【作者】 吴丽颖；

【导师】 曹德英；

【作者基本信息】 河北医科大学 ， 药剂学， 2008， 硕士

【摘要】 目的:间尼索地平(m-Nisoldipine,m-Nis)是河北医科大学药学院首次合成的国家一类新药,属于二氢吡啶类钙离子拮抗剂,为尼索地平的同分异构体。主要药理作用在于抑制钙离子进入可兴奋细胞,选择性地引起外周血管和冠脉血管的扩张,对心肌收缩力和心脏传导系统均无影响。主要用于治疗冠心病、心绞痛、高血压和慢性充血性心力衰竭及心源性休克。药理实验表明:间尼索地平的光稳定性增强,且具有长效、速效、强效的特点。根据该药物的处方前研究表明,其在酸性条件下稳定,将其制成胃内漂浮型缓释片,以增加药物吸收,稳定血药浓度,降低毒副反应和提高生物利用度,为临床研究及应用提供一种很有前景的缓释剂型。方法:在大量文献资料和预实验的基础上,以不同比例的阻滞剂硬脂酸和致孔剂聚乙二醇-6000(PEG-6000)与间尼索地平制成固体分散体(m-Nis-SD),并采用差示扫描量热法(DSC)验证固体分散体的形成。确定采用羟丙甲基纤维素(HPMC)为亲水凝胶骨架,加入助漂剂十八醇(C18H38O),发泡剂碳酸氢钠(NaHCO3)和(m-Nis-SD)以不同条件制备胃内漂浮型缓释片,考虑片剂漂浮性能的同时,进行影响药物释放速率的单因素考察,确定影响胃内漂浮型制剂中药物释放的主要因素为工艺因素(片剂硬度,干法和湿法,粘合剂)和上述几种处方因素。选取合适的制备工艺,以HPMC,聚乙二醇-6000,十八醇,碳酸氢钠进行四因素三水平的正交设计,综合评价片剂释放度和漂浮性能,通过极差分析,确定优化处方组成。间尼索地平胃内漂浮型缓释片的释放度测定方法,根据2005年版《中国药典》二部附录XD释放度测定法第一法要求,以250ml 0.1%十六烷基三甲基溴化铵的HCL溶液(9→1000)为释放介质,转速为100r·min-1,温度(37±0.5)℃,分别在2,4,6,8,10,12h取样5ml,滤过,随即补充等温等体积的空白介质,取续滤液,照分光光度法在237nm波长下测定吸光度A值,根据标准曲线方程计算药物浓度,求得间尼索地平胃内漂浮型缓释片的累积释放百分率,同法考察自制间尼索地平普通片的溶出度。以优化处方制备三批胃内漂浮型缓释片,进行含量测定和释放度试验,考察制备工艺稳定性,并将释放度数据分别用Higuchi模型,零级方程和一级方程进行拟合。将自制的间尼索地平胃内漂浮型缓释片(m-Nis-HBS)进行高温、高湿度和强光照射下的稳定性考察,以及密封自然放置条件下分别于制备后一个月、两个月、三个月和六个月时测定其漂浮性能和体外累积释放百分率,考察其稳定性。漂浮性能考察:于(37±0.5)℃的人工胃液中,采用搅拌桨转速为100r·min-1,模拟胃的蠕动,考察自制的间尼索地平胃内漂浮型缓释片的形态变化,起漂时间和持漂时间,并同法考察间尼索地平普通片的漂浮性能作对比。体内释药特性的研究以Beagle犬为实验动物,6只分两组(每组3只)研究m-Nis-HBS和对照普通速释片的体内药物动力学过程,给药后于不同时间点经Beagle犬股静脉取血样,经过预处理后,采用简便、快速、灵敏的高效液相色谱法,测定血浆中间尼索地平的浓度。测定结果用3p97药代动力学程序进行处理,模拟药-时曲线,并计算各项药代动力学参数。以Tmax、Cmax和AUC为指标,综合评价m-Nis-HBS和m-Nis普通片的相对生物利用度。同时以m-Nis-HBS不同时间的体外释放百分率Fr对其相应时间的体内吸收分数Fa进行回归,求回归直线方程和相关系数,考察m-Nis-HBS的体外释放与体内吸收的相关性。结果:通过DSC,证明了间尼索地平以非晶态分散于载体中,形成了固体分散体。单因素考察确定的制备工艺为:以75%乙醇为粘合剂,采用湿法制粒压片法制备m-Nis-HBS片,确定了处方各主要因素用量的水平。通过L9(34)正交实验设计得到优化处方为A2B2C1D1,HPMC35mg,硬脂酸-聚乙二醇-6000-间尼索地平(15:10:5)SD30mg,乳糖15mg,十八醇10mg,NaHCO310mg,根据极差大小可知,各因素对药物释放性能和漂浮性能的影响程度为C>A>B>D。制得的片剂表面光洁圆整,外观较好,硬度4～5kg。处方中HPMC遇水膨胀形成凝胶屏障,阻止内部片心进一步水化,可控制药物释放;硬脂酸和PEG-6000作为载体制成固体分散体,可使间尼索地平均匀分散,调节药物释放。十八醇为助漂剂,NaHCO3为发泡剂,均可显著提高片剂漂浮性能。根据优化处方制备了三批片剂,同一批6片取样时间点具均一性,三批片剂药物释放具有重现性,说明制备工艺稳定。将本品释放曲线分别用零级方程、Higuchi模型、一级方程进行拟合,回归方程分别为:说明释药特性更符合Higuchi模型,m-Nis-HBS片具有明显的缓释特征。释放度数据以Peppas方程进行拟合,释放机制为药物的扩散和骨架溶蚀协同作用。高温、高湿度、强光照射影响因素实验结果表明:m-Nis-HBS片对光不稳定,在生产和贮存过程中应严格避光保存;对温度和湿度较为稳定,虽然含量有所下降,但下降值均小于5%,片剂在RH75% 10d的吸湿增重小于5%,符合药典规定。以处方量80%、100%、120%的间尼索地平及相应辅料进行了回收率实验,回收率分别为:(99.85±0.23)%,(100.1±0.41)%,(99.91±0.24)%(n=3)。三批片剂含量测定结果为:(102.6±0.25)%,(101.1±0.56)%,(99.60±0.62)%(n=3),且含量均匀度合格。漂浮性能考察结果:于(37±0.5)℃的人工胃液中,在搅拌桨转速为100r·min-1情况下,自制的m-Nis-HBS均在3min内起漂且可持续漂浮12h以上。而m-Nis普通片3min内即全部崩解下沉。根据Beagle犬体内m-Nis-HBS片和对照普通速释片的体内药物动力学过程的研究结果,计算各项药动学参数。m- Nis-HBS和m-Nis普通片药动学参数分别为: T1/2Ka(h):1.509±0.1001,0.7684±0.1658 T1/2Ke(h):7.794±1.800,3.447±1.185 Tmax(h):5.854±0.5077,2.213±0.3225 Cmax(ng·ml-1):79.40±10.60,116.7±20.35 AUC(ng·ml-1)·h:1315±296.0,867.8±146.7 MRT(h):12.86±1.896,5.640±1.309二者体内过程均符合一级动力学一室模型。各项参数经配对t检验表明:消除速度常数Ke没有显著性差异(P>0.05);消除半衰期T1/2 (Ke)、吸收半衰期T1/2 (Ka)、峰浓度Cmax、达峰时间Tmax、药-时曲线下面积AUC均有显著性差异(P<0.05),说明m-Nis-HBS在Beagle犬体内具有明显的缓释制剂特征。以Tmax、Cmax和AUC为指标,综合评价m-Nis- HBS和m-Nis普通片的相对生物利用度。以m-Nis-HBS不同时间的体外释放百分率Fr对其体内的吸收分数Fa进行回归,得直线方程:Fa=0.8777Fr-7.486,r=0.9928,表明m-Nis- HBS的体内吸收与体外释放有显著的相关性,可以通过体外释放药时曲线预测体内吸收。结论:通过间尼索地平胃内漂浮型缓释片体外释放试验,表明m-Nis-HBS在人工胃液中可持续漂浮12h以上且药物释放符合Higuchi模型。Beagle犬体内药动学过程研究表明其在体内同样具有明显的缓释特征,可以为间尼索地平的临床研究和应用提供一种良好前景的缓释剂型。更多还原

