酒石酸长春瑞滨的细菌内毒素检查方法验证

目的参考USP、EP,探讨并建立酒石酸长春瑞滨的细菌内毒素标准与检查方法(凝胶法)。方法按《中国药典》(2010年版)二部附录ⅪE细菌内毒素检查方法进行。结果经验证,酒石酸长春瑞滨的细菌内毒素限值确定为L2.0IU/mg(以长春瑞滨计),通过稀释法可消除干扰,无干扰浓度为0.035mg/mL(以长春瑞滨计)。结论可按此标准采用本法对本品进行细菌内毒素检查。     

HPLC法测定肿瘤细胞中酒石酸长春瑞滨的浓度

目的:建立 HPLC 法测定肿瘤细胞中酒石酸长春瑞滨的浓度。方法:采用氰基柱(4.6 mm×250 mm,5μm),以乙腈-磷酸盐缓冲溶液(0.1 mol·L~(-1)磷酸二氢钾溶液,用磷酸调节 pH 至3.5)(25:75)为流动相,流速为1.0 mL·min~(-1),检测波长为268 nm,柱温25℃。结果:线性范围为0.0315～10.1μg·mL~(-1)。(r=0.9996);回收率(n=5)大于92.1%;日间、日内精密度试验的 RSD(n=5)均小于5.0%。结论:本试验建立的方法简便、快捷、专属性强,可用于临床研究酒石酸长春瑞滨的耐药机制,也可用于酒石酸长春瑞滨的药代动力学的研究。     

注射用重酒石酸长春瑞滨稳定性研究

目的考察注射用重酒石酸长春瑞滨的稳定性。方法采用高效液相色谱法测定其中重酒石酸长春瑞滨及有关物质的含量。以0.05mol/L磷酸缓冲液(取磷酸二氢钾6.8g,加800mL水溶解后,加三乙胺10mL,混匀,用磷酸调节pH=4.0,再加水至1000mL)-乙腈(68∶32)为流动相,检测波长为268nm。结果与放置0d时比较,低温(4±2)℃放置6个月后3批样品的各项指标均无明显变化。结论注射用重酒石酸长春瑞滨在低温条件下稳定性良好。     

蔗糖八硫酸酯三乙胺梯度法制备重酒石酸长春瑞滨脂质体

 目的：制备重酒石酸长春瑞滨脂质体,并对制备工艺进行考察。方法：以蔗糖八硫酸酯三乙胺（triethylammonium sucrose octasulfate,TEA8SOS）梯度法制备重酒石酸长春瑞滨脂质体,以阳离子交换树脂分离脂质体与游离药物;并以紫外分光光度法和激光粒度仪分别测定重酒石酸长春瑞滨脂质体的包封率和粒径。结果：重酒石酸长春瑞滨脂质体包封率约为95%,粒径为129.5 nm。结论：以蔗糖八硫酸酯三乙胺梯度法制备的重酒石酸长春瑞滨脂质体包封率较高,方法可行。     

离子载体A23187介导pH梯度法制备重酒石酸长春瑞滨长循环脂质体

目的:采用A23187制备重酒石酸长春瑞滨长循环脂质体,优化了处方工艺,并考察了含量、包封率、药脂比和体外释放等检测指标。方法:采用A23187介导的pH梯度法制备了重酒石酸长春瑞滨脂质体;用HPLC法检测了脂质体中重酒石酸长春瑞滨的含量和脂质(HSPC)的含量,考察了药脂比;采用阳离子交换树脂分离脂质体和游离药物,HPLC法检测包封率;以4 mmol.L-1NH4Cl-PBS(pH 7.4)为体外释放介质考察了脂质体的体外释放行为。结果:重酒石酸长春瑞滨脂质体包封率为96.1%,药脂比为1∶5(w/w);高药脂比有利于延长药物体外释放的时间。结论:采用A23187介导的pH梯度法制备重酒石酸长春瑞滨脂质体工艺可行、载药量大、包封率高;所建立体外释放的检测方法快速、准确。     

