毛萼乙素环糊精包合物在大鼠体内的非线性药物代谢动力学研究
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摘要
毛萼乙素(Eriocalyxin B,EriB)是疏花毛萼香茶菜的一个贝壳杉烷二萜类化合物,具有较强的抗肿瘤活性,但毒性较大.本文利用HPLC-UV的定量分析方法,进行不同剂量的EriB环糊精包合物在大鼠体内的药物动力学研究;利用大鼠和人重组肝微粒体,进行CYP450代谢酶研究;利用超滤法进行血浆蛋白结合研究.结果显示:单次静脉注射、腹腔注射给药5 mg/kg、10 mg/kg、15 mg/kg时,EriB在大鼠体内呈非线性代谢.进一步的研究发现EriB能被快速吸收,其血浆蛋白结合率随药物浓度而发生变化,EriB几乎不以原型从尿液和粪便排出体外,可以被CYP450酶剧烈代谢,其中CYP2D6在该过程中起到了最重要的作用,且EriB或其代谢物能显著诱导CYP1A2(14.63%)和CYP3A4(45.54%).总之,EriB的药代动力学是非线性的,可能是其毒性产生的原因之一;血浆蛋白结合率随浓度变化和其对CYP450酶的影响可能是导致非线性代谢的原因.
Eriocalyxin B(EriB) is an ent-kaurane diterpenoid that exhibits promise as a potential broad-spectrum anticancer agent but with toxicity. In this study the pharmacokinetic profile in rats of different EriB doses were assayed with a validated HPLC-UV method. Then its metabolism behaviour was investigated in liver microsomes and the plasma binding rate was tested through ultra-filter method. The results demonstrated that EriB metabolism profile in rats is nonlinear via i.v. or i.p. Further studies found that EriB was absorbed quickly and its plasma protein binding rate changes at different doses. EriB was barely excreted in urine and faeces as prototype. However, more than 90% of EriB was metabolised in rat liver microsomes, and CYP2 D6 played the most important role in this process, according to chemical inhibitor and human recombinant CYP450 enzyme tests. EriB or its metabolites significantly induced CYP1 A2(14.63%) and CYP3 A4(45.54%). In conclusion, the pharmacokinetics of EriB were nonlinear, which may partly contribute to its toxicity; the variety of plasma protein binding rate and its influence on CYP450 might cause the nonlinear pharmacokinetics behaviours.
Eriocalyxin B(EriB) is an ent-kaurane diterpenoid that exhibits promise as a potential broad-spectrum anticancer agent but with toxicity. In this study the pharmacokinetic profile in rats of different EriB doses were assayed with a validated HPLC-UV method. Then its metabolism behaviour was investigated in liver microsomes and the plasma binding rate was tested through ultra-filter method. The results demonstrated that EriB metabolism profile in rats is nonlinear via i.v. or i.p. Further studies found that EriB was absorbed quickly and its plasma protein binding rate changes at different doses. EriB was barely excreted in urine and faeces as prototype. However, more than 90% of EriB was metabolised in rat liver microsomes, and CYP2 D6 played the most important role in this process, according to chemical inhibitor and human recombinant CYP450 enzyme tests. EriB or its metabolites significantly induced CYP1 A2(14.63%) and CYP3 A4(45.54%). In conclusion, the pharmacokinetics of EriB were nonlinear, which may partly contribute to its toxicity; the variety of plasma protein binding rate and its influence on CYP450 might cause the nonlinear pharmacokinetics behaviours.
引文
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[20] 孙汉董,朱兆云,梅双喜,等.毛萼乙素环糊精包合物、含有该包合物的药物组合物、其制备方法及应用:中国,201410007598.7[P].2015-03-23.
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[22] 张荣,刘昌辉,王宁生,等.以睾酮为探针采用高效液相色谱法测定细胞色素CYP450 3A4的酶活性[J].色谱,2008,26(1):80-83.
[23] 王建,侯惠民,夏怡然,等.毛萼乙素纳米混悬剂.I.制备及在小鼠体内的药动学[J].中国医药工业杂志,2011,43(2):100-103.
[24] Li X T,Chu Y L,Wu F J,et al.Determination of plasma protein binding of ponicidin by ultrafiltration[J].Chinese Journal of New Drugs,2013,22(20):2418-2422.
[25] 夏怡然,王健,朱金屏,等.大鼠血浆中毛萼乙素的HPLC测定[J].中国医药工业杂志,2010(1):41-43.
[26] Li L,Yue G G,Lau C B,et al.Eriocalyxin B induces apoptosis and cell cycle arrest in pancreatic adenocarcinoma cells through caspase- and p53-dependent pathways[J].Toxicology & Applied Pharmacology,2012,262(1):80-90.
