以液相層析分析法測定新抗癌藥物 1-[3-(Furo[3,2-C]quinolin-4-ylamino)phenyl] ethanoe-O-methyl- oxime 於大白鼠之藥物動力學   全文下載

Pharmacokinetics of A New Antitumor Agent, 1-[3-(Furo[3,2-C]quinolin-4-ylamino)phenyl]-ethanoe -O-methyl-oxime, in Rat Using A High Performance Liquid Chromatography Method

1-[3-（Furo[3,2-C]quinolin-4-ylamino）phenyl]-ethanoe-O-methyl-oxime（CCK3）是一種新合成的抗癌藥，經美國癌症研究中心（NCI）進行抗癌活性篩選後發現其對特定的的細胞株如腎癌細胞的UO-31 和黑色素癌細胞的UACC-257 及UACC-62有特殊的活性；而且由其結構活性相關性推測其半衰期應較amsacrine（約30 分鐘）為長。所以本研究之目的在於開發出一種高感度的HPLC 方法，以測定CCK3 於動物體中之藥物動力學，以評估CCK3日後發展成為臨床用藥之可行性。在分析方法開發中，將CCK3加入空白大白鼠血漿中以dichloromethane抽提後，以逆相層析管柱分析。沖溶條件如次：移動相為乙A30%， tetrahydrofuran 5% 和pH 3.0 磷酸鹽緩衝溶液65%；流速每分鐘1.0 mL 進行；檢品以電化學檢出器於電壓1.0 伏特和電流10 nA 檢定；內部標準品為2-naphthol 。在分析方法確認方面，在特定濃度範圍內10- 3,000 ng/mL ，其分析之相對標準差和相對誤差均小於10%；而且在此設定濃度範圍下其濃度與得到之相關面積比均有良好的線性關係，顯示此分析方法可應用於定量分析。CCK的最低定量濃度（LOQ）和最低偵測濃度（LOD）分別為10 ng/mL 和5 ng/mL 。在體內實驗方面，以靜脈注射方式，給予大白鼠3.97 和7.94 mg/kg CCK3 二個劑量，結果顯示其分佈體積、半衰期和清除率並不受劑量多寡影響，而AUC則隨劑量加倍而成等比例增加，而CCK3在大白鼠體內之半衰期約1.53 hr ，為amsacrine 的3 倍。顯示CCK3可能具有臨床實驗之潛力值得進一步研宄。"

1-[3-(Furo[3,2-C]quinolin-4-ylamino)phenyl]ethanoe-O-methyl-oxime (CCK3) is an antitumor agent, particularly active against the growth of the renal cancer cell, UO-31, and two melanoma cancer cells, UACC-257 and UACC-62, as indicated in the NCI’s full panel of 60 human cancer cell lines cytotoxicity evaluation. From the structure-activity relationships between amsacrine and CCK3, CCK3 was expected to have longer half-life than amsacrine in plasma. Therefore, a reversed-phase high performance liquid chromatography method was developed and validated for the determination of pharmacokinetics of CCK3 in rats. The plasma samples were spiked with the internal standard 2-naphthol and extracted using dichloromethane. A C18 column (55 × 4 mm) was used for the separation of analytes with a mobile phase consisted of 30% acetonitrile, 5% tetrahydrofuran and 65% pH 3.0 of McIlvaine buffer at a flow rate of 1.0 mL/min. CCK3 was detected and utilized by electrochemical detector at 1.0 voltage and 10 nA. Intra- and inter-day precision and accuracy were acceptable down to the limit of quantitation of 10 ng/mL. The lower limit of detection was 5 ng/mL. As for the in vivo study, the pharmacokinetic parameters of CCK3 in rats after intravenous administration of 3.97 and 7.94 mg/kg were determined. The apparent volume of distribution, half-life and clearance showed no significant difference between these two dosages. The area under the plasma concentration time curve increased proportionally with increase in dose. The half-life of CCK3 was prolonged 3 folds, compared to amsacrine. Therefore, CCK3 might have the potential to be tested clinically.