3-(S)-(1-氨甲酰基-1,1-二苯甲基)吡咯烷的合成

反式-4-羟基-L-脯氨酸经脱羧、N-Boc保护和氯代反应制得N-Boc-3-(S)-氯吡咯烷,在叔丁醇钾作用下与二苯乙腈缩合得N-Boc-3-(S)-(1-氰基-1,1-二苯甲基)毗咯烷,再经水解、脱保护、经L-(+)-酒石酸处理后碱化制得氢溴酸达非那新中间体3-(S)-(1-氨甲酰基-1,1-二苯基甲基)吡咯烷,总收率约14%.     

HPLC法测定酒石酸长春瑞滨脂质体注射液的含量

目的:建立HPLC法测定酒石酸长春瑞滨脂质体注射液中酒石酸长春瑞滨的含量。方法:采用Supecosil ABZ+Plus色谱柱(4.6 mm×150 mm,5μm),以0.05 mol·L-1磷酸二氢钠溶液(用磷酸调节pH至4.2)-0.2%癸烷磺酸钠的甲醇溶液(40∶60)为流动相,流速为1.0 mL·min-1,检测波长267 nm,柱温:35℃,进样量50μL。结果:在所选色谱条件下,主药与有关物质能较好分离,酒石酸长春瑞滨在0.02～0.06 mg·mL-1范围内,线性关系良好(r=0.9993);回收率范围为99.3%～100.1%(RSD=0.5%);检测限为1.7 ng;样品中含量测定符合质量要求。结论:该方法操作简便、灵敏、专属性强,准确度好,可用于酒石酸长春瑞滨脂质体注射液的质量控制。     

重酒石酸长春瑞滨联合5-氟尿嘧啶治疗胃癌的临床疗效观察

目的为了探讨重酒石酸长春瑞滨联合5-氟尿嘧啶治疗胃癌的临床治疗效果。方法总结在我院治疗的胃癌患者50例资料,按照患者自主选择治疗方法分为两组:选择重酒石酸长春瑞滨联合5-氟尿嘧啶进行治疗的26例资料为观察组,选择丝裂霉素联合5-氟尿嘧啶进行治疗的24例资料为对照组,两组患者治疗后按照文章疗效标准进行统计,最后统计学方法比较组间疗效差异性。结果观察组患者完全缓解2例(7.7%),部分缓解8例(30.8%),总有效率38.5%,与对照组疗效结果相当(P>0.05),而观察组治疗过程中不良反应情况明显优于对照组患者(P<0.05)。结论重酒石酸长春瑞滨联合5-氟尿嘧啶治疗胃癌具有满意的临床疗效,该治疗方案治疗过程中具有较轻的不良反应。     

不同厂家酒石酸长春瑞滨软胶囊质量对比研究

目的:对不同厂家酒石酸长春瑞滨软胶囊进行质量对比研究,评价不同厂家制剂的质量差异。方法:参考国家食品药品监督管理总局进口药品注册标准JX20130256,选择含量、有关物质、溶出度及不同贮藏温度下的稳定性等指标进行比较和评价。结果:不同厂家酒石酸长春瑞滨软胶囊制剂存在质量差异。结论:不同厂家制剂存在一定质量差异,建议有关厂家从制剂原辅料、制剂工艺等方面入手,不断提高制剂质量。     

