5-氟尿嘧啶神经酰胺脂质体急性毒性的昼夜节律研究

目的研究5-氟尿嘧啶(5-FU)半乳糖神经酰胺(GalCer)脂质体急性毒性是否存在昼夜节律.方法分别于4、10、16、22时,采用静脉注射(iv),观察小鼠给予包封有5-FU的GalCer脂质体后所产生的毒性反应和死亡情况,计算半数致死量(LD50).结果iv 5-FU GalCer脂质体LD50分别为171.0、252.5、270.1和202.7 mg@kg-1;其95%可信区间分别为152.8～190.3、226.8～281.1、222.0～327.5和173.1～236.5 mg@kg-1.结论5-FU GalCer脂质体夜间的急性毒性大于白天,存在着明显的昼夜节律现象.     

齐多夫啶半乳糖神经酰胺脂质体的制备及理化性质研究

目的 制备齐多夫啶半乳糖神经酰胺脂质体并对其理化性质进行研究。方法 采用两次乳化法，以半乳糖神经酰胺为主要膜材制备齐多夫啶半乳糖神经酰胺脂质体并进行形态学观察、检测其包封率和稳定性。结果 透射电子显微镜下观察齐多夫啶半乳糖神经酰胺脂质体为粒径均匀的球形或近球形的小单层颗粒；粒径范围为60～270nm，D50为140nm；包封率达72．5％，但10d后快速下降。结论 齐多夫啶半乳糖神经酰胺脂质体制备方法可行，需进一步提高包封率和稳定性。     

姜黄素体内外逆转结肠癌的5-氟尿嘧啶耐药研究及机制探讨

目的 探究姜黄素在体内外对结肠癌5-氟尿嘧啶（5-fluorouracil,5-FU）耐药的逆转作用及其相关机制。方法 以SW480结肠癌细胞为亲代细胞,构建5-FU耐药细胞株SW480R,采用MTT法检测SW480R细胞的耐药指数以及不同浓度的姜黄素对SW480R细胞增殖能力的影响;流式细胞术检测姜黄素对SW480R细胞周期和凋亡的影响;Western blotting检测姜黄素对SW480R细胞上皮-间充质转化（epithelial-mesenchymal transition,EMT）相关蛋白以及Wnt信号通路相关蛋白的影响;采用裸鼠移植瘤模型检测肿瘤体积变化情况,计算姜黄素的体内抑瘤率,Western blotting检测肿瘤组织中相关蛋白变化。结果 SW480R的耐药指数为12.16,姜黄素能够剂量依赖性地抑制SW480R细胞的增殖,阻滞SW480R细胞周期于G₀/G₁期,以及诱导SW480R细胞凋亡;Western blotting结果表明姜黄素能够在体内外抑制EMT和Wnt信号通路的活性;体内试验表明姜黄素能够有效抑制裸鼠移植瘤的生长。结论 姜黄素能够在体内外逆转结肠癌的5-FU耐药,可能是通过调控Wnt信号通路从而抑制EMT的发生。     

5-氟尿嘧啶脂质体凝胶剂的制备和评价

目的 研制5-氟尿嘧啶(5-Fu)脂质体凝胶剂,并进行质量评价.方法 采用逆相蒸发-冻融法制备5-Fu脂质体,再用卡波普为基质制成凝胶剂;以离心法测定脂质体的包封率;以体外经皮渗透释药法,比较5-Fu脂质体凝胶剂及5-Fu普通凝胶剂中的经皮渗透作用.结果 5-Fu脂质体平均粒径为2.16±0.30μm,平均包封率为(56.17±2.52)%;体外透皮实验中,脂质体凝胶剂24h的药物浓度为158.6 mg·mL-1,明显高于普通凝胶剂91.2mg·mL-1.结论 载药脂质体凝胶剂可显著减少药物经皮吸收,可维持较长释药时间.     