[27] 王健,侯惠民,夏怡然,等.毛萼乙素纳米混悬剂.II.对小鼠肝癌H22移植瘤的生长抑制作用[J].中国医药工业杂志,2012,43(3):194-197.
[28] 刘建平.生物药剂学与药物动力学[M].北京:人民卫生出版社,2000:263-270.
[29] Ludden T M.Nonlinear Pharmacokinetics[J].Clinical Pharmacokinetics,1992,20(6):429-446.
[30] Iwasaki K,Murayama N,Koizumi R,et al.Comparison of cytochrome P450 3A enzymes in cynomolgus monkeys and humans[J].Drug Metabolism & Pharmacokinetics,2010,25(4):388-391.
[31] Aueviriyavit S,Kobayashi K,Kan C.Species Differences in Mechanism-Based Inactivation of CYP3A in Humans,Rats and Mice[J].Drug Metabolism & Pharmacokinetics,2010,25(1):93-100.
[32] Iwasaki K,Murayama N,Koizumi R,et al.Comparison of cytochrome P450 3A enzymes in cynomolgus monkeys and humans[J].Drug Metabolism & Pharmacokinetics,2010,25(4):388-391.
[33] Usmani K A,Hodgson E,Rose R L.In vitro metabolism of carbofuran by human,mouse,and rat cytochrome P450 and interactions with chlorpyrifos,testosterone,and estradiol[J].Chemico-Biological Interactions,2004,150(3):221-232.
[34] Guo F,An T,Rein K S.The algal hepatoxoxin okadaic acid is a substrate for human cytochromes CYP3A4 and CYP3A5[J].Toxicon,2010,55(2):325-332.
[35] Tang J,Usmani K A,Hodgson E.In vitro metabolism of fipronil by human and rat cytochrome P450 and its interactions with testosterone and diazepam[J].Chemico-Biological Interactions,2004,147(3):319-329.
[2] Wang L,Zhao W L,Yan J S,et al.Eriocalyxin B induces apoptosis of t(8;21) leukemia cells through NF-kappaB and MAPK signaling pathways and triggers degradation of AML1-ETO oncoprotein in a caspase-3-dependent manner[J].Cell Death & Differentiation,2007,14(2):306-317.
[3] Lu Y,Chen B,Song J H,et al.Eriocalyxin B ameliorates experimental autoimmune encephalomyelitis by suppressing Th1 and Th17 cells[J].Pnas,2013,110(6):2258-2263.
[4] Li L,Yue G G,Lau C B,et al.Eriocalyxin B induces apoptosis and cell cycle arrest in pancreatic adenocarcinoma cells through caspase- and p53-dependent pathways[J].Toxicology & Applied Pharmacology,2012,262(1):80-90.
[5] Zhang Y W,Jiang X X,Chen Q S,et al.Eriocalyxin B induces apoptosis in lymphoma cells through multiple cellular signaling pathways[J].Experimental Hematology,2010,38(3):191-201.
[6] Chen R,Alvero A B,Silasi D A,et al.Regulation of IKKbeta by miR-199a affects NF-kappaB activity in ovarian cancer cells[J].Oncogene,2008,27(34):4712-4723.
[7] Li X,Pu J,Jiang S,et al.Henryin,an ent-kaurane diterpenoid,inhibits Wnt signaling through interference with β-Catenin/TCF4 interaction in colorectal cancer cells[J].PLOS ONE,2013,8(7):e68525.
[8] 王健,侯惠民,夏怡然,等.毛萼乙素纳米混悬剂.II.对小鼠肝癌H22移植瘤的生长抑制作用[J].中国医药工业杂志,2012,43(3):194-197.
[9] 孙汉董,赵勤实,牛雪梅,等.一种治疗癌症的药物及其用途:中国,CN1424028[P].2003-06-18.
[10] 任建婷,张洪信,陆顺元,等.中药制剂毛萼乙素对免疫球蛋白类别转换的作用[J].上海交通大学学报(医学版),2013,33(7):949-954.
[11] Leung C H,Grill S P,Lam W,et al.Eriocalyxin B inhibits nuclear factor-kappaB activation by interfering with the binding of both p65 and p50 to the response element in a noncompetitive manner[J].Molecular Pharmacology,2006,70(6):1946-1955.
[12] Gao L W,Zhang J,Yang W H,et al.Glaucocalyxin A induces apoptosis in human leukemia HL-60 cells through mitochondria-mediated death pathway[J].Toxicology in Vitro,2011,25(1):51-63.
[13] Lu Y,Peng Z G,Yuan T T,et al.Multi-sites cleavage of leukemogenic AML1-ETO fusion protein by caspase-3 and its contribution to increased apoptotic sensitivity[J].Leukemia,2008,22(2):378-86.
[14] Rasul A,Liu J,Liu Y,et al.Eriocalyxin B inhibits proliferation and induces apoptosis through downregulation of Bcl-2 and activation of caspase-3 in human bladder cancer cells[J].Bangladesh Journal of Pharmacology,2013,8(2):116-123.
