HPLC测定磷酸咯萘啶片中主药的含量

目的　建立测定磷酸咯萘啶含量的HPLC方法。方法　选用DiamonsilC1 8(2 50mm× 4 .6mm ,5μm)色谱柱 ,0 .0 8mol·L- 1 磷酸二氢钾 -乙腈 (85∶1 5)为流动相 (含 0 .1 %三乙胺 ,pH3 .0 ) ;检测波长 2 78nm ;流速为 1 .0ml·ml- 1 ;进样量 2 0 μl。 结果　磷酸咯萘啶在浓度 5 .0 8～ 40 6 .4μg·ml- 1 范围内线性关系良好 ;回归方程为A =- 1 2 91 8+1 8657C ,r =0 .9999;平均回收率为 99.8% ,RSD =0 .8% (n =5)。结论　本法简便、灵敏、准确     

HPLC法测定磷酸咯萘啶注射液的含量

目的：建立HPLC法测定磷酸咯萘啶注射液的方法。方法色谱柱为 Kromasil C18（150×4．6mm,5μm）;柱温30℃;以乙腈－0．3％三乙胺溶液（磷酸调pH至3．0）（15∶85）为流动相;流速为1．0mL? min －1;检测波长为277nm。采用外标法测定磷酸咯萘啶的含量。结果磷酸咯萘啶在20．0～160．0μg? mL －1范围内,线性关系良好（r＝1．0000,n＝5）,平均回收率为：100．3％,RSD＝0．2％。结论本方法操作简单,结果准确可靠,可以有效地用于控制磷酸咯萘啶注射液的药品质量。     

3H-咯萘啶在小鼠体内的分布和排泄

咯萘啶(PND)是一种疟原虫红内期裂殖体杀灭剂,与氯喹无交叉抗性,对恶性疟和间日疟均有显著疗效。用家兔的初步实验表明,PND原药在尿中的排泄量仅约为剂量的2%。我们已报告该药在体内分布较广。文献报道PND对鼠疟原虫红内期裂殖体有长效作用。为进一步了解该药的体内过程,对~3H-PND在小鼠体内的分布和排泄进行了研究。     

抗疟药咯萘啶在兔体内的药代动力学

本文报道抗疟药咯萘啶iv,im和ig给药后在兔体内的药代动力学。用NONLIN程序对血药—时间数据进行拟合。一次快速iv 6 mg/kg后的血药—时间过程符合线性三室开模型。药代动力学参数(±SD):t 1/2_β为59±10h;V_c2.418±0.287L/kg;V_d(ss),29±6 L/kg;总清除率Cl_T为0.442±0.131 L/kg·h。Im和ig给药后的动力学过程以线性二室开模型描述。im 6 mg/kg,吸收速率常数K_a为33.5±21.8 h^-1,t 1/2_β为52±8 h,吸收完全。Ig 30或60 mg/kg后的k_a为2.41±1.26 h^-1,t 1/2_β为55±5 h,吸收程度为34.6%。咯萘啶在血中呈不均一分布,im后1～96 h,球/浆浓度比为3～6。     

抗疟药咯萘啶有关化合物的合成及抗疟活性比较

合成了咯萘啶(Ⅰ)的有关化合物Ⅱ～Ⅴ,以探讨抗疟药咯萘啶化学结构中母环1位上氮杂原子及吡咯烷基Mannich碱侧链的存在,对该化合物抗疟作用的关系。经对有抗药性的疟原虫体内试验,合成的有关化合物Ⅱ～Ⅴ以及抗疟药氯喹和阿的平等的抗疟作用,均不如咯萘啶。提示上述氮杂原子及Mannich碱铡链的存在,对咯萘啶的抗疟作用,起着重要的和不可缺少的作用。     