齐多夫啶半乳糖神经酰胺脂质体在小鼠的药动学研究

目的　研究半乳糖神经酰胺脂质体包封的齐多夫啶(azidothymidine ,AZT)在小鼠体内的药动学。方法　以半乳糖神经酰胺 (galactosyceramide ,GalCer)为主要膜材制备齐多夫啶半乳糖神经酰胺脂质体 (AZT GalCer)并观察AZT Gal Cer及游离AZT在小鼠体内的药动学过程。结果　AZT GalCer组较游离AZT组血药浓度显著升高 ,特别是 30min后升高达数十倍 ;药物的Ke由 0 0 37h-1降至 0 0 2 1h-1,T1/2 由 19min延长至 35min ,AUC增加 2 .77倍。结论　齐多夫啶半乳糖神经酰胺脂质体可提高齐多夫啶的血药浓度 ,延缓药物消除速度。     

5-氟尿嘧啶温度敏感性脂质体制备方法的优化

用正交设计法L9（34，四因素三水平）筛选5－氟尿嘧啶温度敏感性脂质体的制备工艺。用100℃水浴加热导致制剂破坏的方法作为稳定性测定标准，对制备工艺进行筛选。最终选定5－氟尿嘧啶温度敏感性脂质体的制备方法为：取主药和各种辅料，用匀浆机7000r／min搅拌1h，再用超声波乳化器在超声强度（out－putl）、超声间隙（cycle）50％状态下，超声10min，同时用水浴冷却，控制脂质体溶液温度在30－40℃之间。     

冻融法制备5-氟尿嘧啶脂质体及其稳定性考察

采用反相蒸发法制备 5 氟尿嘧啶脂质体制剂 .结合冻融法 ,可使 5 氟尿嘧啶的包封率达(45 31± 0 2 0 85 ) % .离心加速实验和 ζ电位的测定进一步验证了冻融法可使脂质体稳定性提高 .     

5-氟尿嘧啶乙醇脂质体的改性及其透皮吸收研究

目的:研究胆固醇对5-氟尿嘧啶(5-FU)乙醇脂质体的改性及其体外透皮扩散的影响。方法:制备不同胆固醇含量的5-FU乙醇脂质体,并考察胆固醇含量对脂质体粒径、Zeta电位、分散指数、包封率、皮内药物滞留量等指标的影响。结果:加入胆固醇后粒径和Zeta电位变化不大,分散指数从0.584降至0.143,5-FU包封率从28.6%增至48.8%,皮内残留5-FU量从40%增至80%以上。结论:胆固醇不会改变乙醇脂质体的粒径大小及Zeta电位,但可提高其分散性和稳定性;加入适量的胆固醇可提高乙醇脂质体中5-FU的包封率及皮内药物滞留量。     

5-氟尿嘧啶乙醇脂质体和变形脂质体的制备及其体外透瘢痕研究

分别按照Touitou法和Cevc法制备5-氟尿嘧啶(1)乙醇脂质体和变形脂质体,并进行体外透瘢痕试验.建立了高效液相色谱法分析测定两者透过瘢痕组织的1累积含量及1在瘢痕组织中的滞留量.采用C18色谱柱,流动相为甲醇-水(5∶95),检测波长265 nm.结果表明:透瘢痕试验24 h后,1乙醇脂质体、1变形脂质体及1 PBS溶液透过瘢痕组织的1累积透过量(μg·ml-1·cm-2)和在瘢痕组织的1滞留量(μg/cm2)分别为14.12±0.1和10.74±1.17、12.35±1.21和17.48±0.82、3.08±0.57和3.21±0.19.可见纳米级乙醇脂质体和变形脂质体均能有效促进1进入并透过瘢痕组织.相比之下,乙醇脂质体具有更好的透瘢痕作用,而变形脂质体能使5-FU在瘢痕组织内具有较高的滞留量.     

