LC/DAD/MSD技术研究大鼠胆汁中盐酸非洛普的II相代谢产物

目的研究大鼠服药后胆汁中盐酸非洛普(DDPH)II相代谢产物.方法收集大鼠空白胆汁及给药后12h内的胆汁,以LC/DAD/MSD技术判断II相代谢产物峰位.以HPLC法制备II相代谢产物馏分并以β-葡糖醛酸酶水解,再进行分析;同时将与II相代谢产物相应的I相代谢产物对照品按相同条件进行分析对照.结果大鼠给药后胆汁色谱图中峰M₇,M₈和M₉为DDPH的II相代谢产物,它们的β-葡糖醛酸酶水解产物分别为M₃,M₂和M₁.结论β-1-O-{3,5-二甲基-4-[-2-甲基-2-(3,4-二甲氧基苯乙氨基)-乙氧基]-苯基}-葡糖醛酸(M₇),β-1-O-{2,4-二甲基-3-[2-甲基-2-(3,4-二甲氧基苯乙氨基)-乙氧基]-苯基}-葡糖醛酸(M₈)和β-1-O-{2-甲氧基-4-[1-甲基-2-(2,6-二甲基苯氧基)-乙氨基-乙基]-苯基}-葡糖醛酸(M₉)为大鼠ipDDPH后产生的II相代谢产物.     

LC-MS/MS及离子簇技术分析鉴定盐酸戊乙奎醚外消旋体在大鼠体内的代谢产物

目的研究盐酸戊乙奎醚外消旋体在大鼠体内的代谢产物。方法健康大鼠同时等量im盐酸戊乙奎醚外消旋体及其氘标盐酸戊乙奎醚,收集尿样并处理。用LC/MS/MS,GC-MS,FAB-MS及氘标离子簇示踪技术分析鉴定盐酸戊乙奎醚在大鼠体内的代谢产物。结果共检测到8个代谢产物,分别为原形环戊基上的单氧化产物(M1与M1^*)、原形环戊基上的单羟基化产物(M2与M2^*)、原形环戊基间位上的氧化羟基化产物(M3与M3^*)及原形环戊基与奎宁环上的羟基化产物(M4与M4^*)。其中,M1与M1^*,M2与M2^*,M3与M3^*及M4与M4^*互为异构体。结论 此法为临床合理用药及新抗胆碱能手性药物研究提供有价值的信息。     

5-取代-4-甲基-1,3-二氢-2H-咪唑-2-酮类化合物的合成及其强心作用

本文设计并合成了5-取代-4-甲基-1,3-二氢-2 H-咪唑-2-酮类化合物23个,其中19个尚未见报道。中间体5-(4-溴甲基苯甲酰基)-4-甲基-1,3-二氢-2H-咪唑-2-酮(Ⅴ)为一新化合物。在关键中间体4-甲基-1,3-二氢-2H-咪唑-2-酮(Ⅳ)的合成中,设计了由β-羟基丙胺一步氧化直接合成氨基丙酮的新方法。药理初筛表明:Ⅱ₁,Ⅱ₂,Ⅱ₄,Ⅲ₄,Ⅲ₆,Ⅲ₇,Ⅲ₈和RMI 82,249(1)具有不同程度的正性肌力作用,其中4个未知化合物Ⅱ₁,Ⅱ₂,Ⅱ₄和Ⅲ₄的最大效应高于已知对照物(1)。Ⅱ₄既增加心肌收缩力,又减慢心率,值得进一步研究。     

4-((4-((2-取代苯氧基)乙基)-1-哌嗪)-甲基)-1,2-二氢喹啉-2-酮类化合物的合成及体外α-受体拮抗活性

目的研究DDPH类似物1,2-二氢喹啉-2-酮类化合物的合成及其体外α-受体拮抗活性。方法通过酰化、溴代、环合和取代反应等合成目的物;推测了异常中间体3-溴-4-溴甲基-1,2-二氢喹啉-2-酮(5)和3-溴-4-甲基-1,2-二氢喹啉-2-酮(6)的生成机理;测定目的物的体外α-受体拮抗活性。 结果设计、合成了12个新化合物II_1～6和IV_1～6,其中6个目的物1,2-二氢喹啉-2-酮类(IV_1～6)的结构经IR,¹HNMR,MS和HRMS确证;IV₃,IV₄和IV₆对兔主动脉环抑制作用较明显。结论化合物IV₃,IV₄和IV₆显示了一定的抑制活性,值得进一步研究。     

