摘要
海洋硫酸多糖916是已由国家药品监督管理局批准的进入临床试验的抗动脉粥样硬化的?类新药。临床上治疗心血管病症多为联合用药,在开发新药过程中若希望与其它药物共用,应考虑药物的相互作用。洛伐他汀(lovastatin,以下简称LV),为羟甲戊二酰辅酶A(HMC-CoA)还原酶抑制药,抑制内源性胆固醇的合成,与916同为抗动脉粥样硬化药物。目前国内有关药物动力学方面的相互作用报道较少,本文选择洛伐他汀作为模型药物,916作为合用成分,以洛伐他汀的大鼠体内体外药物动力学为基础,研究合用916后LV的大鼠体内药物动力学参数的变化,并研究了体外肝微粒体中916对LV的代谢的影响。
1.本文确立了用于生物样品分析并经改良后的高效液相色谱条件,即选用XTerra? MS C18色谱柱(150×2.1mm, 5μm),乙腈和水(63:37,v/v)作为流动相,流速为0.2m1/min,柱温设在35℃,测定血浆、大鼠排泄物及肝微粒体中LV的含量。LV在0.01～50mg·L-1范围内呈线性关系,相关系数均大于0.99;血浆和尿液中LV定量限约为0.01mg·L-1;血浆中的LV的提取回收率在80%以上;LV的日内差、日间差均小于15%。此方法准确可靠、灵敏度高、专属性强,很好地满足了本文中的各项样品分析的要求。
2.采用灌胃给药法,分别研究了LV在雄性和雌性大鼠体内的药动力学及合用916后LV的动力学性质。合用916后雄性大鼠体内LV的t1/2由4.2±0.8h变为2.5±0.5h,AUC0→∞由763.0±58.0h·μg·L-1变为3678.7±1445.4h·μg·L-1,Cmax由173.2±32.4μg·L-1变为1333.1±211.4μg·L-1其在统计学上均有显著性意义(P<0.05);雌性大鼠体内LV的显著性变化有t1/2由5.9±2.1h变为3.6±0.5h,AUC0→10由397.0±95.5h·μg·L-1变为561.56±167.4h·μg·L-1。
3.采用代谢笼法,研究了LV在雄性大鼠尿液和粪便中的排泄。结果发现LV原形药的74h累积排泄率在尿中为5.95%,粪便中为46.26%,给药74h后,尿中已无原形药物出现;经t-检验,单用LV与合用916组的0~10h和0~24h段原形药物的尿液累积排泄量减少,并有有统计学显著差异(P < 0.05),但粪便中的累积排泄率无显著差异(P>0.05)。
4.采用体外肝微粒体法,研究了LV在雄性大鼠肝微粒体中的代谢。结果表明LV在混合酶体系中在5-60min呈线性代谢,在60-90min代谢缓慢,916与LV共同孵育对LV的代谢的影响无显著性。
5.采用体外探针法,分别研究了916对探针药物咖啡因、氨苯砜和氯唑沙宗在雄性和雌性大鼠肝微粒体中代谢的影响。结果表明916对大鼠肝微粒体中三种CYP亚型酶(CYP1A2、CYP2E1、CYP3A)无诱导或抑制作用。
Marine Sulfated Polysaccharide 916 was authorized to go into clinical study further by National Drug and Foods Administration at the name of I level new drug with activity of anti-atherosclerosis . Drug combination is usually used for cardiovascular disorder clinically, to combine with other drug, drug interaction must be considered in development of new drugs. Lovastatin (LV) is the inhibitor of HMC-CoA and can inhibit synthesis of endogenous cholesterol and can heal atherosclerosis like 916. There are few domestic reports about pharmacokinetic interaction. This paper chose LV as model drug and 916 as combined drug to study the diversity of pharmacokinetics parameter of LV in rats after a single oral does of LV alone and with 916 together. This paper also investigated the influence of metabolism of LV in liver- microsome caused by 916.
1.In this paper,an analytical method in biological sample was developed by HPLC: chromatography was carried out using a XTerra? MS C18 column (150×2.1mm, 5μm) with a mobile phase consisting of acetonitrile-water(63:37, v/v) at a flow rate of 0.2 ml/min and 35℃as temperature of column. The calibration plots of LV in biological sample were linear over a range from 0.01 to50 mgL-1, R2 were all exceed 0.99; LOQ of LV in plasma and urine were about 0.01 mgL-1; extract recovery of LV in plasma was more than 80%; the analytical precision of intra-day and inter-day were less than 15%. This analytical method is simple, high sensitive and high reduplicative enough to be used in this pharmacokinetic study.
2. To investigate the pharmacokinetics of LV in rats, LV was administered alone or in combination with 916. When in combination with 916, the plasma concentration of LV in male rats, t1/2 was changed from 4.2±0.8h to 2.5±0.5h, AUC0→∞was changed from 763.0±58.0h·μg·L-1 to 3678.7±1445.4h·μg·L-1,Cmax was changed from 173.2±32.4μg·L-1 to 1333.1±211.4μg·L-1, the changes were all conspicuous (P<0.05); in female rats, t1/2 was changed from 5.9±2.1h to 3.6±0.5h, AUC0→10 was changed from 397.0±95.5h·μg·L-1 to 561.56±167.37h·μg·L-1(P<0.05).
3. To investigate the elimination of LV from male rats urine and fecal by after a single dose of LV, metabolism cages method was used. The results showed that the accumulated excretive ratio of unchanged LV was 5.95% from urine,46.26% from fecal for 74h after dosing,and LV almost was indiscoverable in urine after 74h after dosing .When co-administered with 916,the elimination of LV from urine during 0~10h and 0~24h decreased significantly (P<0.05), but there were no significant changes by fecal(P>0.05).
4. To investigate the metabolism of LV from male rat liver-microsome after a single dose of LV, metabolism of liver-microsome in vitro method was used. The result showed the metabolism velocity of LV was linear over a range from 0 to 60 min, the metabolic velocity of LV decreased over 60-90min. There were no significant changes when 916 was co-incubated.
5. To investigate the influence of metabolism of caffeine, dapsone and chlorzoxazone in liver-microsome of rats caused by 916, P450 probe assays was used. The results show 916 have neither notable induced nor inhibitive effect on CYP1A2, CYP2E1, CYP3A enzymes.
引文
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