洛索洛芬钠渗透泵型控释片的研究 硕士论文(可编辑)

洛索洛芬钠渗透泵型控释片的研究 硕士(可编辑) 洛索洛芬钠渗透泵型控释片的研究 硕士论文 单位代码: 10369 学号: 200701102010届硕士研究生学位论文 洛索洛芬钠渗透泵型控释片的研究 STUDY ON LOXOPROFEN SODIUM OSMOTIC PUMP CONTROLLED RELAESETABLETS 学科专业药剂 学 研究方向药物新技术与新剂型 导 师王 成 永 教 授 硕 士 生杨 甜 2010年4月?合肥 目 录 中文摘 要 „„„„„„„„„„„„„„„„„„„„„„„„„„„ 1 英文摘 要 „„„„„„„„„„„„„„„„„„„„„„„„„„„ 3 英文缩略 词 „„„„„„„„„„„„„„„„„„„„„„„„„„ 5 前 言 „„„„„„„„„„„„„„„„„„„„„„„„„„„„„ 6 第一章 处方前研究及体外分析方法的建立 1 仪器与试 药 „„„„„„„„„„„„„„„„„„„„„„„„„ 13 2 方法与结 果 „„„„„„„„„„„„„„„„„„„„„„„„„ 13 2.1 处方前研 究 „„„„„„„„„„„„„„„„„„„„„„„„ 13 2.1.1 洛索洛芬钠的溶解 度„„„„„„„„„„„„„„„„„„„„ 13 2.1.2 洛索洛芬钠在不同释放介质中的稳定 性„„„„„„„„„„„„ 14 2.1.3 表观油水分配系数的测 定„„„„„„„„„„„„„„„„„„ 14 2.2 含量测定方法的建 立„„„„„„„„„„„„„„„„„„„„„ 15 2.2.1 检测波长的确 定„„„„„„„„„„„„„„„„„„„„„„ 15 2.2.2 色谱条件与系统适应 性„„„„„„„„„„„„„„„„„„„ 16 2.2.3 洛索洛芬钠对照品溶液的制 备„„„„„„„„„„„„„„„„ 17 2.2.4 曲线的制 备„„„„„„„„„„„„„„„„„„„„„„ 17 2.2.5 精密度试 验„„„„„„„„„„„„„„„„„„„„„„„„ 18 2.2.6 稳定性试 验„„„„„„„„„„„„„„„„„„„„„„„„ 19 2.2.7 重复性试 验„„„„„„„„„„„„„„„„„„„„„„„„ 19 2.2.8 回收率试 验„„„„„„„„„„„„„„„„„„„„„„„„ 20 2.2.9 洛索洛芬钠渗透泵片的含量测 定„„„„„„„„„„„„„„„ 20 2.3 释放度测定方法的建 立„„„„„„„„„„„„„„„„„„„„ 21 2.3.1 检测波长的确 定„„„„„„„„„„„„„„„„„„„„„„ 21 2.3.2 标准曲线的制 备„„„„„„„„„„„„„„„„„„„„„„ 22 2.3.3 精密度试 验„„„„„„„„„„„„„„„„„„„„„„„„ 22 2.3.4 释放介质中的稳定 性„„„„„„„„„„„„„„„„„„„„ 23 2.3.5 回收率试 验„„„„„„„„„„„„„„„„„„„„„„„„ 23 2.3.6 释放度测定方 法„„„„„„„„„„„„„„„„„„„„„„ 24 3 讨论 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 24 4 小结 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 25 第二章 洛索洛芬钠渗透泵型控释片制备工艺的研究 1 仪器与试 药 „„„„„„„„„„„„„„„„„„„„„„„„„ 26 2 方法与结 果 „„„„„„„„„„„„„„„„„„„„„„„„„ 26 2.1 洛索洛芬钠渗透泵型控释片的制备工艺及工艺流程图 „„„„„„ 26 2.1.1 制备工 艺„„„„„„„„„„„„„„„„„„„„„„„„„ 26 2.1.2 工艺流程 图„„„„„„„„„„„„„„„„„„„„„„„„ 27 2.2 洛索洛芬钠渗透泵型控释片的单因素考 察„„„„„„„„„„„„ 27 2.2.1 片芯处方的单因素考 察„„„„„„„„„„„„„„„„„„„ 27 2.2.2 包衣液处方的单因素考 察„„„„„„„„„„„„„„„„„„ 32 2.2.3 体外释放条件对药物释放的影 响„„„„„„„„„„„„„„„ 36 2.3 正交试验优化处 方„„„„„„„„„„„„„„„„„„„„„„ 38 2.4 处方的确 定„„„„„„„„„„„„„„„„„„„„„„„„„ 40 2.5 处方与工艺重复性考 察„„„„„„„„„„„„„„„„„„„„ 41 2.6 释药曲线的拟 合„„„„„„„„„„„„„„„„„„„„„„„ 41 2.7 释药机理的初步研 究„„„„„„„„„„„„„„„„„„„„„ 42 3 讨论 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 42 4 小结 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 44 第三章 洛索洛芬钠渗透泵型控释片的质量控制及稳定性研究 1 仪器与试 药 „„„„„„„„„„„„„„„„„„„„„„„„„ 45 2 方法与结 果 „„„„„„„„„„„„„„„„„„„„„„„„„ 45 