【作者】 李振；

【导师】 宋晓平；

【作者基本信息】 西北农林科技大学 ， 临床兽医学， 2011， 硕士

【摘要】 羟甲香豆素（hymecromone）,化学名为7-羟基-4-甲基-2H-1-苯并吡喃-2-酮,被广泛应用于工业、农业、医药等领域。在医药领域,羟甲香豆素主要作为利胆药用于治疗胆囊炎、胆石症、胆道感染等。本实验室从菊科蒿属植物猪毛蒿（Artemisia scoparia Walds et kit）中提取得到具有高效杀螨活性的单体,结构鉴定为羟甲香豆素。羟甲香豆素属脂水难溶性化合物,国外对羟甲香豆素在人和大鼠体内的药代动力学研究均使用其钠盐,本试验首先进行羟甲香豆素钠盐的制备,建立家兔血浆内羟甲香豆素含量的反相高效液相测定法,进而研究羟甲香豆素在家兔体内的药代动力学规律。结果如下:1.羟甲香豆素钠盐的制备:以10 mmol羟甲香豆素与9.5 mmol氢氧化钠为原料,92%的丙酮水溶液进行重结晶得到淡黄色针状晶体,产率75%。采用TLC,UV,IR,¹H NMR,¹³C NMR和MS进行结构表征,确定所得化合物为羟甲香豆素钠盐;建立测定样品中羟甲香豆素钠盐含量的紫外分光光度法,标准曲线方程为y=0.0869x-0.3011（n=7）,R²=0.9976,在5³0μg/mL浓度范围内,线性关系良好。制备的样品中羟甲香豆素钠盐含量为98%。2.家兔血浆中羟甲香豆素含量测定的RP-HPLC方法的建立:采用C18柱（4.6×150mm, 5μm）,甲醇-水（55:45）为流动相,7-羟基香豆素为内标,乙腈沉淀蛋白,检测波长324 nm,柱温30℃,测定家兔血浆中的羟甲香豆素。在此色谱条件下,羟甲香豆素浓度在20³2000 ng/mL范围内与峰面积比呈良好的线性关系,标准曲线方程为y=0.0005x-0.0036（n=10）,R²=0.9998。日内和日间精密度（n=5）RSD<4%,平均回收率为100.02%,萃取回收率为95.78%。该法灵敏、简便、准确、快捷,特异性强,适用于羟甲香豆素在家兔体内的药代动力学研究。3.羟甲香豆素在家兔体内的药代动力学研究:应用RP-HPLC方法研究静脉注射和经皮给药后,羟甲香豆素在家兔体内的药代动力学过程。结果表明,羟甲香豆素在家兔体内的药代动力学符合二室模型。静脉注射后,平均药动学方程为:C=14997.19e^-3.957t+ 161.72e^-0.05t;药时曲线下面积（AUC）=3967.28±1554.60 ng/mL·h;分布半衰期(T_1/2α)=0.18±0.04 h;消除半衰期(T_1/2β)=15.99±5.25 h;表观分布容积（Vd）=5.19±2.10 L/（h·kg）。经皮给药后,平均药动学方程为C=210.13e^-0.31+108.02e^-0.0032t-318.15e^-4.06t;AUC= 9295.16±3411.64 ng/mL·h;吸收半衰期（T1/2kα）=0.18±0.05 h;T_1/2α=2.27±0.24 h; T_1/2β=54.92±14.93 h;峰浓度(C_max)=318.15±4367 ng/mL; Vd=493.21±65.29 L/（h·kg）; CL=2030.62±691.41 ng/（kg·h）。更多还原

【Abstract】 Hymecromone, the chemical name is 7- hydroxy -4- methyl-2H-1-Benzopyran-2- ketone, is widely used in industry, agriculture, medicine and other fields. In the ield of medicine, hymecromone are primarily used as a choleretic drug for the treatment of cholecystitis, cholelithiasis, biliary tract infections, gallbladder surgery syndrome, etc. A monomer which has a highly efficient of acaricidal activity has benn extracted from Artemisia scoparia Walds et kit, and it was identified as hymecromone. Hymecromone is insoluble in water and fat, its sodium salt was used in pharmacokinetics studies on hymecromone in humans and rats abroad. In this study, the hymecromone sodium salt was synthesized firstly, and a method of determining the concentration of hymecromone of blood plasma by reversed-phase high performance liquid chromatography in rabbits was established. Then, the pharmacokinetics of hymecromone in rabbits were studied. The results were as follows:1. Synthesis of hymecromone sodium salt: Hymecromone sodium salt was prepared by reaction of hymecromone （10 mmol） with sodium hydroxide（9.5 mmol）, recrystallization in the solution of 92% acetone in water provided pale yellow needle crystal with yield of 75%, The structure of the compound was characterized by TLC, UV, IR, 1H NMR, 13C NMR and MS, and was confirmed to be hymecromone sodium salt. The content of the product samples detected by the UV method was built, which the equation of standard curves was y=0.0869x-0.3011 （n=7）, （R²=0.9976） and the linear relations were good in the concentration range of 5³0μg/mL, and the mean content of hymecromone sodium salt synthesized by this new process was 98%.2. A Method for determination content of hymecromone in rabbits blood plasma was established: the chromatographic conditions, which was used for determinating the content of hymecromone in rabbits blood plasma, were decided to be C18 column （4.6×150 mm, 5μm）, Methanol and water （55:45） as mobile phase, 7-hydroxy coumarin as the internal standard and acetonitrile precipitation method for removing albumin with detective UV wavelength at 324 nm and column temperature at 30℃. Under these eonditions, the ratio of peak areas for hymecromone to internal standard 7-hydroxy coumarin was linear with the content of hymecromone in the range of 20³2000 ng/mL. The RSD of with-in day precision test and between-day precision test （n=5）were both less 4%. The average recovery and extraction recovery was 100.02% and 95.78%. This method is sensitive, simple, accurate, fast, specific for pharmacokinetics study on hymecromone in rabbits.3. Pharmacokinetics study on hymecromone in rabbits: The pharmaeokineties study of hymecromone in rabbits was made after intravenous injection and transdermal drug delivery using RP-HPLC method. The results showed that hymecromone abided by two-compartment model in rabbits. After intravenous injection, its average pharmacokinetic equations was C=14997.19e^-3.957t+161.72e^-0.05t; the area under the plasma concentration-time curve （AUC） was 3967.28±1554.60 ng/mL·h, the distribution half-life (T_1/2α) was 0.18±0.04h, the elimination half-life (T_1/2β) was 15.99±5.25 h, the apparent volume of distribution （Vd） was 5.19±2.10 L/（h·kg）. After transdermal drug delivery, its average pharmacokinetic equations was C=210.13e^-0.31+108.02e^-0.0032t-318.15e^-4.06t; AUC was 9295.16±3411.64 ng/mL·h, the absorption half-life （T1/2kα） was 0.18±0.05 h, T_1/2α was 2.27±0.24 h, T_1/2β was 54.92±14.93 h, the peak concentration (C_max) was 318.15±4367 ng/mL, Vd was 493.21±65.29 L/（h·kg）, the CL was 2030.62±691.41 ng/（kg·h）.更多还原