. Conclusion: CFP-OR-NE has been developed and the drug content determination method is established, and the quality evaluation is in accordance with the specified requirements; in vitro antibacterial test shows that the antibacterial and bactericidal effect of CFP-OR-NE on the clinical common G + and G-is better than that of the three-generation cephalosporin; and the MTD test shows that the CFP-OR-NE is a low-toxic medicine, Histopathological examination showed no renal or intestinal toxicity.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S859.79
本文编号：2510581
[Abstract]:Objective: To study the effect of cefpodoxime proxetil (Cefpodoxime proxetil) and bovine-to-oil at nanometer level, and to prepare cefpodoxime and oil nanoemulsion (CFP-OR-NE) for the first time, and to evaluate the stability, effectiveness and safety of cefpodoxime proxetil (CFP-OR-NE). In ord to provide a safe and effective novel oral medicine for treating that pathogenic E. coli and the salmonellosis. Methods: (1) CFP-OR-NE prescription screening: a phase transition method was used to screen the initial prescription. The initial formulation of CFP-OR-NE was selected by titration of water to the phase change point and the pseudo-ternary phase diagram, and the optimal formulation of the CFP-OR-NE was selected by the multi-index orthogonal test method. The experiment was designed with L9 (34) orthogonal table, and the diameter of the bacteriostatic ring and the stability constant of the nano-emulsion Ke were selected as the study index, and the ratio (A), the pH value (B) of the nano-emulsion water phase and the emulsification temperature (C) were selected as the factors of the investigation. The optimal formulation of the cefpodoxime proxetil to the oil nanoemulsion is screened. (2) The quality evaluation of CFP-OR-NE: The determination method of the effective component of CFP-OR-NE was established by multi-wavelength ultraviolet spectrophotometry, the recovery test, the injection repeatability test and the day-day precision test were carried out, and the structure type and morphology of the CFP-OR-NE were investigated in the stability test. In the long-term test, the content-time regression equation was established, and the validity of the drug was calculated. (3) In vitro antibacterial test of CFP-OR-NE: the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CFP-OR-NE on a variety of Gram-negative bacteria (G +) and gram-positive bacteria (G-) were determined by a microbroth dilution method. (4) Safety evaluation of CFP-OR-NE: The maximum tolerated dose of the oral acute toxicity of the mouse was determined by the maximum tolerance measurement method and the histopathological examination was performed to determine whether the CFP-OR-NE had hepatotoxicity, renal toxicity, and intestinal toxicity. Results: (1) CFP-OR-NE prescription was selected: CFP1.5%, OR1.5%, EL-40 22.51%, 1,2-propanediol 1.13%, distilled water 73.36%, CFP-OR-NE for pathogenic E. coli, the optimal prescription of Salmonella was CFP 0.33%, OR 2.67%, 1,2-propanediol 1.13%, EL-40 22.51%, The distilled water was 73.36%, and the pH value of the nano-emulsion was 6.17. (2) Quality evaluation: The method for determination of the content of CFP-ORNE was established. The detection wavelength and reference wavelength of CFP were 263 nm and 289.8 nm, respectively, and the combined wavelength was 261.6 nm, 273.2 nm and 284.4 nm respectively. In the range of 1 to 100 & mu; g/ mL, the linear relationship between the CFP and the OR was good (r2 = 0.999 22; r2 = 0.999 70); the recovery test: the recovery rate of the 9 test numbers met the requirements of 70% to 110%, the RSD of the CFP was between 0.4 and 8.95%, the RSD of the OR was between 0.26 and 5.57%, and the injection repeatability test: The RSD of each test number CFP and OR was less than 0.6% and the injection repeatability was good; intra-day precision test: the day-day precision RSD was less than 5% within the day of the CFP and OR measurement in the CFP-OR-NE, and the intra-day precision was good; and the CFP-OR-NE is an oil-in-water type light yellow transparent liquid, The form of CFPOR-NE emulsion is spherical, the average particle size is 18.94 nm, the polydispersity coefficient is 0.258, the particle size is uniform and the distribution range is narrow. The mean pH value after 5-fold dilution at ambient temperature was 6.39, the zeta potential was (-9.96-6.35) mV, acceleration, centrifugation, illumination, temperature gradient experiment, and the results of long-term test were no delamination, flocculation, transformation, demulsification, rancidity, etc., and the validity period of CFP-OR-NE was 21 months. (3) In vitro antibacterial test: Compared with the drug substance of OR drug substance and cefixime (CFM), the MICs and MBC of CFP-OR-NE were significant (P0.05). (4) Safety evaluation: The maximum tolerated dose of the oral acute toxicity in mice was more than 1 800 mg/ kg, and the histopathological examination of CFP-OR-NE had no renal toxicity and intestinal toxicity. Conclusion: CFP-OR-NE has been developed and the drug content determination method is established, and the quality evaluation is in accordance with the specified requirements; in vitro antibacterial test shows that the antibacterial and bactericidal effect of CFP-OR-NE on the clinical common G + and G-is better than that of the three-generation cephalosporin; and the MTD test shows that the CFP-OR-NE is a low-toxic medicine, Histopathological examination showed no renal or intestinal toxicity.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S859.79
【参考文献】
相关期刊论文 前1条
1 张涛;顾起有;杨后贵;郭杨庆;;头孢泊肟酯与头孢克肟治疗急性细菌性感染随机对照研究[J];海峡药学;2006年02期
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