注射用酒石酸长春瑞滨胶束及注射用长春瑞滨在比格犬体内的毒动学比较

目的 测定连续给予注射用酒石酸长春瑞滨胶束的毒动学参数，并与注射用酒石酸长春瑞滨进行比较。方法 选取16只比格犬随机分为4组，雌雄各半，分别连续iv给予注射用酒石酸长春瑞滨胶束低、中、高剂量（0.29、0.58、1.174 mg/kg）与注射用酒石酸长春瑞滨1.174 mg/kg，建立测定比格犬血浆中酒石酸长春瑞滨浓度的液相色谱-串联质谱（LC-MS/MS）法，测定血药浓度，采用DAS3.1.4药动学软件计算动力学参数。结果 比格犬iv给予注射用酒石酸长春瑞滨胶束低、中、高3个剂量，血药浓度、AUC_（0-t）、C_max随给药剂量的增加而增大；连续iv给药，低、中、高剂量组动物血药浓度、AUC_（0-t）、C_max在给药第1、29、71天时均变化不大，无明显蓄积倾向。而注射用长春瑞滨胶连续iv给药后随给药时间延长，动物血药浓度、AUC_（0-t）、C_max有上升趋势，AUC_（0-t）蓄积因子分别为2.08、1.80，C_max蓄积因子分别为2.58、2.32，均有蓄积倾向。结论 注射用酒石酸长春瑞滨胶束与普通注射用酒石酸长春瑞滨毒动学参数比较，无明显的蓄积倾向，可降低长期服药的毒性风险。     

The effect of tropolone on the formation of 3,4-dihydroxyphenylacetic acid and 4-hydroxy-3-methoxyphenylacetic acid in the brain of the mouse

1. The development of a very sensitive and specific fluorimetric assay for 3,4-dihydroxyphenylacetic acid has made it possible to measure how inhibitors of the enzyme catechol-O-methyl transferase affect the relative concentrations of this acid and its O-methylated derivative 4-hydroxy-3-methoxyphenylacetic acid (homovanillic acid) in the brains of mice treated with L-3,4-dihydroxyphenylalanine or probenecid.2. It was found that tropolone and tropolone-4-acetamide reduce the concentration of homovanillic acid in the brains of the treated mice to an extent dependent on the dose.3. The concentration of 3,4-dihydroxyphenylacetic acid in the brain was increased by the administration of tropolone or tropolone-4-acetamide but the dose and response were not simply related to one another.4. The results suggest that, in vivo, the formation of 3,4-dihydroxyphenylacetic acid is not always a simple alternative to the formation of homovanillic acid when the enzyme catechol-O-methyl transferase is inhibited.     

磺基水杨酸处理对高效液相色谱法检测人纤维蛋白原中蔗糖含量的影响

目的:研究磺基水杨酸处理样品对高效液相色谱法检测人纤维蛋白原中蔗糖含量的影响。方法:通过比较未经或经磺基水杨酸处理的蔗糖对照品溶液色谱图,确认磺基水杨酸处理对蔗糖水解的影响。蔗糖对照品溶液和人纤维蛋白原样品分别经磺基水杨酸处理2、3、4、5 h,研究处理时间对蔗糖水解的影响。用未经和经磺基水杨酸处理的蔗糖对照品溶液分别...     

Photocarcinogenesis study of glycolic acid and salicylic acid (CAS Nos. 79-14-1 and 69-72-7) in SKH-1 mice (simulated solar light and topical application study)