抗疟新药咯萘啶及其类似物的合成

作者等根据一些抗疟药的构效关系,合成了一种新化合物2-甲氧基-7-氯-10-[3′,5′-双(四氢吡咯次甲基)一4′-羟基苯胺基]苯骈[b]1,5-萘啶(Ⅰ),代号7351,定名咯萘啶。它对红内期裂殖体的作用显著,毒性低。之后,又合成了它的类似物Ⅱ,这些类似物大多数对鼠疟Plasmodiumberghei的红内期均具有不同程度的作用,其中Ⅱ_1～6,9.10等的作用与Ⅰ相当。对子孢子诱发感染的鼠疟P.yoelii的作用以化合物Ⅰ、Ⅱ_{1,3,5,6,9,10,12,15}等为最强,优于伯喹对照组。值得注意的是,这类化合物既对血液转种的鼠疟P.berghei具有显著的作用,同时对子孢子感染的鼠疟P.yoelii也具有显著的作用。     

复方青蒿琥酯咯萘啶片中2种成分的红外光谱鉴别方法研究

目的:建立复方青蒿琥酯咯萘啶片中青蒿琥酯和咯萘啶的红外鉴别方法.方法:采用不同溶剂对复方青蒿琥酯咯萘啶片中的青蒿琥酯和咯萘啶成分进行提取纯化,分别测定两种组分的红外光谱图.结果:复方青蒿琥酯咯萘啶片经提取后,青蒿琥酯的红外光谱图与青蒿琥酯对照图谱和对照品图谱一致,咯萘啶的红外光谱图与同法处理的磷酸咯萘啶对照品光谱图一致...     

咯萘啶治疗恶性疟疾30例的疗效

本文报道用咯萘啶治疗恶性疟30例,其治疗剂量为2mg/kg,分别经2次或3次肌注;治疗剂量为3mg/kg,3次肌注。退热时间为26-36h,原虫转阴时间为41-57h。另用本品2mg/kg,肌注2次治疗间日疟12例,退热时间为18h,原虫转阴时间为37h。有脑型疟8例全部获得救治。有氯喹未治愈的2例患者改用本品后也能见效。本品对孕妇(2例)及婴儿(1例)均未出现不良反应。本研究证明咯萘啶抗疟效果好、副反应轻,值得在临床上继续试用。     

抗疟新药咯萘啶对大鼠胚胎毒性的观察

抗疟新药咯萘啶以1100 mg/kg/d×1,330 mg/kg/d×3,165 mg/kg/d×3和84 mg/kg/d×3四个剂量组分别于母鼠妊娠第7天(D₇)开始每日灌胃一次。另设空白对照组及阳性对照组(敌枯双5 mg/kg/d×7)。妊娠D₂₀杀鼠检查胚胎。结果表明:咯萘啶各组均引起胚胎早期吸收率升高,并随剂量递增而增加。该药还能延迟大鼠胚胎胸骨与枕骨的骨化。各试验组中未见胎鼠外形、内脏及其它骨骼畸形。可见咯萘啶对大鼠有较明显的胚胎毒性。     

一种新番荔枝内酯单体atemoyacin-B克服肿瘤多药抗药性(英文)

目的:探讨atemoyacin-B(Ate)克服肿瘤多药抗药性(MDR)作用及其机制,方法:Bullatacin(Bul)为阳性对照物,细胞毒测定以MTT法;P-gp功能测定以Fura 2-AM法;细胞内药物积累测定以荧光分光光度计法;细胞凋亡测定以流式细胞仪法,结果:Ate对MCF-7/Dox,MCF-7,KBvzoo和KB细胞的IC_(50)分别为122,120,1.34,1.27 nmol·L~(-1),Ate显著增加MDR细胞内Fura-2及多柔比星(Dox)的积累,但不增加相应敏感细胞的细胞内Fura-2及Dox的积累,Ate也能诱导MDR细胞凋亡.结论:MDR细胞对Ate同样敏感,不受抗药性影响,其机制与降低P-gp功能及增加细胞内药物积累有关。     

Modulation of multidrug resistance by three bisbenzyl-isoquinolines in comparison with verapamil

