5-Fu多级前体药物的设计合成与体内抗肿瘤作用

目的:设计合成5-Fu的多级前体药物,并进行小鼠体内抗肿瘤作用研究。方法:在5-Fu的N_1和N_3位引入邻甲苯甲酰基,分别合成氟尿嘧啶的邻甲苯甲酰基的单取代(TFu)和双取代(DTFu)前体药物,并考察目标化合物对小鼠S_(180)实体瘤和肝癌实体瘤的抑瘤作用。结果:在相同的给药剂量下(按100 mg·kg~(-1)给药时)DTFu和TFu对的小鼠S_(180)实体瘤的抑瘤率分别为55．88%和57．84%,优于对照药FT-207(抑瘤率为44．12%)。对肝癌实体瘤实验,DTFu抑瘤率与剂量成相关性,50,100和200 mg·kg~(-1)时的抑瘤率分别为37．74%,58．14%和88．74%。DTFu对小鼠灌胃给药的LD_(50)为3 165．77 mg·kg~(-1)。结论:TFu和DTFu具较强抗肿瘤活性且毒性较低。     

阿托氟尿嘧啶体内代谢物的检测

导数光谱法测定小鼠口服A-OT-Fu(1-乙酰-3-邻甲苯甲酰-5-氟尿嘧啶,阿托氟尿嘧啶)淀粉液后,体内的主要代谢产物TFu(3-邻甲苯甲酰-5-氟尿嘧啶)和5-Fu的血浓度。两者达到最高血药浓度的时间分别为1和2h。     

用四阶导数光谱法测定A-OT-Fu小鼠代谢物的血药浓度

本文采用四阶导数光谱测定了小鼠口服A-OT-Fu淀粉液后的主要代谢产物TFu和5-Fu的血药浓度—时间曲线。当TFu和5-Fu浓度为5～50μg/ml时,浓度与252nm和289 nm对零轴间的振幅有良好的线性关系。在本测试条件下TFu和5-Fu的回收率约95%以上,实验结果表明,TFu和5-Fu达到最高血药浓度的时间分别为1和2 h。     

用四阶导数光谱法测定A-OT-Fu小鼠代谢物的血药浓度

本文采用四阶导数光谱测定了小鼠口服A-OT-Fu淀粉液后的主要代谢产物TFu和5-Fu的血药浓度—时间曲线。当TFu和5-Fu浓度为5～50μg/ml时,浓度与252nm和289 nm对零轴间的振幅有良好的线性关系。在本测试条件下TFu和5-Fu的回收率约95%以上,实验结果表明,TFu和5-Fu达到最高血药浓度的时间分别为1和2 h。     

反相高效液相色谱法测定癌特灵在小鼠体内的主要代谢产物的血药浓度

本文报道用反相高效液相色谱法,在μBondapak C_(18)柱(30cm×3.8mm)上,以甲醇:水(4:6)为流动相,254nm处检测。以呋氟尿嘧啶为内标,测定了癌特灵一次口服给药后在小鼠体内主要代谢产物3-邻甲苯甲酸基-5-氟尿嘧啶(TFU)和氟尿嘧啶的血药浓度。操作方法简便,灵敏度高,重现性较好,符合临床药浓监测的要求,根据测定结果,TFU在小鼠体内的药物动力学过程符合二室开放模型,清除速率为一级。     

广金钱草黄酮成分在肾结石大鼠中的代谢研究

目的 利用超高效液相色谱串联电喷雾飞行时间质谱（UPLC-Q-TOF-MS/MS）分析广金钱草黄酮成分在肾结石大鼠中的代谢途径和代谢产物。方法 建立SD大鼠肾结石造模,灌胃给予肾结石大鼠广金钱草总黄酮,采用UPLC-Q-TOF-MS/MS分析黄酮成分在肾结石大鼠模型的血浆、尿液和粪便中的代谢变化,通过TOF/MS得到的准确相对分子质量,多级质谱裂解信息,结合对照品及相关文献报道对黄酮成分的裂解规律总结结果对比鉴定,对血浆、尿液和粪便中总黄酮成分和代谢产物进行鉴定。结果 广金钱草总黄酮给药后,在大鼠血浆中鉴定出5种原型成分和3种夏佛塔苷的同分异构体,16种以血浆中原型成分代谢得到的产物;从尿液中鉴定出12种原型成分和21种以尿液中原型成分代谢得到的产物;从粪便中鉴定10种原型成分和43种以粪便中原型成分代谢得到的产物。结论 阐述了广金钱草总黄酮在大鼠体内的代谢变化情况,为该药材今后的药效物质基础研究提供了理论参考。     

