附杞固本膏在中国健康受试者中的随机、双盲、安慰剂对照、连续给药的药代动力学及代谢组学分析  

Pharmacokinetic and Metabolomic Analysis of Randomized,Double-blind,Placebo-controlled,Continuous Administration of Fuqi Gubengao in Healthy Chinese Subjects

作　　者：刘峻宇 曹唯仪[2] 孙敏 陆芳[2] 杨巧宁[2] 张菀桐 曲华[2] 王淑阁[2] 李睿[2] 高蕊[2] LIU Junyu;CAO Weiyi;SUN Min;LU Fang;YANG Qiaoning;ZHANG Wantong;QU Hua;WANG Shuge;LI Rui;GAO Rui(Yunnan Province Company Key Laboratory for Traditional Chinese Medicine(TCM)and Ethnic Drug of New Drug Creation,Yunnan Institute of Materia Medica,Kunming 650111,China;Key Laboratory for Clinical Research and Evaluation of TCM of National Medical Products Administration,National Clinical Research Center for Chinese Medicine Cardiology,Xiyuan Hospital of China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]云南省药物研究所,云南省中药和民族药新药创制企业重点实验室,昆明650111 [2]中国中医科学院西苑医院,国家中医心血管病临床医学研究中心,中药临床研究与评价重点实验室,北京100091

出　　处：《中国实验方剂学杂志》2024年第16期160-169,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中国中医科学院科技创新工程项目(CI2021A04703,CI2021A04701);中央级公益性科研院所基本科研业务费专项(ZZ15-YQ-019);云南省科技厅“科技人才与平台计划”项目(202205AD160044)。

摘　　要：目的:评价健康受试者对附杞固本膏的耐受程度,探索附杞固本膏主要活性成分的人体药代动力学特征及治疗肾阳虚证的作用机制。方法:采用单中心、随机、双盲、连续给药、安慰剂对照设计。共纳入健康受试者24例,设置75 g和100 g 2个剂量组,每剂量组12例受试者按2∶1随机分为试验组和安慰剂组,各组按剂量给予药物或安慰剂,连续给药14 d,期间记录受试者体征以进行安全性评估。按方案规定的时间点采集生物样本,通过超高效液相色谱-三重四级杆串联质谱法(UPLCQqQ-MS/MS)检测生物样本中乌头碱、新乌头碱、次乌头碱、苯甲酰乌头原碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱的血药浓度,通过非房室模型计算药代动力学参数。以超高效液相色谱-四极杆飞行时间质谱法(UPLC-Q-TOF-MS)检测血浆和尿液样本中内源代谢物的相对含量,使用EZinfo 3.0软件进行主成分分析(PCA)、偏最小二乘法-判别分析(PLS-DA)和正交偏最小二乘法-判别分析(OPLS-DA),利用变量重要性投影(VIP)值>1和t检验P<0.05筛选差异代谢物,并利用代谢物分析平台MetaboAnalyst 6.0进行通路富集分析。结果:在安全性评价中,未观察到不良事件随着剂量的增加而明显增加的趋势。药代动力学结果显示,乌头碱、新乌头碱、次乌头碱的血浆浓度多数低于定量下限,苯甲酰乌头原碱首次给药后71%的血药浓度低于定量下限。75 g组和100 g组苯甲酰新乌头原碱在连续给药12 d后血药浓度达到稳态,末次给药后体内药物浓度及整体暴露较首次给药有所增加,但未出现安全性相关的药物蓄积。血中鉴定出附杞固本膏诱导的差异代谢物45个,包括酰基肉碱11种、脂肪酸10种、氨基酸4种、脂质20种,富集到α-亚麻酸与亚油酸代谢、支链脂肪酸氧化、苯丙氨酸和酪氨酸代谢、脂肪酸生物合成等通路。尿中鉴定出差异代谢物22种,主要包括氨Objective:To evaluate the tolerance of healthy subjects to Fuqi Gubengao,and to explore the pharmacokinetics of its major active components and the potential mechanisms in treating syndrome of deficiency of kidney Yang.Method:A Single-center,randomized,double-blind,continuous administration,placebo-controlled trial was designed.A total of 24 healthy subjects were enrolled in 2 dosage groups(75 g and 100 g),with each group consisting of 12 participants randomized in a ratio of 2∶1 to either the experimental and placebo groups,and each group was given the drug or the placebo at the dose for 14 d,the physical signs of subjects were recorded for safety assessment.Biological samples were collected at the specified time points according to the trial protocol.The concentrations of 6 aconite alkaloids(aconitine,mesaconitine,hypaconitine,benzoylaconine,benzoylmesaconine and benzoylhypacoitine)in the biological samples were determined using ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-QqQMS/MS),and pharmacokinetic parameters were calculated by non-compartment model.The relative contents of endogenous metabolites in plasma and urine samples were measured by UPLC-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS).Principal component analysis(PCA),partial least squares-discriminant analysis(PLS-DA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were performed by EZinfo 3.0 software,differential metabolites were screened with variable importance in the projection(VIP)value>1 and P<0.05,and pathway enrichment analysis was conducted using MetaboAnalyst 6.0 platform.Result:In the safety evaluation,no trend of significant increase in adverse events with increasing dose was observed.Pharmacokinetic results showed that the plasma concentrations of aconitine,mesaconitine and hypaconitine were mostly below the limit of quantification,and 71%of the plasma concentration of benzoylaconine was below the limit of quantification after the initial administration.The

关 键 词：附杞固本膏 Ⅰ期临床研究 药代动力学 代谢组学 安全性 耐受性 

分 类 号：R22[医药卫生—中医基础理论] R969.1[医药卫生—中医学] R28R24