比较轮环藤碱（Ｃｙｃ）、海岛轮环藤碱（Ｉｎｓｒ）和海岛轮环藤酚碱（Ｉｎｓｎ）与维拉帕米（Ｖｅｒ）体外调节多药耐药性（ＭＤＲ）的作用．方法：细胞毒试验采用ＭＴＴ法，细胞内阿霉素（Ｄｏｘ）积累采用荧光分光光度法测定．结果：Ｃｙｃ，Ｉｎｓｒ，Ｉｎｓｎ和Ｖｅｒ在ＭＤＲ细胞系ＭＣＦ７／Ａｄｒ和ＫＢｖ２００能显著调节Ｄｏｘ和长春新碱的耐药性，且其作用呈剂量依赖性．Ｃｙｃ，Ｉｎｓｒ，Ｉｎｓｎ和Ｖｅｒ均能增加ＭＣＦ７／Ａｄｒ细胞内Ｄｏｘ的积累．Ｃｙｃ和Ｉｎｓｒ调节ＭＤＲ作用明显优于Ｖｅｒ，而Ｉｎｓｎ的作用类似于Ｖｅｒ．结论：Ｃｙｃ，Ｉｎｓｒ和Ｉｎｓｎ能通过增加ＭＤＲ细胞内Ｄｏｘ的积累而调节ＭＤＲ．     

米哚妥林对P糖蛋白介导的多药耐药的逆转作用

目的评价新型激酶抑制剂类抗癌药米哚妥林逆转P糖蛋白(P-gp)介导肿瘤细胞多药耐药的作用,并探讨其可能机制。方法米哚妥林0.5,1和5μmol·L-1分别加入P-gp高表达的K562/A02和K562细胞中培养72 h,MTS法测定细胞存活率以检测细胞毒性。米哚妥林0.125,0.25和0.5μmol·L-1在K562/A02和K562细胞中与无毒剂量的P-gp底物多柔比星、紫杉醇或长春新碱共培养72 h,MTS法测定细胞存活率以检测逆转耐药作用。米哚妥林0.5和10μmol·L-1与P-gp荧光底物罗丹明123在耐药和敏感细胞中共同孵育30 min后用流式细胞术分析底物积累的变化。米哚妥林0.5μmol·L-1与耐药和敏感细胞共同孵育72 h,Western印迹法检测耐药蛋白和信号分子的表达,定量PCR检测MDR1基因表达的变化。将米哚妥林与P-gp膜蛋白共孵育,化学发光法测定剩余ATP的量,检测米哚妥林对P-gp ATP酶活性的影响。结果米哚妥林对P-gp高表达的K562/A02和亲本敏感细胞K562的细胞毒性无明显的差异,其中米哚妥林0.5μmol·L-1时2种细胞的存活率均达80%以上。米哚妥林0.5μmol·L-1在细胞水平即可有效逆转K562/A02对多种底物的耐药,而对敏感细胞无显著作用;米哚妥林能显著抑制P-gp的外排作用,增加底物Rh-123在K562/A02细胞中的积累,且效果好于阴性对照维拉帕米0.5和10μmol·L-1;米哚妥林对P-gp基因和蛋白表达以及AKT和ERK的表达与磷酸化水平均无影响;米哚妥林对P-gp的ATP酶活性具有明显的抑制作用。结论米哚妥林可抑制P-gp介导的药物外排并逆转P-gp介导的肿瘤多药耐药。     

抗阿霉素的中国仓鼠卵巢上皮细胞系的抗药性

用细胞生物学和SDS-PAGE电泳方法,研究抗阿霉素(Dox)CHO细胞系(RCl)的特性。用间接免疫荧光法,未测到RCl超表达P-糖蛋白。RCl降低Dox的摄入,增加膜的流动性,Dox在CHO中主要分布在细胞核,在RCl中则分布在细胞质和细胞核。RCl中有一种分子质量为30—40 kDa的核蛋白的表达被抑制,而在敏感CHO中其表达正常。     

屈洛昔芬对K562耐阿霉素细胞株耐药性的逆转作用(英文)