人参皂苷Rb1在大鼠肠内菌代谢物吸收入血成分的研究

目的:研究口服人参皂苷Rb₁(G-Rb₁)被吸收入血的成分。方法:给大鼠igG-Rb₁500mg·kg^-1后,于不同时间采集粪、尿及血清样品,用电喷雾质谱(ESI-MS)检测吸收入血成分。结果:在血与尿中发现分子量为1108,946及784amu的代谢产物。经ESI-MS2级质谱分析,上述分子量的化合物分别为G-Rb₁,Rd和F₂。结论:G-Rb₁给大鼠ig后,原形及中间代谢产物Rd及F₂被吸收入血。     

拟人参皂苷F11在大鼠体内的药物代谢研究

目的探讨拟人参皂苷F11在大鼠体内的药物代谢产物及其过程.方法ip拟人参皂苷F₁₁后,应用TLC分析排泄物中的代谢产物,并利用制备薄层分离制备代谢产物,通过波谱解析(MS,¹HNMR,¹³CNMR,¹H-¹HCOSY)确定其结构.结果从粪便中分离鉴定了3种代谢产物,分别为拟人参皂苷R_T5,ocotillol和1个新的代谢产物F-3-1,并确定其结构为6-O-α-L-吡喃鼠李糖基(1-2)-β-D-吡喃葡糖基-(20S,23S,24R)-达玛-20(24)-环氧-3β,6α,12β,23,25-五醇(6-O-α-L-rhamnopyranosyl-(1-2)-β-D-glucopyranosyl-(20S,23S,24R)-dammar-20(24)-epoxy-3-β,6α,12β,23,25-pentanol).但在尿液和胆汁中并未发现任何代谢产物.结论拟人参皂苷F₁₁不被肝脏代谢,但胆汁排泄物可在肠道被代谢为水解和氧化产物.     

山莨菪碱对大鼠胸水中性白细胞花生四烯酸代谢的影响

本文研究了654-2对大鼠胸水中性白细胞代谢[1-¹⁴C]AA及内源性AA的影响。大鼠白细胞AA经5-LPO代谢途径形成的主要产物为LTB₄及5-HETE,经CO途径的主要产物为HHT及TXB₂。654-2对白细胞代谢[1-¹⁴C]AA无抑制作用,但显著减少白细胞从内源性AA形成的LTB₄,5-HETE,HHT及TXB₂。这种抑制作用与654-2呈剂量依赖关系。本实验结果表明,654-2抑制PG及LT的形成可能是影响了AA从胞膜的释放,而并非直接抑制CO及5-LPO。     

甲基莲心碱在大鼠肝脏中的代谢产物及其途径

大鼠灌胃给予甲基莲心碱 20 mg·kg^-1，采用液相色谱-串联质谱联用法对大鼠肝脏中的代谢产物进行分析；并建立肝微粒体温浴及NADPH再生体系，采用高效液相色谱-紫外检测法研究CYP450亚型的特异性抑制剂对甲基莲心碱体外代谢的影响。在正离子检测方式下，除甲基莲心碱外共检测到4种代谢产物M₁、M₂(主要代谢产物)、M₃和M₄。其中，M₂和M₄通过与对照品的色谱和质谱比对，确认为莲心碱和异莲心碱，而M₁ 和M₃可能为去甲基莲心碱和去甲基异莲心碱。CYP3A1的特异性抑制剂酮康唑和CYP2D1的特异性抑制剂奎尼丁均可抑制甲基莲心碱在肝微粒体温孵液中的代谢，其主要代谢产物莲心碱的生成抑制率分别为25.7%和80.5%。因此提示，甲基莲心碱在肝脏中的主要代谢途径是苄基和喹啉环上的甲氧基脱甲基化，其主要代谢物为莲心碱，CYP2D1和CYP3A1均参与了其生物转化。     