2.1 外观性 状„„„„„„„„„„„„„„„„„„„„„„„„„„ 45 2.2 释放度检 查„„„„„„„„„„„„„„„„„„„„„„„„„ 46 2.2.1 方法学考 察„„„„„„„„„„„„„„„„„„„„„„„„ 46 2.2.2 释放曲线的绘 制„„„„„„„„„„„„„„„„„„„„„„ 46 2.2.3 释放度限度的确 定„„„„„„„„„„„„„„„„„„„„„ 46 2.3 样品的含量测定及其限 度„„„„„„„„„„„„„„„„„„„ 46 2.4 有关物质分析方法的建 立„„„„„„„„„„„„„„„„„„„ 47 2.4.1 色谱条 件„„„„„„„„„„„„„„„„„„„„„„„„„ 47 2.4.2 最低检测限和定量限的确 定„„„„„„„„„„„„„„„„„ 47 2.4.3 降解产物对主药测定的影 响„„„„„„„„„„„„„„„„„ 47 2.4.4 洛索洛芬钠渗透泵型控释片有关物质检 查„„„„„„„„„„„ 48 2.5 洛索洛芬钠渗透泵型控释片初步稳定性研 究„„„„„„„„„„„ 48 2.5.1 考察项目与检测方 法„„„„„„„„„„„„„„„„„„„„ 48 2.5.2 影响因素试 验„„„„„„„„„„„„„„„„„„„„„„„ 49 2.5.3 加速试 验„„„„„„„„„„„„„„„„„„„„„„„„„ 50 3 讨论 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 51 4 小结 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 52 第四章 洛索洛芬钠渗透泵型控释片在大鼠体内初步药动学研究1 仪器与试 药 „„„„„„„„„„„„„„„„„„„„„„„„„ 53 2 方法与结 果 „„„„„„„„„„„„„„„„„„„„„„„„„ 53 2.1 受试制剂与参比制 剂„„„„„„„„„„„„„„„„„„„„„ 53 2.2 实验动 物„„„„„„„„„„„„„„„„„„„„„„„„„„ 53 2.3 给药及血样采 集„„„„„„„„„„„„„„„„„„„„„ 53 2.4血浆样品的处理和测 定„„„„„„„„„„„„„„„„„„„„ 54 2.5 体内分析方法的建 立„„„„„„„„„„„„„„„„„„„„„ 54 2.5.1 色谱条 件„„„„„„„„„„„„„„„„„„„„„„„„„ 54 2.5.2 方法的专属 性„„„„„„„„„„„„„„„„„„„„„„„ 54 2.5.3 标准曲线的制 备„„„„„„„„„„„„„„„„„„„„„„ 55 2.5.4 精密度试 验„„„„„„„„„„„„„„„„„„„„„„„„ 56 2.5.5 样品回收率试 验„„„„„„„„„„„„„„„„„„„„„„ 56 2.5.6 血浆样品稳定性考 察„„„„„„„„„„„„„„„„„„„„ 57 2.6 血药浓度测定结 果„„„„„„„„„„„„„„„„„„„„„„ 58 2.7 相对生物利用 度„„„„„„„„„„„„„„„„„„„„„„„ 61 3 讨论 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 61 4 小结 „„„„„„„„„„„„„„„„„„„„„„„„„„„„ 62 全篇小 结 „„„„„„„„„„„„„„„„„„„„„„„„„„„ 63 参考文 献 „„„„„„„„„„„„„„„„„„„„„„„„„„„ 64 综 述 „„„„„„„„„„„„„„„„„„„„„„„„„„„„„ 68 个人简 历 „„„„„„„„„„„„„„„„„„„„„„„„„„„ 80 致 谢 „„„„„„„„„„„„„„„„„„„„„„„„„„„„„ 81 中文摘要 洛索洛芬钠(loxoprofen sodium)为苯丙酸类非甾体抗炎药,具有起效迅速,抗炎强效且均衡,胃肠道刺激小的特点,临床上广泛用于治疗慢性风湿性关节炎、类风湿性关节炎、骨性关节炎、变形性关节炎、腰疼等。目前,洛索洛芬钠已上市的品种有普通片、分散片、口腔速崩片、颗粒剂、胶囊剂、贴剂。国内尚未见自主研制的缓控释制剂上市。将洛索洛芬钠制成渗透泵型控释片能够在一定时间范围内以恒定的释药速度释放出一定量的治疗药物,可以减少给药次数和保持平稳的血药浓度,为病人提供一种相比较而言较为安全、方便的服药方式。 本文建立了紫外分光光度法用于渗透泵片释放度的测定,建立了高效液相色谱法用于渗透泵片含量测定和体内血药浓度的检测。以上分析方法准确可靠、方便快捷,满足分析要求。 本文在充分考察药物及各种辅料性质的基础上,以氯化钠、羟丙甲基纤维素K4M(HPMC K4M)、乳糖为片芯组成,以醋酸纤维素(CA)丙酮溶液、邻苯二甲酸二丁酯(DBP)和聚乙二醇400(PEG400)为包衣材料,制备了洛索洛芬钠渗透泵型控释片(loxoprofen sodium osmotic pump controlled release tablets)。从片芯处方、包衣液处方、体外释放条件等因素对洛索洛芬钠渗透泵型控释片的释药行为进行了单因素考察。在单因素考察及因素筛选的基础上,采用综合评分法,通过正交试验,优化处方。结果显示,优化的处方在12小时内呈零级释药特征,释药较完全,批间重现性良好。 