穿心莲内酯磺化物E在大鼠体内的代谢及代谢动力学研究

目的:对穿心莲内酯磺化物E在大鼠体内的代谢及代谢动力学进行研究。方法:大鼠分别口服、静注穿心莲内酯磺化物E后,采用LC-MS/MS法测定血浆中原型药物浓度,并估算其相应的药代动力学参数。采用LC-MS/MS法分析鉴定给药后大鼠血浆、尿液、胆汁和粪便中的代谢物。结果:口服给药后,穿心莲内酯磺化物E在大鼠体内吸收较为迅速,但绝对生物利用度仅为0.16%,并迅速从体内消除。进一步的代谢研究发现,静注和口服给药后在大鼠体内的主要代谢物分别为脱水产物,谷胱甘肽结合物及葡萄糖醛酸结合物,其在血浆中的主要存在形式为原型药物和脱水产物。结论:口服给药后,穿心莲内酯磺化物E以原型药物的形式吸收进入体内,并在体内进一步代谢。上述研究结果提示对穿心莲内酯C-19位的羟基进行磺化虽然提高了其水溶性,但降低了其口服生物利用度。     

液相色谱-串联质谱法分析抗肿瘤新药乙烷硒啉在大鼠体内的代谢产物

本研究对抗肿瘤新药1,2-[二(1,2-苯并异硒唑-3(2H)-酮)]乙烷(乙烷硒啉,BBSKE)在大鼠体内的代谢产物进行鉴定。在灌胃给予大鼠单剂量乙烷硒啉200mg·kg-1后,采用液相色谱-串联质谱法(LC-MSn)对大鼠尿液、粪样、胆汁和血浆中的代谢产物进行检测,通过全扫描和选择离子扫描,以及根据多级质谱裂解规律对代谢物的结构进行分析。研究发现在大鼠尿样、粪样、胆汁和血浆中检测到3种Ⅰ相代谢产物和1种Ⅱ相代谢产物,其代谢途径分别为氧化、甲基化、硫甲基化和葡萄糖醛酸化反应,提示乙烷硒啉在大鼠体内的代谢方式可能是通过氧化、甲基化及葡萄糖醛酸化反应形成代谢产物。     

UPLC-Q-TOF/MS法分析岩大戟内酯B在大鼠体内的代谢产物

目的:分析岩大戟内酯B在大鼠体内的代谢产物,预测其代谢途径。方法:将大鼠随机分为空白组(灌胃0.5%羧甲基纤维素钠溶液)和给药组(灌胃岩大戟内酯B,100 mg/kg),每组8只。分别收集给药后0~12、>12~24、>24~36 h的粪便,给药后0~2、>2~8、>8~12、>12~24、>24~36、>36~48 h的尿液以及给药后1、2、8、12、24、36 h的血液样品,分别以超声提取法、固相萃取法、蛋白沉淀法处理后,采用超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF/MS)联用技术和Analyst?TF 1.7.1、PeakView?2.2等软件联合分析、鉴定各样品中的代谢产物。结果与结论:从大鼠粪便中共检测到原型药物和7个代谢产物,从尿液和血液样品中分别检测到1、2个代谢产物。岩大戟内酯B灌胃后在大鼠体内经Ⅰ相的开环、脱水、氧化、脱氧、加氢反应等途径代谢;未检测到Ⅱ相代谢产物。     