目的:研究屈洛昔芬(DRO)对耐阿霉素(ADR)K562细胞株(K562/A02)多药耐药性(MDR)的逆转作用及逆转机制。方法:用DRO分别处理K562/A02和K562敏感株。MTT法观察DRO影响K562/A02对ADR化学敏感性的变化。DRO 10μmol/L处理K562/A02前后,通过RT-PCR和免疫细胞化学染色,分析MDR1、GSTπ基因表达的变化,采用流式细胞技术测定细胞内ADR浓度的变化。结果:DRO显著逆转K562/A02的MDR,在20、10和5μmol/L浓度时,对ADR的化学敏感性分别增加到14、13和4倍,逆转活性与维拉帕米相当。MDR1和GSTπ的mRNA和蛋白表达在DRO 10μmol/L处理后第2天开始下降,第5天明显降低。用20、10和5μmol/L浓度的DRO处理两株细胞,K562/A02细胞内ADR积累分别增加到2.9、2.3和1.5倍。但DRO不能明显增加K562细胞内的ADR的浓度。结论:DRO对K562/A02的MDR有较强的逆转活性,逆转强度与维拉帕米相当,其逆转机制有多种不同的途径。     

LY294,002, a specific inhibitor of PI3K/Akt kinase pathway, antagonizes P-glycoprotein-mediated multidrug resistance

The transmembrane transport pump P-glycoprotein (P-gp) causes the efflux of chemotherapeutic agents from cells and is an important system that secures multidrug resistance (MDR) of neoplastic cells. In the present study drug sensitive L1210 and multidrug resistant L1210/VCR mouse leukemic cell lines were used as an experimental model. We found that LY 294,002, a specific inhibitor of PI3K/Akt kinase pathway, reduced the degree of vincristine resistance in L1210/VCR cells significantly and in a concentration-dependent manner. This was accompanied by decrease in IC(50) value to vincristine from 3.195+/-0.447 to 1.898+/-0.676 micromol/l for 2 micromol/l, to 0.947+/-0.419 micromol/l for 4 micromol/l, and to 0.478+/-0.202 micromol/l for 8 micromol/l LY294,002. The IC(50) value of sensitive cells for vincristine was about 0.010 micromol/l. FACS analysis of the proportion of cells in apoptosis or necrosis by annexin-V apoptosis kit showed the following: (i) vincristine-induced apoptosis in resistant cell to a much lower extent than in sensitive cells; (ii) LY294,002 alone did not induce apoptosis or necrosis in both sensitive and resistant cells; (iii) LY294,002 applied together with vincristine significantly increased the number of apoptotic cells. Transport activity of P-gp in resistant cells was monitored using calcein/AM as substrate and was depressed by LY294,002 in a concentration dependent manner. Significant differences in calcein retention were not observed when cells were preincubated with LY294,002 at different times from 0.5 to 24h. Sensitive and resistant cells contain similar amounts of uncleaved (i.e., unactivated) caspase-3 but in latter cells the activation of caspase-3 by proteolytic cleavage was decreased. The reversal of vincristine resistance by LY294,002 was associated with marked activation of caspase-3. Western blot analysis revealed that the development of MDR phenotype in L1210/VCR cells was also associated with increased level of Bcl-2 protein. All the above findings point to the possible involvement of PI3K/Akt kinase pathway in modulation of P-gp mediated multidrug resistance in L1210/VCR mouse leukemic cell line. MDR reversal effect of LY294,002 is accompanied with this compound's influence on vincristine-induced apoptosis.     