为控制渗透泵型控释片的质量,根据洛索洛芬钠的理化性质和渗透型控释片的制剂特点,建立了本品的释放度、有关物质、含量的测定方法。结果显示方法准确、可行,能有效控制产品的质量,为制定该制剂的质量标准提供了一定的依据。参照药物稳定性试验指导原则的要求,以外观、释放度、含量、有关物质等项目作为考察指标,对洛索洛芬钠渗透泵型控释片进行影响因素试验(4500lx光照、60 ?高温及RH 92.5 %高湿)和加速试验。试验结果显示,各项考察指标在上述试验过程中均未发生明显变化,证明该制剂稳定性良好,本品的制备工艺研究和质量标准研究合理、方法可行。 本文以市售普通片作为对照,对洛索洛芬钠渗透泵型控释片进行了药动学研究。利用高效液相色谱法对体内血药浓度进行测定,两种制剂的峰浓度(C)分别为8.37、23.45μg/ml,达峰时间(Tpeak)分别为6.031、0.753h,药时曲线下面积(AUC)分别为62.197、54.936 μg/ml×h,其相对生物利用度为113.22%。结果表明,自制渗透泵片 具有出峰时间晚、峰值低、血药浓度平稳的特征。 关键词:洛索洛芬钠;渗透泵型控释片;质量标准;药代动力学 Abstract Loxoprofen sodium is a styrene-acrylic acid non-steroidal anti-inflammatory drugs with rapid onset, potent anti-inflammatory and balanced, gentle stimulation of the gastrointestinal tract, which is widely used in clinical treatment of chronic rheumatismarthritis, rheumatoid arthritis, osteoarthritis, deformation arthritis, back pain and so on. Currently, loxoprofen sodium varieties have been listed ordinary tablets, dispersible tablets, oral fast disintegrating tablets, granules, capsules, paste table.China has not yet independently developed controlled release formulations on the market.Made of the osmotic pump loxoprofen sodium controlled release tablet, it can release a constant drug release rate of a certain amount of therapeutic drugs within a certain time, which can reduce injecting times and maintain a stable blood concentration,To provide a comparison of patients is more safe and convenient way of taking medicine. It was established by UV spectrophotometry for the osmotic pump tablets release test, high performance liquid chromatography HPLC for the determination of osmotic pump tablet and the blood concentration of the test.The above analysis method is accurate, convenient, meet the analysis requirements. On the basis of investigating the characters of loxoprofen sodium and various ingredients, useing sodium chloride, HPMC K4M, lactose-chip core, with cellulose acetate CA acetone, dibutyl phthalateDBP and polyethylene glycol 400PEG400 for the coating material, prepared loxoprofen sodium osmotic pump controlled release tablets. For core with a slice prescription, clothing membrane prescription, in vitro release conditions as factors, loxoprofen sodium on osmotic pump tablets with drug release by the single