【Abstract】 Objectives: m-Nisoldipine(m-Nis), a new isomer of nisoldipine(nis), is one of dihydropyridines calcium antagonists(DHP-CaA), being first developed by Department of Organic Chemistry, Hebei Medical University. Its main pharmacological action lies in suppresses the calcium ion to enter may the excitatory cells, selectively causes the circumference blood vessel and the crown arteries blood vessel’s expansion, does not have the influence to the cardiac muscle shrinkage force and the heart conduction system. Mainly uses in treating coronary disease, the angina pectoris, hypertension and the chronic congested heart failure and the heart source shock. Pharmacology experiment showed: m-Nis was more stable than nis to light and it was ultralente, prompt and powerful. According to m-Nis’s pre-preparation research, it is stable in acid medium. In order to increase m-Nis’s in vivo absorption, steady the drug concentration in blood, reduce its adverse effects, and enhance its bioavailability, the intragastric floating sustained-release tablet was prepared with HPMC hydrophilic matrix. It will provide a potential sustained-release dosege form for m-Nis’s clinical research and application. Methods: On the basis of scientific materials and the pre-preparation investigation, we select HPMC as the hydrophilic matrix, add PEG-6000 as the porogeneous compound, stearic acid and stearylalcohol as the retarder, and NaHCO3 as the gas-generating agent in the formulation. Considering the release properties and floating ability of the tablet, We carried through monofactorial investigations. After choosing suitable preparation technics, we use L9(34) orthogonal design to select the optimum formulation in terms of the floating capabilities and in vitro accumulative release percentage.Paddle agitation was used in the experiment of in vitro release test. The release test is carried out in simulated gastric fluid HCL(9→1000) with rotation rate 100r·min-1 at temperature (37±0.5)℃. Samples were withdrawn periodically at 2, 4, 6, 8, 10, 12h from the release medium, replensihed with the same volume of fresh medium each time, and then analyzed and determined for their drug contents at 237nm by ultraviolet spectrophotometer. The conventional m-Nis tablet was studied by the same means. Three batches of m-Nis-HBS were prepared and studied through release experiment to investigate the stability of formulation and technics. The release data were analyzed with three models: Higuchi equation, zero order kinetics, and first order kinetics. We studied the stability of m-Nis-HBS under following circumstances: high humidities, high temperature, strong light and natural store condition. To elucidate the mechanism of drug release we used Peppas equation to analyze the release data. Its floating properties was invertigated in (37±0.5)℃0.1molL-1HCL under 100r·min-1 paddle agitation.In