乳腺癌耐受蛋白介导的5-氟尿嘧啶耐受及机制

目的筛选乳腺癌耐受蛋白(BCRP)介导的耐受药物,探讨BCRP介导抗肿瘤药物耐受的机制,优化以BCRP表达检测结果为评价指标,为肿瘤临床化疗方案提供有价值的资料。方法不同浓度的抗肿瘤药物分别处理BCRP呈不同程度表达的PA317/Teton/TREBCRP细胞,经MTT法筛选出BCRP介导的耐受药物。高效液相色谱仪检测耐受药物在细胞内的相对剂量。采用核DNA染料Hochest33258荧光染色技术和流式细胞术检测耐受药物诱导的细胞凋亡。结果随着细胞BCRP表达的增高,PA317/Teton/TREBCRP细胞对5氟尿嘧啶(5Fu)、甲氨蝶呤、多柔比星、吡柔比星、依托泊苷、米托蒽醌等药物的耐药指数增高(P<0.05),而对如紫杉醇、顺铂、长春碱、丝裂霉素、长春地辛等药物均敏感。细胞的BCRP表达量与胞内5Fu浓度具有负相关性(r=-0.885,P<0.05)。随着细胞BCRP表达的增高,经5Fu处理后细胞凋亡率随之降低(P<0.05),而Ko143能显著提高BCRP表达细胞的凋亡率(P<0.05)。结论BCRP能增强细胞对5Fu所致的凋亡耐受。     

Pharmacokinetics and tissue distribution of 5-fluorouracil encapsulated by galactosylceramide liposomes in mice

AIM: To study the pharmacokinetics and tissue distribution of 5-fluorouracil encapsulated by galactosylceramide liposomes (5-Fu-GCL) in mice. METHODS: The concentration of 5-fluorouracil (5-Fu) in serum was detected by high performance liquid chromatography after 5-Fu-GCL (80, 40, 20 mg/kg) and free 5-Fu (40 mg/kg) were injected intravenously into mice. The tissue distribution of 5-Fu-GCL (40 mg/kg) and free 5-Fu (40 mg/kg) was investigated, and concentration-time profile of the two preparations in the liver were studied. Data were analyzed by 3p97 program. RESULTS: Serum concentration-time curves of 5-Fu-GCL and free 5-Fu conformed to one compartment model of first order absorption. 5-Fu-GCL at 80, 40, and 20 mg/kg had T(1/2Ke) of 25.8+/-4.2, 27.3+/-4.4, and 28.2+/-5.6 min; C0 of 24.9+/-4.9, 17.7+/-3.6, and 11.5+/-2.7 mg/L; and AUC of 990.0+/-45.2,622.5+/-38.3, and 340.4+/-25.6 mg x min x L(-1), respectively. In contrast free 5-Fu at 40 kg/mg had T(1/2Ke) of 15.8+/-2.2 min, C0 of 35.8+/-6.2 mg/L, AUC of 639.0+/-35.9 mg x min x L(-1). The tissue distribution of 5-Fu-GCL in the liver and immune organs was significantly increased, while in heart and kidney it was remarkably decreased. The AUC of 5-Fu-GCL in the liver was 3 times higher than that of free 5-Fu. CONCLUSION: The pharmacokinetics and tissue distribution of 5-Fu-GCL appears to be linear-related and dose-dependent, and exhibits sustained-release and hepatic target characteristics.     

反相HPLC法测定人血浆中5-氟尿嘧啶的浓度

目的：建立RP-HPLC法测定人血浆中5-氟尿嘧啶的浓度。方法采用Agilent Eclipse XDB-C18色谱柱,流动相为0.02mol· L -1的磷酸二氢钾缓冲溶液（pH＝3.0）,流速为1.0mL· min -1,检测波长为264nm,柱温为25℃。结果5-氟尿嘧啶在2.0～30.0μg· mL -1度范围内呈良好的线性关系,检测限为4.0ng。5-氟尿嘧啶浓度为2.0、10.0、20.0μg· mL -1的日内RSD分别为3.6％、2.2％、1.1％,日间 RSD分别为5.2％、3.4％、1.8％,平均回收率分别为105.3％、97.2％、98.6％。结论该方法简便、快速、准确、重现性好,适用于5-氟尿嘧啶的药代动力学研究及临床血药浓度检测。     