factors studied. To single factor and factor-based screening, with integrated rate method, the orthogonal experimental design, optimization prescription.The results show that optimization of prescription drug release characteristics were zero-order in 12h, a more complete drug release, a good batch repeatability. According to the physico-chemical property of loxoprofen sodium and characteristic of osmotic pump tablets preparation, the drug release rate, related substances and content, which were the quality controls of loxoprofen sodium osmotic pump tablets. The result showed that the method was accurate and feasible, which provide some reference for quality control of loxoprofen sodium osmotic pump tablets. Referencing guidance principles of investigation on drug stability, the stability test of loxoprofen sodium osmotic pump tablets were carried4500 lx strong-light test, 60 ? high-temperature test, RH92.5 % high-humidity testby inspection of the character, drug release, related substance, content examination and so on. And then the acceleration test was carried on. The results indicated that loxoprofen sodium osmotic pump tablets were stable, nearly without any quality changes during the stability study. The preparation procedure and the quality control standard were reasonable and feasible. With the commercial loxoprofen sodium tablets as the reference, the pharmacokinetics was studied by self-prepare loxoprofen sodium osmotic pump tablets. HPLC was employed to detect the loxoprofen sodium concentration in blood plasma. Two preparations of peak concentration C were 8.37μg/ml and 23.45μg/ml, peak time Tpeak were 6.031 h and 0.753 h, area under the concentration-time curve AUC were 62.197μg/ml×h and 54.936μg/ml×h. The relative bioavailability of the two preparations was 113.22 %. The results show that osmotic pump tablets have property: evening peak time, peak low, stable plasma concentration. Key words: loxopeofen sodiumosmotic pump controlled tabletsquality standardpharmacokinetics 英文缩略词(Abbreviation) 缩略词 英文全名 中文释义 AUC area under concentration-time curve 药-时曲线下面 积 CA cellulose acetate 醋酸纤维素 d day 天 h hour 小时 HPLC high performance liquid chromatography 高效液相色 谱法 HPMC hypromellose 羟丙甲基纤维素 lx lux 勒克斯(光照度单位) min