20(R)-25-OCH_3-PPD在大鼠体内代谢产物的鉴定

目的鉴定抗肿瘤药物20(R)-25-OCH3-PPD在大鼠体内的代谢产物。方法大鼠单次灌胃给予药物20(R)-25-OCH3-PPD(40 mg·kg-1),收集粪便、尿液及血浆。采用LC-MS/MS法,以多反应监测(MRM)和二级全扫描质谱(Full Scan MS2)方式,对大鼠粪便、尿液及血浆中代谢产物进行分析和鉴定。结果在粪中检测到原形药物及8个代谢物,在血浆中检测到原形药物和1个去甲基代谢物,在尿液中没有检测到相关代谢物。结论 20(R)-25-OCH3-PPD在大鼠体内代谢广泛。     

人参皂苷Rg1 总被引：26，自引：1，他引：25

王毅  刘铁汉  王巍  王本祥 《药学学报》2000,35(4):284-288

Influence of the metabolic profile on the in vivo antioxidant activity of quercetin under a low dosage oral regimen in rats

BACKGROUND AND PURPOSE: Flavonoids are known to possess a broad set of pharmacological effects, some of which have been attributed to their antioxidant properties and, more recently, to cell signalling modulation. Nevertheless, flavonoids are extensively metabolized and their metabolites are the potential bioactive forms in vivo. Therefore, a first and crucial step to understand the mechanisms underlying potential health benefits of flavonoids is knowledge of their metabolites and their biological activities.EXPERIMENTAL APPROACH: To approximate a human dietary pattern of intake of flavonoids, regular rat chow was supplemented with 0.02% quercetin and fed to Sprague-Dawley rats over 3 weeks. Plasma samples were analysed by HPLC and electrospray tandem mass spectrometry, and plasma antioxidant capacity was measured by the 2,2'-azino-bis(3-ethylbenzothiazoline sulphonate) assay. KEY RESULTS: Major metabolites were 3'-methylquercetin (isorhamnetin) glucuronide sulphate conjugates, the most plausible conjugation positions being at the 3-, 5- and 7-hydroxyl positions. Isorhamnetin conjugates are methylated at the 3'-OH position, which decreases the high antioxidant activity of quercetin and its metabolites and their contribution to plasma antioxidant potential.CONCLUSIONS AND IMPLICATIONS: This metabolic pattern differs from that observed after a single high-dose administration, where the major metabolites were quercetin conjugates at 5- and 7-hydroxyl positions and a significantly increased plasma antioxidant activity was observed. These data show altogether that the different metabolic patterns obtained under a prolonged low-dosage regimen or after a single high dose, crucially affected the antioxidant potential of plasma in treated animals. Our data also allow for the establishment of structure-antioxidant activity relationships for quercetin metabolites.     

Simultaneous high-performance liquid chromatography assay of two metabolites of 6-chloro-2-pyridylmethyl nitrate, a new antianginal drug, and its application to a pharmacokinetic study in rats

An HPLC technique has been developed for the simultaneous determination of 6-chloro-2-pyridine methanol (5) and 6-chloro-2-pyridinecarboxylic acid (3), two metabolites of 6-chloro-2-pyridylmethyl nitrate, a new antianginal drug, in rat plasma. The plasma sample was placed on a Bond Elut C18 column and the compounds were eluted with 250 microL of the mobile phase. A 50-150-microL aliquot was injected onto the HPLC column. No interfering substances were observed in the plasma of a normal rat. The calibration curves for both 5 and 3 were linear from 0.1 to 50 micrograms/mL. However, the quantitative detection of the two additional metabolites, N-(6-chloro-2-pyridylcarbonyl)-glycine (1) and N-acetyl-S-(6-chloro-2-pyridylmethyl)-L-cysteine (2) was not satisfactory. The metabolic pathway of 6-chloro-2-pyridylmethyl nitrate in vivo was studied in male rats which were dosed separately with 6-chloro-2-pyridylmethyl nitrate, 5, and 3. After intravenous injection of 6-chloro-2-pyridylmethyl nitrate, the unchanged drug was determined by a GC method that was also developed by us and reported in another paper. 6-Chloro-2-pyridylmethyl nitrate was rapidly metabolized, and the metabolites were detected as early as 1 min after 6-chloro-2-pyridylmethyl nitrate administration. The parent compound then declined rapidly and 3 formation was confirmed in the rat plasma. 6-Chloro-2-pyridylmethyl nitrate is extensively metabolized to 3 via 5 formation, and 3 is the main metabolite in rat plasma.     