vivo study: The intragastric residence experiment was carried in Beagle dogs. The release properties in vivo were investigated as following: 6 Beagle dogs were divided into two groups. One was given m-Nis-HBS, and the other group was given the conventional tablet. Plasma samples were obtained at different time after the administration. The plasma concentration of m-Nis was determined by High performance Liquid Chromatograph(HPLC) with Ultraviolet detector.Results: Through the L9(34) orthogonal design, the optimum formulation was founded: A2B2C1D1, that is: HPMC(A) 35mg stearic acid-PEG-6000(B)-m-Nis(15:10:5)SD 30mg stearyl alcohol(C) 10mg NaHCO3(D) 10mg Lactose 15mgThe factors influence the tablet’s release and floating properties as following turn: C>A>B>D. Three batches HBS tablets were prepared according to the optimum formulation. The release data were analyzed with three models: Higuchi equation, zero and first order kinetics, the regression equations: Q=7.6486t+9.16, r=0.9909 Q=37.857t1/2-33.952, r=0.9966 lnQ=0.1456t+3.0022, r=0.9536 The results showed that m-Nis-HBS conformed to Higuchi equation better than the other two models. The drug release mechanism was non-Fick’s diffusion, that was concurrence of the matrix’s dissolution and the drug’s diffusion. The HBS tablet was sensitive to high humidity, high temperature and strong light. The recoveries of equivalent 80%, 100%, 120% of the optimum formula were (99.85±0.23)%, (100.1±0.41)%, (99.91±0.24)% respectively. The contents of three batches of m-Nis-HBS tablet were (102.6±0.25)%, (101.1±0.56)%, (99.60±0.62)% (n=3). The content uniformity of the three batches optimum formula were within the stated range(n=10). Buoyancy test: the HBS tablet can set on floating within three minutes and maintain floating for more than twelve hours in each condition above. While the conventional tablet dissolved in less than three minutes and subsidised.The result of the pharmacokinetics in Beagle dogs shows that m-Nis-HBS and the conventional tablet both conformed to one compartment model. Pharmacokinetics parameters of m-Nis-HBS and m-Nis conventional tablet were respectively as following: T1/2Ka(h):1.509±0.1001,0.7684±0.1658 T1/2Ke(h):7.794±1.800,3.447±1.185 Tmax(h):5.854±0.5077,2.213±0.3225 Cmax(ng·ml-1):79.40±10.60,116.7±20.35 AUC(ng·ml-1)·h:1315±296.0,867.8±146.7 MRT(h):12.86±1.896,5.640±1.309 The result of t-test showed significant difference between the parameters T1/2(Ka), Tmax, Cmax, AUC and MRT of the two groups(P<0.05). The relative bioavailability of m-Nis-HBS and m-Nis conventional tablet is 156.2%. The vitro release percentage and the vivo absorption percentage showed significant correlation, the regression equation was: Fa=0.8777Fr-7.486, r=0.9928.Conclusions: Through the release experiment in vitro and pharmacokinetics study in Beagle dogs, we know that the m-Nis-HBS tablet showed obvious sustained-release properties in both vitro and vivo, it will provide a potentially promising preparation for the clinical research and applications of m-Nisoldipine.更多还原