minute 分钟 MRT mean residence time 平均滞留时间 NSAIDs non-steroidal anti-inflammatory drugs 非甾体类 抗炎药 Papp apparent n-octyl alcohol/water partition 表观正辛 醇/水分配系数 PBS phosphate buffered saline 磷酸盐缓冲液 PEG polyethylene glycol 聚乙二醇 pH power of hydrogen 酸碱度 RH relative humidity 相对湿度 RSD relative standard deviation 相对标准偏差 前言 1 立依据与研究背景 1.1 研究背景 解热镇痛药是一类具有解热、镇痛作用,且大多数还具有显著的抗炎、抗风湿作用的药物,故称为解热镇痛抗炎药(antipyretic analgesic and anti-inflammatory drugs)。鉴于此本类药物的抗炎作用与具有甾体结构的糖皮质激素不同,1974年在意大利米兰召开的国际会议中,将本类药物归入非甾体抗炎药(non-steroidal anti-inflammatory drugs, NSAIDs)[1]。 自1899年德国拜尔公司研制出乙酰水杨酸(阿司匹林)acetylsalicylic acid, aspirin, ASA[2]问世以来的百年间,发现了许多不同类别的非甾体抗炎药,极大的丰富了临床治疗学,并广泛应用于临床。目前,非甾体抗炎药是全球处方药和非处方药用量最大的药物之一,在内、外、妇、儿科均广泛使用,在风湿病临床中也占有很重要的地位。 阿司匹林是非甾体抗炎药的原形药物,至今仍在使用,并且开发其新的用途。20世纪50年代出现的吡唑酮类药物,如保泰松,以及 20世纪60年代上市的吲哚乙酸类药物,如吲哚美辛(消炎痛),以其强大的抗炎、镇痛作用而广泛应用于临床。20世纪70年代至今,可谓非甾体抗炎药发展最快的年代,相继上市的丙酸类药物,如布洛芬和酮基布洛芬等;苯乙酸类药物,如双氯芬酸;萘乙酸类药物,如萘普生等。这些新型药物由于在化学结构上的变更,与阿司匹林或保泰松或吲哚美辛相比,疗效相当或更好,而不良反应的发生率和严重程度均明显下降。此外,为了提高疗效,同时减少其不良反应,出现了不同剂型的非甾体抗炎药,如戴芬(德国生产)是双氯灭酚的双释胶囊,布洛芬缓释剂型芬必得等[3]。 近年来,国内外已证实非甾体抗炎药主要的作用机制是抑制花生四烯酸arachidonicacid, AA代谢中的环氧化酶cyclooxygenase, COX和脂氧化酶lipoxygenase, LOX,阻碍前列腺素prostaglandin, PG、前列环素prostacyclin, PGI和白三烯leukotriene, LT的致炎增敏作用[4]。前列腺素和前列环素能使血管舒张并增强血管通透性,间接成为引起炎症的媒介,并且延长炎症时间,此外,前列腺素还可以使痛觉神经末梢增敏,产生疼痛。白三烯是过敏性慢反应物质SRS-A的主要成分,能增强血管通透性并促进血浆渗出。通过AA代谢所产生的大部分炎症介质均可为NSAIDs所阻断[5]。 除此之外,非甾体抗炎药的作用机制还包括:解除氧化磷酸化偶联;抑制溶酶体酶释放;从血浆蛋白里置换出内源性抗炎多肽;抑制补体活化;拮抗激酶活性及其产生;抑制氧自由基产生;抑制白细胞聚集和黏附[6]。 非甾体抗炎药的种类繁多,其“家族成员”还在不断增多,分类方法有很多种。根据对COX抑制特性将NSAIDs分为四类:第一类COX-1倾向性抑制剂,如小剂量肠溶性阿司匹林;第二类为非选择性COX抑制剂,如吲哚美辛、双氯芬酸等;第三类为选择性COX-2抑制剂,如美洛昔康等;第四类为COX-2特异性抑制剂,如罗非昔布、塞来昔布等[7]。还可以根据其对花生四烯酸的不同代谢途径来分类以及根据其化学结构来分类。 洛索洛芬钠(Loxoprofen Sodium)又称环氧洛芬钠、罗索普洛芬钠、氯索洛芬钠,由日本三共株式会社(Sankyo)研发,1986年7月在日本以商品名为“乐松”(Loxonin)上市。本品是第一个苯丙酸类前体型非甾体抗炎药,具有良好的镇痛、抗炎及解热作用,尤其是镇痛作用最强。其镇痛作用强度比吲哚美辛(消炎痛)强10倍,其抗炎和解热作用则与之相当。由于本品为前体药物,经胃肠道吸收后转变为活性代谢产物而发挥作用,故对胃肠道副作用少,使用更加安全。临床上广泛用于治疗慢性风湿性关节炎、类风湿性关节炎、骨性关节炎、变形性关节炎、腰疼等[1]。 1.2 立题依据 近年来,为增加药物的市场价值和竞争力,开发药物的新剂型成为重要的途径。在各种投放于市场的新型给药系统中,口服控释给药系统能够减小血药浓度的波动和给药次数,具有明显的给药方便性和较好的患者顺应性的优点。在各种口服控释给药系统中,渗透泵片独特的优点已成为研究开发的热点 [8-11]。一般的口服给药系统其 释药速率会受到释药环境的pH值、胃肠道蠕动的快慢等其它生理因素的影响,而渗透泵型控释片利用了渗透压原理释放药物,其释药行为对上述因素的变化不敏感,释药速度达到均匀恒定,体内外试验相关性良好[12],可同时控释多种药物[13]。因此,被公认是目前最理想的口服控释给药方法之一。 