[15] Li L,Yue G G,Pu J X,et al.Eriocalyxin B-Induced Apoptosis in Pancreatic Adenocarcinoma Cells Through Thiol-Containing Antioxidant Systems and Downstream Signalling Pathways[J].Current Molecular Medicine,2014,14(5):673-689.
[16] Yu X K,He L,Cao P,et al.Eriocalyxin B inhibits STAT3 signaling by covalently targeting STAT3 and blocking phosphorylation and activation of STAT3[J].Plos One.2015,10(5):e0128406.
[17] Zhou X,Yue G G,Liu M,et al.Eriocalyxin B,a natural diterpenoid,inhibited VEGF-induced angiogenesis and diminished angiogenesis-dependent breast tumor growth by suppressing VEGFR-2 signaling[J].Oncotarget,2016,7(50):82820-82835.
[18] Liu Y F,Qu G Q,Lu Y M,et al.Silencing of MAP4K4 by short hairpin RNA suppresses proliferation,induces G1 cell cycle arrest and induces apoptosis in gastric cancer cells[J].Molecular medicine reports,2016,13(1):41-48.
[19] Zhou X,Yue G G L,Chan A M L,et al.Eriocalyxin B,a novel autophagy inducer,exerts anti-tumor activity through the suppression of Akt/mTOR/p70S6K signaling pathway in breast cancer[J].Biochemical pharmacology,2017,142:58-70.
[20] 孙汉董,朱兆云,梅双喜,等.毛萼乙素环糊精包合物、含有该包合物的药物组合物、其制备方法及应用:中国,201410007598.7[P].2015-03-23.
[21] 毕惠嫦,和凡,温莹莹,等.丹参酮ⅡA在大鼠体内的药代动力学及相关机制研究[J].中草药,2007,38(6):882-886.
[22] 张荣,刘昌辉,王宁生,等.以睾酮为探针采用高效液相色谱法测定细胞色素CYP450 3A4的酶活性[J].色谱,2008,26(1):80-83.
[23] 王建,侯惠民,夏怡然,等.毛萼乙素纳米混悬剂.I.制备及在小鼠体内的药动学[J].中国医药工业杂志,2011,43(2):100-103.
[24] Li X T,Chu Y L,Wu F J,et al.Determination of plasma protein binding of ponicidin by ultrafiltration[J].Chinese Journal of New Drugs,2013,22(20):2418-2422.
[25] 夏怡然,王健,朱金屏,等.大鼠血浆中毛萼乙素的HPLC测定[J].中国医药工业杂志,2010(1):41-43.
[26] Li L,Yue G G,Lau C B,et al.Eriocalyxin B induces apoptosis and cell cycle arrest in pancreatic adenocarcinoma cells through caspase- and p53-dependent pathways[J].Toxicology & Applied Pharmacology,2012,262(1):80-90.
[27] 王健,侯惠民,夏怡然,等.毛萼乙素纳米混悬剂.II.对小鼠肝癌H22移植瘤的生长抑制作用[J].中国医药工业杂志,2012,43(3):194-197.
[28] 刘建平.生物药剂学与药物动力学[M].北京:人民卫生出版社,2000:263-270.
[29] Ludden T M.Nonlinear Pharmacokinetics[J].Clinical Pharmacokinetics,1992,20(6):429-446.
[30] Iwasaki K,Murayama N,Koizumi R,et al.Comparison of cytochrome P450 3A enzymes in cynomolgus monkeys and humans[J].Drug Metabolism & Pharmacokinetics,2010,25(4):388-391.
[31] Aueviriyavit S,Kobayashi K,Kan C.Species Differences in Mechanism-Based Inactivation of CYP3A in Humans,Rats and Mice[J].Drug Metabolism & Pharmacokinetics,2010,25(1):93-100.
[32] Iwasaki K,Murayama N,Koizumi R,et al.Comparison of cytochrome P450 3A enzymes in cynomolgus monkeys and humans[J].Drug Metabolism & Pharmacokinetics,2010,25(4):388-391.
[33] Usmani K A,Hodgson E,Rose R L.In vitro metabolism of carbofuran by human,mouse,and rat cytochrome P450 and interactions with chlorpyrifos,testosterone,and estradiol[J].Chemico-Biological Interactions,2004,150(3):221-232.
[34] Guo F,An T,Rein K S.The algal hepatoxoxin okadaic acid is a substrate for human cytochromes CYP3A4 and CYP3A5[J].Toxicon,2010,55(2):325-332.
[35] Tang J,Usmani K A,Hodgson E.In vitro metabolism of fipronil by human and rat cytochrome P450 and its interactions with testosterone and diazepam[J].Chemico-Biological Interactions,2004,147(3):319-329.