HPLC-MSn法鉴定葫芦巴碱及其在大鼠体内的主要代谢产物

目的建立快速灵敏的LC-MSⁿ检测葫芦巴碱及其在大鼠体内代谢物的分析方法。方法以葫芦巴碱对LC-MS²色谱及质谱条件进行优化，分析其电喷雾质谱的一级电离规律和多级质谱裂解规律，以此作为葫芦巴碱大鼠体内代谢物分析鉴定的依据。健康大鼠尾静脉注射8 mg·kg^-1葫芦巴碱，收集0～48 h的尿样，经C₁₈小柱固相萃取分离纯化后，直接采用LC-MSⁿ方法对尿样进行测定。结果根据生物体内药物代谢转化规律及母体药物的色谱-质谱行为规律，在尿样中鉴定出母药及其N-去甲基、N-去甲基环氧化产物，以及母药及其N-去甲基环氧化物的甘氨酸轭合物。结论本方法灵敏、快速、选择性高、专属性好，可用于葫芦巴碱的代谢产物研究。     

人参皂甙Rb1的肠内菌代谢

通过离体及整体实验观察了人和大鼠肠内菌对人参皂甙Rb₁（G-Rb₁）的代谢。方法：采用薄层色谱（TLC）和电喷雾质谱（ESI-MS）检测G-Rb₁及其代谢产物。结果：离体实验表明,G-Rb₁容易被大鼠和人消化道菌群代谢,随着代谢时间的延长,相继出现Rd, Rg₃/F₂, Rh₂/C-K和Ppd 4种代谢产物。给大鼠ig G-Rb₁ 500 mg.kg^-1后收集4 h和6 h粪,提取G-Rb₁的代谢产物,证明粪中存在Rd和Rg₃/F₂两种代谢产物。结论：G-Rb₁可被人和大鼠肠内菌代谢,其代谢模式为G-Rb₁→Rd→F₂→compound K（C-K）→20（S）protopanaxadiol（Ppd）。     

Lack of effect of spiramycin on cyclosporin pharmacokinetics.

1. The influence of spiramycin coadministration on cyclosporin pharmacokinetics was studied in five renal transplant patients. The plasma concentrations of cyclosporin were measured both by non-specific radioimmunoassay (RIA) and high-performance liquid chromatography (h.p.l.c.). 2. The kinetics of cyclosporin were followed before treatment, and after 1 day and then 2 weeks of oral treatment with spiramycin (3 X 10(6) iu, twice daily). The main pharmacokinetic parameters (the area under the plasma drug concentration-time curve, the maximum plasma drug concentration and the time to reach it) obtained both by RIA and h.p.l.c. were not modified by spiramycin cotreatment after 1 day, nor after 2 weeks of spiramycin administration. Therefore, the pharmacokinetics of cyclosporin (parent drug and parent drug plus metabolites) are not influenced by the coadministration of spiramycin macrolide at therapeutic dosage. 3. Spiramycin may be preferable to other macrolide antibiotics known to interact with cyclosporin such as erythromycin or josamycin.     

花椒毒酚抑制大鼠局灶性脑缺血/再灌注损伤后中性粒细胞浸润和脑水肿

目的研究花椒毒酚对大鼠局灶性脑缺血/再灌注损伤后中性粒细胞浸润、细胞粘附分子表达及脑水肿的影响。方法线栓法短暂阻塞大鼠大脑中动脉制备局灶性脑缺血(2 h)再/灌注(24 h)损伤模型,缺血后1 h和12 h两次腹腔注射花椒毒酚2.5、5和10 mg.kg-1,再灌注24 h,检测大鼠神经功能行为缺陷评分、脑水肿和脑梗死范围,分光光度法测定缺血区脑组织中Na+,K+-ATPase、Ca2+-ATPase和髓过氧化物酶(MPO)的活性,免疫组织化学染色测定大脑皮层细胞间粘附分子-1(ICAM-1)和E选-择素(E-selectin)的表达。结果花椒毒酚能改善大鼠脑缺血/再灌注后神经功能行为缺陷评分,减轻脑水肿和降低脑梗死范围,增强Na+,K+-ATPase和Ca2+-ATPase活性,降低脑组织中MPO的活性,抑制ICAM-1和E-selectin表达。结论花椒毒酚对脑缺血/再灌注损伤有保护作用,其机制可能与其抑制炎症反应和减轻脑水肿有关。     