In vivo metabolism of [14C]ruboxistaurin in dogs, mice, and rats following oral administration and the structure determination of its metabolites by liquid chromatography/mass spectrometry and NMR spectroscopy.

Ruboxistaurin (LY333531), a potent and isoform-selective protein kinase C beta inhibitor, is currently undergoing clinical trials as a therapeutic agent for the treatment of diabetic microvascular complications. The present study describes the disposition and metabolism of [14C]ruboxistaurin following administration of an oral dose to dogs, mice, and rats. The study revealed that ruboxistaurin was highly metabolized in all species. Furthermore, the results from the bile duct-cannulated study revealed that ruboxistaurin was well absorbed in rats. The primary route of excretion of ruboxistaurin and its metabolites was through feces in all species. The major metabolite detected consistently in all matrices for all species was the N-desmethyl metabolite 1, with the exception of rat bile, in which hydroxy N-desmethyl metabolite 5 was detected as the major metabolite. Other significant metabolites detected in dog plasma were 2, 3, 5, and 6 and in mouse plasma 2, 5, and 19. The structures of the metabolites were proposed by tandem mass spectrometry with the exception of 1, 2, 3, 5, and 6, which were additionally confirmed either by direct comparison with authentic standards or by nuclear magnetic resonance spectroscopy. To assist identification by nuclear magnetic resonance spectroscopy, metabolites 3 and 5 were produced via biotransformation using recombinant human CYP2D6 and, likewise, metabolite 6 and compound 4 (regioisomer of 3 which did not correlate to metabolites found in vivo) were produced using a microbe, Mortierella zonata. The unambiguous identification of metabolites enabled the proposal of clear metabolic pathways of ruboxistaurin in dogs, mice, and rats.     

Pharmacokinetics of bupropion and metabolites in plasma and brain of rats, mice, and guinea pigs

Recent reports indicate that bupropion, a novel non-tricyclic antidepressant, is metabolized differently in certain species of animals. To further define the disposition of bupropion, a study was done involving three species, the rat, mouse, and guinea pig, as animal models to evaluate bupropion metabolism. The pharmacokinetic profiles of bupropion and its major basic metabolites, BW 306U and BW A494U, were determined following the ip administration of 40 mg/kg bupropion to these animals. Pharmacokinetic profiles of the parent drug and metabolites from plasma and brain samples were obtained using a liquid chromatographic procedure. Further investigation of the reduced bupropion metabolite BW A494U was carried out by the ip administration of this metabolite to these animals and assaying the plasma and brain samples 90 min after dosing. Analysis of the pharmacokinetic data revealed that the rat quickly metabolized bupropion, but no basic metabolites accumulated. The mouse metabolized bupropion predominantly to BW 306U, whereas the guinea pig converted bupropion to reduced bupropion (BW A494U) as well as BW 306U. Brain/plasma ratios of bupropion among these animals did not vary significantly. However, both metabolites showed dramatic differences in their brain/plasma ratios among these species. When reduced bupropion (BW A494U) was injected, almost 3% of the plasma concentration of BW A494U was determined to be bupropion in the rat. Lesser amounts were converted in the mouse and guinea pig. Therefore, we have demonstrated that distinct differences exist in the metabolism of bupropion in various species of animals. The guinea pig, when compared to the rat or mouse, appears to constitute a model that most closely resembles that of human bupropion metabolism.     