国内外研究报道表明,洛索洛芬钠显著的治疗效果和较小的副作用受到临床医生和患者的普遍认可,是我国“九五”及2010年医药科技发展规划推荐品种之一。在日本专利1997年到期后,福建汇天生物药业有限公司1998年开始对该品种进行研制开发,并于2002年7月获得国家新药证书和生产批件。目前,洛索洛芬钠已上市的剂型有普通片、分散片、口腔速崩片、颗粒剂、胶囊剂、贴剂。因此,积极开发推广本品的新剂型具有非常重要的现实意义。国内尚未见自主研制的缓控释制剂上市。将洛索洛芬钠制成渗透泵型控释片,能够在一定的时间范围内以恒定的释药速度释放出一定量的治疗药物,可以改善血药浓度的峰谷现象,保持平稳的血药浓度,同时减少给药次数,为病人提供一种相比较而言较为安全、方便的服药方式。所以本文选择洛索洛芬钠作为模型药物,制备渗透泵型控释片这一课题进行研究与开发,力求通过对控释制剂技术的研究,研制出设计合理,性质稳定,适合于生产,同时具有广大社会效益和经济效益的制剂新产品。 2 国内研究现状 2.1 洛索洛芬钠的基本理化性质 Chemical structures of Loxoprofen Sodium 化学名:Monosodium 2-{4-[2-oxocyclopentylmethyl]phenyl}propanoate 分子式:C15H17O3 Na分子量:268.28 本品为白色或浅黄色结晶或结晶性粉末,无气味,味微甜咸。在水、甲醇中易溶,在乙醇中溶解,在丙酮、氯仿、乙醚中不溶。 2.2 洛索洛芬钠的作用机制及药动学研究 洛索洛芬钠为苯丙酸类非甾体抗炎药,为非活性的前体药。吸收后在体内转化成活性代谢产物抑制环氧化酶,减少前列腺素生成,抑制中性粒细胞向炎症部位的趋向性及抑制趋向因子形成而发挥作用[14]。 本品口服后经胃肠道迅速吸收,转变为顺式和反式羟基代谢物,其药理活性是由反式-羟基代谢物产生[15,16]。健康成人单次口服洛索洛芬钠60mg,迅速吸收,在肝脏、肾脏、皮肤、其他细胞外间隙及四肢的炎性组织中浓度较高,在关节中浓度持续时间较长,而在脑组织和骨骼肌中浓度较低。服药后,原形药物的血药浓度达峰时间大约为30分钟,反式-羟基代谢物的血药浓度达峰时间大约为50分钟,给药1小时后的洛索洛芬钠原形及反式-羟基代谢物的血浆蛋白结合率分别为97.0%和92.8%。本品大部分以原形药物的葡萄糖醛酸的结合物或羟基化的葡萄糖醛酸结合物从尿液中排出。健康成人口服本品每次60mg,每日3次,连续5天,其吸收和排泄均与单次给药无明显差异,未见蓄积[17,18]。缪海均等[19]选取18例健康志愿者单剂量口服60 mg洛索洛芬钠片,采用高效液相层析法测定血浆中洛索洛钠药 物浓度,结果证明本品口服吸收快,消除半衰期短。 2.3 洛索洛芬钠的临床适应症 洛索洛芬钠具有起效迅速,抗炎强效且均衡,胃肠道刺激小的特点。临床上可广泛应用于各种急性或慢性关节炎,如风湿性关节炎、类风湿性关节炎、强直性脊柱炎、骨性关节炎和痛风性关节炎等;软组织风湿病,如腰疼、颈臂肩综合征、肩周炎、网球肘等;手术后、外伤后及拔牙后疼痛以及急性上呼吸道炎症的解热和镇痛[14]。黄烽等[20]选择52例活动性强直性脊柱炎患者,口服洛索洛芬钠,经过4周的治疗,总有效率为92.42%,其中对各主要疗效指标均有明显改善P0.05且耐受性较好。施冶青等[21]采用随机、开放、对照法,比较40例膝骨关节炎患者在治疗前后,膝关节活动痛、15 m行走时间、关节压痛度、日常活动能力等指标,结果显示均有显著性差异。 2.4 渗透泵片概况 渗透泵型控释制剂是以渗透压作为释药动力,以零级释放动力学为特征的一种释药系统。它不仅能够使血药浓度较长时间地维持在有效范围内,而且可减少用药次数与全身副作用,提高了药物的安全性和有效性。它具有如下优点[22-28]: (1)药物以零级速率释放,降低了峰谷现象,使药物因血药浓度波动而产生的毒副反应降低到最小,非常适合于治疗指数小的药物。 (2)相对于普通制剂药物恒速释放时间明显延长通常为12~24小时,因此可减少服药次数,提高患者顺应性,减轻刺激和不良反应,特别适用于半衰期短需频繁服用的药物。 (3)相对于其它缓控释制剂其释药速率受外界环境因素如胃肠道pH值、胃肠道蠕动、食物等影响小,因此个体差异小。 (4)体内外相关性好,可用体外释药参数预测体内释药情况。 关于渗透泵制剂的文献报到最早见于1955年,Rose等设计了一种利用渗透压差为释药动力的给药装置,主要用于埋植在动物体内,达到长时间定量释药的目的。但是由于该装置体积较大,使用不便。随着药剂学基本理论和制剂工艺学的发展,尤其是缓、控释制剂的不断开发应用,使人们对渗透泵制剂的研究逐步重视。20世纪70年代中期,F.Theeuwes利用醋酸纤维素作为包衣材料,无机盐或高分子聚合物作为渗透压促进剂,制备渗透泵型控释制剂,大大简化了渗透泵制剂的结构,使渗透泵制剂更加适合于工业化生产,从而使其从实验室走向市场,奠定了渗透泵制剂在缓、控释制剂中的特殊地位。 按照其结构特点,可将口服渗透泵片分为单层芯渗透泵片、双层芯渗透泵片和三层芯渗透泵片。单层芯渗透泵片的片芯一般包含药物、渗透压活性物质和助悬剂,外包一层控释半透膜,然后用激光或机械打孔的方法在片芯包衣膜上开一个释药小孔。