Metabolism and pharmacokinetics of a dipeptidyl peptidase IV inhibitor in rats, dogs, and monkeys with selective carbamoyl glucuronidation of the primary amine in dogs.

The pharmacokinetics and metabolism of the l-threo isoleucine thiazolidide dipeptidyl peptidase IV inhibitor, di-[2S,3S]-2-amino-3-methyl-pentanoic-1,3-thiazolidine fumarate (ILT-threo) and its allo stereoisomer (ILT-allo) were evaluated in rats, dogs, and monkeys. Both compounds were well absorbed (>80%) in all species, and most of the dose (>60%) was recovered in urine. Metabolites identified in all species included a sulfoxide (M1), a sulfone (M2), and a carbamoyl glucuronide (M3). For both compounds, parent drug had moderate systemic clearance in rats and dogs ( approximately 20-35 ml/min/kg in both species) and lower clearance in monkeys ( approximately 6-9 ml/min/kg). In rats, M1 was present in systemic circulation in concentrations similar to that of parent drug, whereas in dogs and monkeys, exposures to M1 were higher than for parent drug. In dogs, exposures to the sulfoxide metabolite were approximately 2 to 3 times higher after administration of ILT-allo than after administration of ILT-threo. Carbamoyl glucuronidation was an important biotransformation pathway in dogs. Circulating levels of M3 were significant in the dog, and present only in trace levels in rats and monkeys. M3 could be produced in in vitro systems in a NaHCO3 buffer under a CO2-saturated atmosphere and in the presence of UDP-glucuronic acid and alamethicin.     

Disposition and metabolism of a new benzothiophene antiestrogen in rats,dogs and monkeys

1. A new benzothiophene-derived antiestrogen (LY156758) when orally administered was well absorbed in rats and monkeys while approx. 20% was absorbed in dogs.

2. In the rat the compound was subject to first-pass hepatic metabolism which led to low levels of parent drug in the systematic circulation together with a small amount as the glucuronide conjugate.

3. In monkeys the compound occurred primarily as the glucuronide conjugate of parent drug with very little circulating free drug.

4. The systemic bioavailability of free parent drug in plasma was 39% in rats, 17% in dogs and 5% in monkeys.

5. Excretion of the drug in rats and dogs was primarily via the bile. Approx. 1% of the dose was excreted in the urine of rats and dogs after oral dosing. In rats, at least 50% of an oral dose was excreted in bile as the glucuronide conjugate of parent drug.     

A sensitive analytical method for the determination of a novel lipoxygenase inhibitor, 4-bromo-2,7-dimethoxy-3H-phenothiazin-3-one, in plasma using reductive electrochemical detection

A reversed-phase high-performance liquid chromatographic assay using electrochemical detection in the reductive mode has been developed for the analysis of 4-bromo-2,7-dimethoxy-3H-phenothiazin-3-one (1) in plasma to determine drug absorption. Free drug in plasma in concentrations as little as 0.25 ng/mL can be estimated with a mean coefficient of variation (CV) of 6.3 +/- 2.6%. Metabolites which can be converted to the parent drug by acid hydrolysis can be quantified in concentrations of 10 ng/mL or more, with a mean CV of 4.3 +/- 1.9%. To test the procedure, plasma was obtained from dogs receiving 14C-labeled 1. After acid hydrolysis of plasma, the electrochemical assay for parent drug showed good agreement with the radioactive equivalents in plasma, suggesting that parent drug and metabolites can be satisfactorily analyzed by this procedure.     