Metabolic switching of BILR 355 in the presence of ritonavir. I. Identifying an unexpected disproportionate human metabolite

11-Ethyl-5,11-dihydro-5-methyl-8-[2-[(1-oxido-4-quinolinyl)oxy] ethyl]-6H-dipyrido[3,2-b:2',3'-e][1,4]diazepin-6-one (BILR 355) is an inhibitor of the human immunodeficiency virus-1. BILR 355 exhibited a nonlinear pharmacokinetic profile and low exposure after oral administration to humans. This article describes the in vitro metabolism of BILR 355, which is correlated with the in vivo nonlinearity findings. Our in vitro studies had demonstrated that BILR 355 was extensively metabolized by cytochrome P450 3A. Thus, BILR 355 was concomitantly administered with ritonavir (RTV) in an attempt to boost systemic exposure, which did occur in humans. In addition, the expectation was that the overall metabolism of BILR 355 would be decreased with concomitant administration of RTV. Subsequent metabolite profiling was performed using human plasma samples obtained from clinical phase Ib studies with concomitant administration of BILR 355 and RTV. A total of 18 metabolites was observed. Their structures were proposed on the basis of high-performance liquid chromatography-tandem mass spectrometry technologies, and 10 metabolites were confirmed by comparison with synthetic standards. We were surprised to find that a disproportionate human metabolite, BILR 516, was uncovered during this metabolite profiling study and pharmacokinetic analysis of BILR 516 showed that it had a longer half-life and higher exposure than the parent compound at steady state. Of interest, BILR 516 was not detected in human plasma when BILR 355 was administered alone. Therefore, whereas RTV boosted the exposure of BILR 355, it resulted in a significant metabolic switching of BILR 355. Overall, this article demonstrates an unusual example of metabolic switching and raises concern about the consequence of metabolic switching during drug development.     

Metabolic pathways and pharmacokinetics of BOF-4272, a sulfoxide-containing drug, in the dog: in vivo and in vitro studies

BOF-4272, (+/-)-8-(3-methoxy-4- phenylsulfinylphenyl)pyrazolo[1,5-a]-1,3,5-triazine-4(1H)-one, is a new drug intended for the treatment of hyperuricemia. This report describes the detailed metabolic pathways of BOF-4272 in the dog. The metabolic pathways were investigated using the metabolites found in plasma, urine, and feces after intravenous or oral administration of BOF-4272, as well as the metabolites found in the liver S9 incubation mixture after the addition of BOF-4272 or BOF-4269. BOF-4269 (the sulfide metabolite of BOF-4272) was the only metabolite detected in plasma and feces after the intravenous or oral administration of BOF-4272. BOF-4269 was detected in dog plasma after a lag time following the oral administration of BOF-4272, and the Cmax and AUC0-t of BOF-4269 were higher in fed dogs than in fasted dogs after the oral administration of BOF-4272. A small amount of BOF-4269 was detected in dog plasma immediately after the intravenous administration of BOF-4272. Only BOF-4276 (the sulfone metabolite of BOF-4272) was detected in the S9 incubation mixture after the addition of BOF-4272. Mainly BOF-4272 was detected and small amounts of BOF-4276 and M-1 (the hydroxy metabolite of BOF-4269) were detected in the S9 incubation mixture after the addition of BOF-4269. These findings suggest that BOF-4272 is mainly metabolized to BOF-4269 by the intestinal flora in dogs, whereas little of this drug is metabolized to BOF-4269 in the dog liver. In conclusion, this work has allowed us to formulate the proposed metabolic pathways of BOF-4272 in the dog.     

Metabolic fate of 1-hexylcarbamoyl-5-fluorouracil after oral administration in mice

1. The metabolic fate of a new antitumour agent, 1-hexylcarbamoyl-5-fluoro[6-14C]uracil (¹⁴C-HCFU) was compared with that of 5-fluoro[6-14C]uracil (¹⁴C-FU) after oral administration to mice.

2. 1-(5-Hydroxyhexylcarbamoyl)-5-fluorouracil (5-hydroxy-HCFU) and 1-(5-oxohexylcarbamoyl)-5-fluorouracil (5-keto-HCFU) were found as major intermediate metabolites of ¹⁴C-HCFU and were produced by ω-1 oxidation.

3. FU was detected in plasma 180min after oral administration of ¹⁴C-HCFU, whereas unchanged FU disappeared within 60min after ¹⁴C-FU.

4. ¹⁴C-HCFU and resulting FU were retained in tissues for a long period after oral administration, while administered ¹⁴C-FU was rapidly degraded.