共载多西他赛和维拉帕米脂质体逆转肿瘤耐药性的研究

研究共载多西他赛(docetaxel, DTX)和维拉帕米(verapamil, VRP)组成的脂质体(DTX-VRP liposome,DTX-VRP LP)对人乳腺癌化疗多药耐药性的逆转作用。采用薄膜分散法制备DTX-VRP LP;用激光粒度仪考察了其粒径大小和zeta电位;用超滤法测定了载药量和包封率;动态透析法考察脂质体在pH值7.4和6.8的磷酸盐缓冲液(PBS)中体外累积释放率;以DTX诱导的多药耐药人源乳腺癌(MCF-7/DTX)细胞株研究脂质体的体内外药效学。动物实验获得上海交通大学动物伦理委员会批准(批准号:2019-06^-172)。结果表明, DTX-VRP LP平均粒径约为140.9 nm, zeta电位约在-28.7 mV。DTX-VRP LP中DTX及VRP包封率和载药量分别是(81.7±3.9)%、(2.9±0.3)%和(59.6±0.6)%、(1.6±0.5)%;在pH 7.4和6.8 PBS中, 0～4 h内DTX-VRP LP组累积释放率分别约为40%和70%,相较于其他实验组, DTX-VRP LP组累积释放率稍慢些。体外药效学...     

Disposition of 5-fluorouracil during chronic hepatic arterial infusion to patients with carcinoma in the liver and effect on circulating platelets

5-Fluorouracil was administered by continuous hepatic intra-arterial infusion to eight patients with the diagnosis of cancer of the gastrointestinal tract and hepatic metastases. Its elimination characteristics were investigated to see if they correlated with therapeutic effect or reduced clinical toxicity when the drug was given by this route. Urinary excretion of drug and metabolites was similar to findings after intravenous bolus doses. Disposition changes could not be correlated with therapeutic effect or clinical toxicity. A dose-related biphasic effect of 5-fluorouracil was found on circulating platelets. Doses greater than 6 mg kg^?1 d^?1 decreased the number of circulating platelets, while doses less than that resulted in an increase in circulating platelets. Further studies are required to determine the mechanism of the effect of 5-fluorouracil on platelets.     

反相高效液相色谱法测定人血浆中氟尿嘧啶浓度

目的 研究测定人血浆中的氟尿嘧啶浓度的色谱方法。方法 色谱柱为C_(18)柱(大连依力特,250nm×4.6mm×5μm),柱温25℃,流动相为KH_2PO_4-H_3PO_4缓冲液(0.01M,pH2.95),波长269nm。结果 回归方程为Y=2.2368X+2.7756(r=0.9998),其线性范围为:5～2500μg/L,最低检测浓度为2μg/L(S/N=3)。结论 本方法简单、快速、准确,适用于临床氟尿嘧啶的浓度测定及治疗药物监测。     

中空纤维离心超滤结合高效液相色谱法测定5-氟尿嘧啶的血药浓度

目的:建立简单、快速的中空纤维离心超滤-高效液相色谱法(HFCF-UF-HPLC)测定人血浆中5-氟尿嘧啶血药浓度的方法.方法:将加入少量释放剂的血浆样本经中空纤维离心超滤预处理后,直接取20μL超滤液进行HPLC分析,采用Ve-nusil MP C18 色谱柱(250 mm×4.6 mm,5 μm),流动相:甲醇-0...     