Acidic solutions have been used for decades to treat a variety of skin conditions. Many of these solutions consist of organic acids with a hydroxy group on a carbon adjacent to the carbonyl carbon and are referred to as alpha-hydroxy acids (AHA). Organic acids with hydroxy groups on the second carbon from the carbonyl carbon are referred to as beta-hydroxy acids (BHA). Both AHA and BHA are used to treat various skin conditions. One of the most widely used AHA is glycolic acid, while salicylic acid is a commonly used BHA. Chemical peels containing 20% to 70% glycolic acid have been used by dermatologists to treat ichthyosis, acne, xerosis, actinic keratosis, seborrheic keratoses, warts, and psoriasis. AHA have recently been used to treat photoaged skin and are now included in many commercially available cosmetic skin treatments. When used in a formulation for a chemical peel, topical treatment of skin with AHA and BHA can result in removal of the stratum corneum, alteration of the skin's histology, and increased cell proliferation in the basal layer of the epidermis. Since AHA and BHA are used to correct photoaged skin, and since exposure to sunlight of skin treated with AHA or BHA is likely, studies were designed to determine the effects of topical application of creams containing AHA (0%, 4%, or 10% glycolic acid, pH 3.5) or BHA (0%, 2%, or 4% salicylic acid, pH 4.0) on the photocarcinogenesis of simulated solar radiation using a filtered 6.5 kW xenon arc light source [simulated solar light (SSL)]. Male and female Crl:SKH-1 (hr-/hr-) hairless mice were exposed to glycolic acid or salicylic acid alone or in combination with SSL for 40 weeks, and the mice were followed for an additional 12 weeks. 1-YEAR STUDY IN MICE: Groups of 36 male and 36 female mice were exposed to 0.0, 0.3, 0.6, or 0.9 minimal erythema dose (MED) of SSL during the afternoon (1200 to 1600 hours) 5 days per week for 40 weeks. Groups of 18 male and 18 female mice were treated in the morning (0800 to 1100 hours) with 2 mg/cm2 control cream, 4% glycolic acid cream, 10% glycolic acid cream, 2% salicylic acid cream, or 4% salicylic acid cream on the dorsal skin, and in the afternoon (1200 to 1600 hours) with 0.3 MED of SSL 5 days per week for 40 weeks. Additional groups of 18 male and 18 female mice were treated in the morning (0800 to 1100 hours) with 2 mg/cm2 control cream, 4% glycolic acid cream, 10% glycolic acid cream, 2% salicylic acid cream, or 4% salicylic acid cream on the dorsal skin, and in the afternoon (1200 to 1600 hours) with 0.6 MED of SSL 5 days per week for 40 weeks. All mice were held an additional 12 weeks following the end of treatment. There were no effects of SSL exposure or topical treatment on the body weights of the mice. Increasing doses of SSL resulted in an SSL-dose trend in survival, with the greatest dose of SSL causing the earliest removal. This effect was present in both the untreated and control cream treated mice. The only consistent effect of glycolic acid on survival was a dose-dependent increase in survival of females at 0.3 MED SSL. Survival was increased in mice exposed to 0.6 MED of SSL and treated with 2% and 4% salicylic acid compared to mice treated with 0.6 MED and treated only with the vehicle. This effect was not observed in the mice treated with 0.0 and 0.3 MED of SSL and salicylic acid compared to the control groups. The mean or median time to first skin tumor of at least 1 mm decreased with increasing SSL exposure concentration in mice that were not treated with cream. Addition of the control cream resulted in a decrease in the time to tumor at 0.3 and 0.6 MED of SSL in male and female mice. The addition of glycolic acid (4% or 10%) did not affect the time to tumor in male or female mice at either SSL dose when compared to mice receiving the control cream. When compared to mice receiving control cream, the inclusion of 4% salicylic acid in the cream increased the time to tumor for male mice receiving 0.3 or 0.6 MED of SSL and female mice receiving 0.3 MED of SSL. The results indicate that inclusion of glycolic acid in the topical cream had no effect on the time required to induce tumors by SSL; however, inclusion of salicylic acid at 4% in the cream was photoprotective, increasing the time required to achieve median tumor incidence at a corresponding dose of SSL and control cream. The skin tumors induced by SSL in mice were squamous cell papilloma, carcinoma in situ, and squamous cell carcinoma. Except for papilloma in male mice, the tumors were induced in a dose-dependent manner by SSL in male and female mice. In male and female mice treated with control cream, the exposure to SSL caused significant increases in the incidences of carcinoma in situ, squamous cell carcinoma, and the combined incidence of carcinoma in situ and squamous cell carcinoma. When male or female mice were exposed to 0.3 or 0.6 MED SSL, the inclusion of 4% or 10% glycolic acid did not affect the induction of skin neoplasms over the incidence detected when the control cream was used, with the single exception of a glycolic acid dose-trend in squamous cell carcinoma incidence in male mice receiving 0.3 MED SSL. The inclusion of salicylic acid in the cream that was topically applied to female mice did not affect squamous cell papilloma formation at either SSL dose. The incidence of carcinoma in situ was decreased in male and female mice at 0.3 MED SSL when treated with 4% salicylic acid. A salicylic acid dose-trend was also observed in both sexes at 0.3 MED SSL. CONCLUSIONS: These experiments investigated the impact of topical application of a cosmetic formulation containing 4% or 10% glycolic acid (pH 3.5) or 2% or 4% salicylic acid (pH 4) on the photocarcinogenesis of filtered 6.5 kW xenon arc simulated solar light (SSL) in SKH-1 hairless mice. Taking into consideration the survival data, time to tumor data, and the pathology results, glycolic acid did not alter the photocarcinogenesis of SSL, and salicylic acid was photoprotective, reducing the carcinogenicity of 0.3 MED SSL.     