Synthesis and biological evaluation of 131I‐labelled pyronaridine

¹³¹I‐Labelled pyronaridine, an antimalarial drug was synthesized and evaluated for its stability and biodistribution in rabbits. We observed that there was >99% yield of the labelled compound as determined by HPLC and radio‐TLC in our study. Dynamic imaging studies in rabbits showed that ¹³¹I‐labelled pyronaridine was not concentrated in the thyroid for up to 30 min as that of Na¹³¹I. Biodistribution studies in the blood, liver, heart, spleen, kidney and lung of rats revealed that the maximum percentage of the injected dose (% ID) and the percentage of the injected dose per gram organ weight (% ID/g) of ¹³¹I‐labelled pyronaridine was found in the blood and liver when compared to the other organs even after 6 days of the drug administration. Results suggest that ¹³¹I‐labelled pyronaridine can be effectively used as an imaging agent to study the metabolism and mode of a response of pyronaridine. Copyright © 2007 John Wiley & Sons, Ltd.     

R-dl-型维拉帕米调低KBv200肿瘤细胞对长春新碱和多柔比星的多药抗药性

目的:研究R-型维拉帕米对多药耐药的降低作用及其急性动物毒性,并与消旋维拉帕米的相应结果作比较。方法:细胞毒性的测定用MTT法;细胞内多柔比星积累的测定用萤光分光光度计法。急性毒性试验用BALB/c小鼠腹腔注射法。结果:R-型维拉帕米部分调低KBv200细胞对长春新碱和多柔比星的耐药性,其调低效应与作用浓度和作用时间有关。1.25μmol·L~(-1)的R-型维拉帕米与长春新碱对细胞作用24h,能够显著增加KBv200细胞对长春新碱的敏感性。在增敏和增加细胞内多柔比星累积方面,R-型维拉帕米与消旋维拉帕米效果一样,但R-型维拉帕米的急性动物毒性明显低于消旋维拉帕米。结论:R-型维拉帕米1.25μmol·L~(-1)提高耐药肿瘤细胞对长春新碱和多柔比星的敏感性,增加对长春新碱敏感性所需的药物作用时间可缩短至24h。     

Partial circumvention of P-glycoprotein-mediated multidrug resistance by doxorubicin-14-O-hemiadipate

Previously, we have reported partial circumvention ofP-glycoprotein (Pgp)-associated resistance to doxorubicin(Dox) in MCF7/R human breast carcinoma and P388/R murineleukemia cell lines by doxorubicin-14-O-hemiadipate (H-Dox)[Povarov L.S. et al. (1995) Russian J. Bioorganic Chemistry21: 797–803]. We felt that these changes were due toalterations in the cellular pharmacokinetics of the analog inmultidrug (MDR) resistant cells, as compared to that of Dox.To address this hypothesis, we performed comparative studiesof the accumulation, retention and intracellular localizationof H-Dox and Dox in Dox-sensitive murine leukemia cell lineP388/S and its Dox-selected, Pgp-positive drug resistantP388/R subline. These studies were performed in the presenceor absence of cyclosporin A (CsA), a competitive inhibitor ofPgp. Flow cytometric analysis revealed significant differencesin Dox and H-Dox accumulation in P388/R cells when compared toP388/S cells. In P388/R versus P388/S cells, there was a 38-fold decrease in Dox accumulation, but only a 5-fold decreasein H-Dox accumulation, indicating over a 7-fold increase in H-Dox buildup in resistant cells. CsA did not affect uptake orretention of either drug by sensitive cells. However,coincubation with CsA resulted in a 54-fold increase in Doxaccumulation and only a 5-fold increase in H-Dox uptake inP388/R cells, restoring anthracycline levels in P388/R to100% of that found in P388/S cells. Once internalized by theresistant cells, H-Dox was retained better than Dox regardlessof presence or absence of CsA. Confocal microscopic analysisrevealed the presence of H-Dox but no Dox in cellular nucleiof P388/R cells. Thus, increased activity of H-Dox towardP388/R cells was correlated with its enhanced ability to enterand be retained in these cells, and also with redistributionof H-Dox into the nuclei of the resistant cells as compared toDox. Overall, our findings support our initial hypothesis andprovide evidence that H-Dox, a 14-O-hemiadipate ofdoxorubicin, is affected by Pgp-mediated MDR to a lesserextent than parental Dox due to changes in the cellularpharmacokinetics of the analog.