口服后胃肠道的水分通过半透膜进入片芯,溶解片芯形成渗透压活性物质的饱和溶液及药物的饱和溶液或混悬液,使膜内具有很高的渗透压,在膜内外渗透压差的作用下水源源不断的进入片芯,在包衣膜内产生静压力,从而使药液以恒定速率压出释药孔[29],其流出量与渗透进入膜内的水量相等,直到包衣膜内外渗透压差为零。 对于一些难溶性药物,单层芯渗透泵片不能满足其理想释药 的目的,通常可制成双层芯渗透泵片,即在单层芯的基础上,在药物层上再加上一层推动层,推动层一般由高分子聚合物和渗透压活性物质组成,当渗透泵片处于水性环境时,水通过包衣膜进入片内,推动层高分子聚合物吸水膨胀,推动药物层经释药孔释药。甘勇等[30]以格列吡嗪为模型药物,采用了双层渗透泵技术,较好地实现了药物持续控制释放。 三层芯渗透泵片由助推层和两个药室组成,助推层置于中间。当助推层吸水膨胀后,推动药物经释药孔从两侧的药室释放出来[31]。由于双层芯渗透泵片在打孔前需要对药物层进行识别,这个过程速度慢而且辨识准确率也有待提高,而三层芯渗透泵片的片芯制备工艺复杂,相比之下,单层芯渗透泵片由于工艺简单而受到了广大药学工作者的青睐。 自1983年Alza公司的苯丙醇胺控释片(商品名:Acutrim)上市以来,已有20多个采用口服渗透控释技术的产品相继在国外上市,且多为1日给药1次,以初级单室渗透泵控释片和推拉式渗透泵为主。国内于20世纪80年代初首次发表了有关渗透泵控释制剂的研究论文, 1998年国内第一台用于科研和小批量生产的控释片剂激光打孔机研制成功[32]。近年来国内涌现出大量的相关研究报道,并且有数个产品上市或注册新药,如硝苯地平控释片(商品名:欣然)、硫酸沙丁胺醇控释片(商品名:惠百释)、格列吡嗪控释片(商品名:智唐)等。 3 课题研究内容与拟解决问题 3.1 课题研究内容 1 处方前研究及体外分析方法的建立 2 洛索洛芬钠渗透泵型控释片制备工艺的研究 3 洛索洛芬钠渗透泵型控释片的质量控制和初步稳定性的考察 4 洛索洛芬钠渗透泵型控释片在大鼠体内药物代谢动力学研究 3.2 课题拟解决问题 在现代药物缓控释理论的指导下,依照渗透泵片制剂原则,如何选用合适的辅料筛选出最佳的处方和制备工艺,制定相应的质量标准,是本课题研究的一个关键。药代动力学是新剂型发展的基础,剂型设计是否合理要由药代动力学的研究结果作出,对模型药物的剂型进行体内药动学研究,拟建立专属性强、灵敏度高、准确度好的体内测定方法,对该制剂作出体内评价以证明渗透泵型控释片的高生物利用度和控释效果。 因此,本文将分为体外试验和体内试验两部分:体外试验主要包括释放度试验测定方法的选择、含量测定方法的选择、制剂制备工艺的考察和优化,渗透泵型控释片体外释药机制的探讨以及制定相应的质量标准;体内试验主要进行了洛索洛芬钠渗透泵片的体内药物动力学研究,以期对洛索洛芬钠渗透泵片质量进行合理的评价并为临床合理用药提供科学的依据。 第一章 处方前研究及体外分析方法的建立 药物制剂的处方前研究(pharmaceutical preformulation studies)工作在处方设计和产品开发中是至关重要的,为选择最佳剂型、工艺和质量控制提供依据,使药物不但能保持其物理化学和生物 学稳定性,而且使制剂能获得较高的生物利用度和最佳疗效,关系到药物制剂的安全性、有效性、稳定性和可控性[33]。 本章建立了洛索洛芬钠的体外分析方法,并根据药物本身的性质、剂型和工艺的要求,考察了洛索洛芬钠的溶解性、稳定性、表观正辛醇/水分配系数(apparent n-octyl alcohol/water partition,Papp),为处方设计提供基础。 1 仪器与试药 日本岛津高效液相色谱仪(VP-ODS日本岛津色谱柱、LC-10AT输液泵、SPD-10A紫外检测器、CS-Light色谱工作站);TU-1901双束紫外可见分光光度计(北京普析通用仪器有限公司);AS10200A超声波清洗机(天津奥特赛恩斯仪器公司);AG285型电子天平(上海梅特勒?托利多公司);RCZ-SB药物溶出仪(天津大学无线电厂);ZD-88B恒温摇床(太仓市博莱特实验仪器厂)。 洛索洛芬钠对照品(中国药品生物制品检定所);洛索洛芬钠(江西金峰原料药有限公司);甲醇(色谱纯,天津市四有精细化学品有限公司);其它试剂均为分析纯。 2 方法与结果2.1 处方前研究 2.1.1 洛索洛芬钠的溶解性 按《中国药典》2005版二部凡例五(2)规定操作,取洛索洛芬钠适量,于25?2 ?一定容量的溶剂中,每隔5 min强力振摇30 s,30 min内观察溶解情况,如看不到溶质颗粒时,即认为已完全溶解。考察洛索洛芬钠在常用溶剂水、甲醇、乙醇、乙醚、氯仿、丙酮中的溶解 度情况,结果见Tab.1-1。结果表明,洛索洛芬钠在水、甲醇中极易溶 解,在乙醇中溶解,在乙醚、氯仿、丙酮中几乎不溶。 表1-1 洛索洛芬钠在不同溶剂中的溶解性 Tab.1-1 Solubility of loxoprofen sodium in different solvent Solvent Water Methanol Ethanol Aether Chloroform Acetone Solubility Easily soluble Easily soluble Soluble Insoluble Insoluble Insoluble 由于体外释放度的测定方法为37?,因此考察了37?条件下 洛索洛芬钠在不同释放介质中的溶解性,结果见Tab.1-2。