溴泰君在大鼠的组织分布和排泄

目的研究溴泰君(W198)在大鼠的组织分布和排泄，为临床试验提供依据。方法用HPLC紫外检测方法测定大鼠iv W198后生物样品中的药物含量。结果大鼠iv W198 20 mg·kg^-1后组织的药物含量远高于相同时间的血清药物浓度。药物主要分布在肺，其次在肾、心、肝等组织,大部分组织的药物含量于给药后0.25 h最高，给药后2 h显著下降，至24 h均缓慢下降；大鼠iv W198 20 mg·kg^-1后从尿、粪(0-96 h)和胆汁(0-24 h)中排泄的原型药物分别占给药总量的0.150%，2.1%和0.063%。结论W198主要分布于肺组织中。从尿、粪和胆汁中排泄的原型药物很少。     

盐酸甲唑醇的药代动力学及片剂与胶囊的生物利用度比较

犬po盐酸甲唑醇(methimidol hydrochlo-ride)片剂或胶囊后,用分光光度计测定了不同时间的血药浓度,用3P87电子计算机程序计算了药代动力学参数,对两种剂型的生物利用度进行了比较。结果表明该药在犬体内以双指数函数衰减,即符合二室开放模型,胶囊与片剂的末相半衰期分别为6.9 h和6.1 h,片剂的相对生物利用度为胶囊的80.6％。     

Inhibitors of human immunodeficiency virus type 1 (HIV-1) attachment 6. Preclinical and human pharmacokinetic profiling of BMS-663749, a phosphonooxymethyl prodrug of the HIV-1 attachment inhibitor 2-(4-benzoyl-1-piperazinyl)-1-(4,7-dimethoxy-1H-pyrrolo[2,3-c]pyridin-3-yl)-2-oxoethanone (BMS-488043)

BMS-663749, a phosphonooxymethyl prodrug 4 of the HIV-1 attachment inhibitor 2-(4-benzoyl-1-piperazinyl)-1-(4,7-dimethoxy-1H-pyrrolo[2,3-c]pyridin-3-yl)-2-oxoethanone (BMS-488043) (2) was prepared and profiled in a variety of preclinical in vitro and in vivo models designed to assess its ability to deliver parent drug following oral administration. The data showed that prodrug 4 had excellent potential to significantly reduce dissolution rate-limited absorption following oral dosing in humans. Clinical studies in normal healthy subjects confirmed the potential of 4, revealing that the prodrug significantly increased both the AUC and C(max) of 2 compared to a solid capsule formulation containing the parent drug upon dose escalation. These data provided guidance for further efforts to obtain an effective HIV-1 attachment inhibitor.     

Pharmacokinetics of the anti-AIDS drug 2',3'-dideoxyinosine in the rat

The pharmacokinetics of 2',3'-dideoxyinosine (ddlno) was examined in the male Fischer 344 rat using reversed phase HPLC with UV and radiochemical detection. Following iv doses of 12, 25, and 100 mg/kg, the parent drug was rapidly eliminated from plasma (mean residence time, 6.3 min; systemic clearance, 64 ml/min/kg). The mean terminal elimination half-life was 28 min and the volume of distribution at steady state was 0.39 liter/kg. Orally administered [3H]ddlno (100 mg/kg) was not significantly bioavailable in the rat, with only 8-11% of the dose absorbed as the parent compound over a 2 hr period. Peak plasma drug concentrations occurred 10 to 20 min following oral administration. Solution stability data suggested that ddlno was unstable at gastric pH (pH 1, t1/2 less than 30 sec). However, elevation of gastric pH with sodium bicarbonate prior to oral administration of ddlno was not effective in increasing the bioavailability of the parent drug. The drug and its metabolic products were extensively distributed into rat tissue 48 hr after iv or oral administration. Following iv infusion of [3H]ddlno to steady state conditions, the highest tissue-to-plasma ratios of radioactivity were found in the kidney (2.2), liver (1.7), and spleen (1.5). Renal clearance accounted for 99% of the eliminated dose. Liver perfusion studies showed that ddlno was not subject to significant hepatic clearance (less than 10%) and that the metabolism was not inducible with phenobarbital.