Oral eicosapentaenoic acid for acute colonic graft-versus-host disease after bone marrow transplantation

We investigated whether pretreatment with eicosapentaenoic acid, an inhibitor of leukotriene (LT) B4, could ameliorate acute colonic graft-versus-host disease (GVHD) after bone marrow transplantation (BMT). Seventeen patients undergoing unrelated BMT were divided into two groups, with eight patients receiving eicosapentaenoic acid and nine not receiving it. The grade of GVHD after transplantation was compared with that estimated from the pretransplantation LTB4 level. The levels of LTB4 and several cytokines were also monitored. The actual grade of GVHD was lower than that estimated from LTB4 levels in three of the eight patients from the treated group, and there was a significant difference between the treated and untreated groups (p < 0.05, chi 2 test). The levels of LTB4, tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) were all significantly lower in the treated group (p < 0.05, Student's t-test). These findings suggest that eicosapentaenoic acid may ameliorate acute colonic GVHD when administered from before BMT.     

卡立泊来德的合成工艺改进

以对异丙基苯甲酸为原料,经过氯磺化、还原、甲基化、胍解得到卡立泊来德,总收率为45·3%。改进后的工艺路线,降低了成本,简化了操作和反应条件。     

Lethality in mice of double-stranded ribonucleic acid from virus-like particles of Penicillium stoloniferum

The ld₅₀ in mice of intact virus-like particles (VLP) from Penicillium stoloniferum NRRL 5267 was greater than 240 mg/kg body weight, and their component double-stranded ribonucleic acid (dsRNA) was 34.0 mg/kg. Judged by decreases in ld₅₀ values, administration of sublethal amounts of actinomycin D or cycloheximide simultaneously with sublethal amounts of intact VLP or mycoviral dsRNA enhanced lethality in mice. A 2400-fold increase in the letahlity of intact VLP occurred in mice treated with actinomycin D and a 65-fold increase with cycloheximide and VLP. Increases in lethality of 850-fold with actinomycin D and 10-fold with cycloheximide were determined when mice were treated simultaneously with either antibiotic and mycoviral dsRNA. Similar lethality effects were observed in mice simultaneously treated with polyriboinosinic-polyribocytidylic acid and either of the antibiotics. Animals treated at zero-time with mycoviral dsRNA and subsequently treated at 1, 2, 4, 8, and 12 hr with actinomycin D generally exhibited a reduced toxic response after the 4-hr injection with actinomycin D. Animals treated at zero-time with actinomycin D and subsequently with mycoviral dsRNA also exhibited a reduced toxic response after the 4-hr injection with the nucleic acid.     

Effect of alpha-tocopherol on the microsomal lipid peroxidation induced by doxorubicin: influence of ascorbic acid

alpha-Tocopherol (40 mg/rat/day) was administered, orally, to doxorubicin treated rats (2 mg/kg, twice weekly, for 4 weeks) singly and also in combination with ascorbic acid (1 g/100 ml/day) in drinking water. The vitamin therapy was carried out for a period of 1 month. The microsomal lipid peroxide levels in liver and heart were found to be increased in doxorubicin treated rats. alpha-tocopherol and ascorbic acid treatment decreased the lipid peroxide level and also NADPH-dependent lipid peroxidation. A significant depletion of glutathione in liver and heart of doxorubicin treated animals was found to be ameliorated by vitamin therapy. Ascorbic acid was found to maintain the level of microsomal alpha-tocopherol. The activities of the detoxifying enzymes like catalase, superoxide dismutase and glutathione peroxidase were suppressed in doxorubicin treated rats and vitamins coadministration maintained the levels of these enzymes. Ascorbic acid was found to potentiate the antioxidant nature of alpha-tocopherol.     