Formulation and evaluation of cholesterol-rich nanoemulsion (LDE) for drug delivery potential of cholesteryl-maleoyl-5-fluorouracil

Abstract

It has been reported that cholesterol-rich nanoemulsions (LDE) can bind to low density lipoprotein (LDL) receptors which can concentrate anticancer drugs in the tissues via LDL receptor overexpression and reduced the adverse effects of the treatment. Therefore, in this study, LDE nanoemulsions of cholesteryl-maleoyl-5-fluorouracil (5-FU conjugate) were developed and evaluated in vitro. LDE nanoemulsions were prepared by high-energy emulsification technique. Developed formulations were characterized in terms of droplet size, polydispersity index, zeta potential, viscosity and refractive index. Optimized formulation (L5) was also evaluated for surface morphology using transmission electron microscopy (TEM). Developed formulations were subjected to in vitro drug release studies through dialysis membrane. The droplet size (50?nm), polydispersity index (0.109) and viscosity (32.16 cp) were found to be lowest for optimized formulation L5. The results of zeta potential indicated the stable formation of developed LDE nanoemulsions. TEM images of optimized formulation indicated non-spherical shape of droplets. About 97% of conjugate was found to be released from L5 after 24?h of study. Overall, these results indicated that developed LDE nanoemulsions could be successfully used for oral delivery of 5-FU conjugate.     

瘦素对5-氟尿嘧啶损伤结肠癌细胞的影响

目的探讨不同浓度的瘦素对5-氟尿嘧啶(5-FU)损伤结肠癌细胞的影响。方法同时用5-FU及不同浓度的瘦素进行体外干预结肠癌HT-29细胞株。MTT法检测5-FU50%细胞生长抑制率(IC50)的变化。流式细胞仪、原位末端转移酶标记法(TUNEL)进行周期及凋亡分析。以RT-PCR方法检测caspase-9,caspase-3mRNA的表达。结果瘦素抑制5-FU对HT-29的杀伤作用。抑制5-FU诱导的细胞周期阻滞及细胞凋亡。RT-PCR表明加入瘦素后caspase-9,caspase-3mRNA的表达均下降,且呈剂量依赖性。结论瘦素通过下调caspase-9,caspase-3的表达促进结肠癌细胞产生凋亡抵抗,抑制5-FU的损伤作用。     

Relationship between 5-fluorouracil exposure and outcome in patients receiving continuous venous infusion with or without concomitant radiotherapy

AIMS: Toxicity and response are correlated with plasma 5-fluorouracil (5-FU) concentration in patients treated with 5-FU at a dose of 1000 mg m(-2) day(-1). Head and neck cancer patients are treated with various therapeutic regimens, including chemotherapy with 5-FU at a dose of 600 mg m(-2) day(-1) with radiotherapy. We investigated the plasma concentration-effect relationship for this regimen, with the aim of developing recommendations for dose adjustment. METHODS: Patients received 5-FU at doses of 600 or 1000 mg m(-2) day(-1), as a continuous infusion over 4 or 5 days, with or without radiotherapy for the 600 mg m(-2) day(-1) regimen. The area under the curve (AUC) for 5-FU concentration was estimated, based on a single morning blood sample taken each day during treatment. AUC values were compared between patients with and without toxicity. This simplified method for AUC estimation was compared with the standard two-samples-per-day method in an independent group of 50 patients. RESULTS: Forty-six patients, corresponding to 115 courses, were included in this prospective study. Considerable interindividual variability in estimated AUC was observed for both doses. Grade 3-4 toxicity occurred in 10 and 21% of patients given doses of 600 and 1000 mg m(-2) day(-1), respectively. Ths study confirmed the relationship between plasma 5-FU concentration and toxicity previously reported for 1000 mg m(-2) day(-1), but found no such relationship for the 600 mg m(-2) day(-1) regimen with concomitant radiotherapy. CONCLUSIONS: Our results do not support the use of therapeutic drug monitoring to improve tolerance for the 600 mg m(-2) day(-1) regimen with concomitant radiotherapy. A simplified method is proposed for 5-FU monitoring for the 1000 mg m(-2) day(-1) regimen.