结果表明, 洛索洛芬钠在蒸馏水、pH 6.8、pH 7.4磷酸盐缓冲溶液中极易溶解, 在0.1 mol/L HCl溶液中溶解。 表1-2 洛索洛芬钠在不同释放介质中的溶解性 Tab.1-2 Solubility of loxoprofen sodium in different release medium at 37? Solvent Water 0.1 mol/L HCl pH6.8 phosphate buffer pH7.4 phosphate buffer Solubility Easily soluble Soluble Easily soluble Easily soluble 2.1.2 洛索洛芬钠在不同释放介质中的稳定性 取洛索洛芬钠适量,加入水、0.1 mol/L HCl、pH 6.8磷酸盐 缓冲溶液,配制成浓度为20μg/ml的供试品溶液,于37?条件下放置 24 h,分别于0、1、2、4、6、8、12、24 h测定其吸收度,结果见Tab.1-3。 表1-3 洛索洛芬钠在不同释放介质中的稳定性 Tab.1-3 Stability of loxoprofen sodium in different release medium at 37? Medium A 0 1 2 4 6 8 12 24h Water 0.529 0.528 0.533 0.527 0.528 0.531 0.526 0.523 0.1 mol/L HCl 0.531 0.534 0.527 0.532 0.536 0.529 0.535 0.539 pH6.8 phosphate buffer 0.522 0.527 0.529 0.521 0.528 0.526 0.527 0.524 结果表明,洛索洛芬钠在水、0.1 mol/L HCl、pH 6.8磷酸盐 缓冲溶液中37?条件下24 h内稳定性良好,可以考虑作为释放介质。 2.1.3 表观油水分配系数的测定 本实验采用摇瓶法[34-35]测定洛索洛芬钠在不同pH值的氢 氧化钠-磷酸二氢钠缓冲溶液中的表观油水分配系数:精密称定干燥 至恒重的洛索洛芬钠对照品适量,用蒸馏水溶解,配制成0.2mg/ml溶 液,精密吸取该溶液2ml置于10ml容量瓶中,分别用不同pH值的氢氧 化钠-磷酸二氢钠缓冲溶液定容至刻度,作为供试液。精密量取5ml供试液以及等体积的水饱和正辛醇置于具塞三角瓶中,在37?恒温摇床中振荡24 h,静置15min,两相分层,取水相置于离心管中,3000r/min离心10min,用高效液相色谱法测定其浓度(Cw)。水相中原来的洛索洛芬钠浓度定为C0,萃取平衡后水相中的浓度定为Cw,则萃取平衡后油相中的浓度定为Co-Cw,表观油水分配系数为: Papp=Co-Cw/CwEqn.1-1 结果见Tab.1-4。结果表明,洛索洛芬钠的油水分配系数,随着pH值的增大,先增大后减小。 表1-4 洛索洛芬钠油水分配系数 Tab.1-4 Oil-water partition coefficient of loxoprofen sodium in different pH solution at 37? NO. pH1.0 pH2.3 pH4.0 pH6.8 pH7.4 Papp 85.05 198.31 22.13 0.72 0.29 2.2含量测定方法的建立 2.2.1 检测波长的确定 取一定浓度的洛索洛芬钠对照品溶液、以及按处方比例不含洛索洛芬钠的辅料在200~400 nm波长范围内进行扫描,结果见Fig1-1、Fig1-2。结果显示,洛索洛芬钠溶液在222 nm波长处有最大吸收,而辅料在此波长处无吸收,表明辅料无干扰,故选择222 nm作为检测波长。 图1-1 洛索洛芬钠对照品紫外扫描图 Fig.1-1 UV spectrum of loxoprofen sodium in solution 图1-2辅料紫外扫描图 Fig.1-2 UV spectrum of adjuvant in solution 2.2.2 色谱条件与系统适应性 色谱柱:日本岛津VP-ODS(150 mm×4.6 mm,5 μm); 流动相:甲醇-水-冰醋酸-三乙胺600?400?1?1; 检测波长:222 nm; 流速:1.00 ml/min;进样量:20μl;柱温:25?。 辅料的影响:取处方量的辅料,制成空白辅料溶液,注入色谱柱,记录色谱图。 在此条件下的洛索洛芬钠对照品、样品及阴性样品图谱见Fig1-3。洛索洛芬钠的保留时间为7. 9 min,阴性溶液对测定无干扰,理论塔板数以洛索洛芬钠计不低于2000。 A: Loxoprofen sodium control article B: Sample C: Blank excipient 图 1-3 洛索洛芬钠在不同样品中的色谱图(A:对照品,B:样 品,C:阴性对照) (1-洛索洛芬钠) Fig.1-3 HPLC chromatograms of loxoprofen sodium 1-Loxoprofen sodium 2.2.3 洛索洛芬钠对照品贮备液的制备 精密称取洛索洛芬钠对照品25.15 mg,置于25ml容量瓶中, 用水溶解并稀释至刻度,摇匀,得浓度为1.006mg/ml的对照品贮备 液。 2.2.4 标准曲线的制备 用移液枪精密吸取对照品贮备