Topical steroid treatment reduces arachidonic acid and leukotriene B4 in lesional skin of psoriasis.

Topical clobetasol propionate or vehicle ointment was applied daily for 3 days to psoriatic plaques on eight patients. Skin chamber exudates from untreated, steroid and vehicle treated lesions were assayed for arachidonic acid (AA), leukotriene B4 (LTB4), prostaglandin E2 (PGE2) and 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) before, and at 24 h and 72 h after treatment. Significant reductions in AA and LTB4 were observed at 72 h in steroid treated lesions. The reduction in 12-HETE levels observed after steroid treatment was not statistically significant. PGE2 levels in lesional psoriatic skin were unaltered. The reduction of AA, and LTB4 was associated with clinical improvement of psoriasis.     

Quinazoline antifolates inhibiting thymidylate synthase: 4-thio-substituted analogues

We report the synthesis of four new 4-thio-5,8-dideazafolic acid analogues and a 4-(methylthio) analogue structurally related to the thymidylate synthase (TS) inhibitor N10-propargyl-5,8-dideazafolic acid. Three N10-propargyl-4-thio-5,8-dideazafolic acid analogues had C2 amino, hydrogen, and methyl substituents. A 4-thio and a 4-(methylthio) compound each with hydrogen at C2 and ethyl at N10 were also synthesized. In general, the synthetic route involved thionation of the appropriate 4-oxoquinazoline; the sulfur thus introduced was then protected by methylation. Further protection with a pivaloyl group was required for the quinazoline bearing a 2-amino substituent. The protected quinazolines were treated with N-bromosuccinimide and the resulting 6-(bromomethyl) compounds were then coupled to the appropriate N-monoalkylated diethyl N-(4-aminobenzoyl)-L-glutamate in N,N-dimethylacetamide with calcium carbonate as base. The 4-thio-5,8-dideazafolic acids were obtained by removal of the methylthio group with sodium hydrosulfide, followed by deprotection of the carboxyl groups with cold dilute alkali. For the compound containing a pivaloyl protecting group, hot dilute alkali was used. To obtain the 5,8-dideazafolic acid containing a 4-(methylthio) substituent, the corresponding diester was treated with lithium hydroxide which selectively deprotected the carboxyl groups. The five compounds were tested as inhibitors of L1210 TS. It was found that replacement of the 4-oxygen of the quinazoline moiety by sulfur did not alter the TS inhibition. However, the introduction of a methylthio substituent at position 4 severely impaired TS inhibition. All 4-thio compounds were less cytotoxic to L1210 cells in culture than their 4-oxo counterparts.     

甘草甜素、甘草次酸与柴胡皂甙对防治大白鼠实验性肝硬化的作用

甘草甜素与甘草次酸能防治实验性肝硬化的发生,而柴胡皂甙则无此作用。为了进一步明确抑制肝纤维增生的机理,本文对其防治大白鼠实验性肝硬化的作用进行了进一步研究。结果表明,甘草甜素可明显阻止四氯化碳中毒大白鼠SGPT活力的升高,而其它二药则不明显。甘草甜素与柴胡皂甙能减少肝内甘油三酯的蓄积。病理组织学观察发现,经甘草甜素、甘草次酸治疗的大白鼠其肝损伤均较对照组为轻。组织化学观察显示,甘草次酸治疗的大白鼠肝糖原明显增多。甘草甜素与甘草次酸组的血清胎甲球蛋白检出率也高于对照组。实验结果提示,此三种药物无胶原溶解与重吸收的作用。