【摘要】：目的研究中藥有效成分補(bǔ)骨脂素和異補(bǔ)骨脂素的細(xì)胞色素P450(CYP)酶促動(dòng)力學(xué)特征,比較其結(jié)構(gòu)和種屬差異,為體內(nèi)藥代動(dòng)力學(xué)特征的預(yù)測(cè)提供科學(xué)依據(jù)。方法建立補(bǔ)骨脂素和異補(bǔ)骨脂素的液相色譜串聯(lián)質(zhì)譜(LC-MS/MS)檢測(cè)方法,在優(yōu)化人和大鼠肝微粒體孵育體系和評(píng)價(jià)體外代謝穩(wěn)定性的基礎(chǔ)上,進(jìn)行了代謝穩(wěn)定性和酶促動(dòng)力學(xué)研究,應(yīng)用非線性回歸法計(jì)算最大反應(yīng)速率(V_(max))和米氏常數(shù)(K_m)。結(jié)果應(yīng)用建立的LC-MS/MS檢測(cè)方法,補(bǔ)骨脂素、異補(bǔ)骨脂素在肝微粒體孵育液中定量范圍為0.1~50.0μmol·L~(-1),線性關(guān)系良好,精密度和準(zhǔn)確度等滿足檢測(cè)要求。體外代謝穩(wěn)定性研究顯示,當(dāng)?shù)孜餄舛葹?μmol·L~(-1),蛋白濃度為0.5 g·L~(-1),孵育40 min內(nèi),補(bǔ)骨脂素和異補(bǔ)骨脂素在大鼠和人肝微粒體呈線性消除,體外半衰期分別為74.5,95.0,74.5和173.3 min。補(bǔ)骨脂素在大鼠和人肝微粒體中的V_(max)分別為:(1.140±0.080)μmol·min-1·g-1蛋白,(0.620±0.060)μmol·min-1·g-1蛋白;K_m分別為(12.9±0.3)μmol·L~(-1)和(7.4±1.3)μmol·L~(-1)。異補(bǔ)骨脂素在大鼠和人肝微粒體中的V_(max)分別為(0.251±0.012)μmol·min-1·g-1蛋白和(0.103±0.014)μmol·min-1·g-1蛋白;K_m分別為:(3.0±0.4)μmol·L~(-1)和(3.4±0.7)μmol·L~(-1)。結(jié)論補(bǔ)骨脂素和異補(bǔ)骨脂素的CYP酶促動(dòng)力學(xué)特征及代謝穩(wěn)定性具有一定的種屬和結(jié)構(gòu)差異,在大鼠兩者基于CYP酶的代謝清除過(guò)程可能相似。而在人體,異補(bǔ)骨脂素的CYP代謝清除可能慢于補(bǔ)骨脂素,導(dǎo)致藥代動(dòng)力學(xué)特征的差異。
[Abstract]:Objective to study the enzyme kinetic characteristics of pigment P450 (CYP) of psoralen and isopsoralen, and to compare their structure and species differences, so as to provide scientific basis for the prediction of pharmacokinetics characteristics in vivo. Methods A liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the determination of psoralen and isopsoralen was established. On the basis of optimizing the incubation system of human and rat liver microsomes and evaluating the metabolic stability in vitro, the metabolic stability and enzyme kinetics were studied. The maximum reaction rates (V _ (max) and K 鈮，

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