辛苯聚醇阴道用壳聚糖温敏缓释凝胶的研制

发布时间：2018-08-15 13:43

【摘要】：辛苯聚醇(Octoxynol-9,O-9)是应用广泛的壬苯醇醚(Nonoxynol-9,N-9)的类似物,也是一种非离子型表面活性剂类阴道用杀精剂,但其毒性较N-9小[1-2]。而现今的外用避孕制剂多存在易于泄漏,难以铺展覆盖阴道表面,在阴道内滞留时间短,生物利用度低的问题。温敏凝胶是在位凝胶的一种,可响应温度变化发生相变而具有独特的溶液-凝胶转变性质,从而有着给药方便、黏附力强、长时间滞留的特点,因此尤其适用于黏膜给药[3-4]。壳聚糖(Chitosan,CS)也是温敏凝胶体系中一种常见的材料,有着良好的生物降解性、生物相容性、无毒与抑菌性,是一种优良的生物粘附性材料,有延缓药物释放的作用[5-6]。因此本研究将安全性更高的O-9与优良的生物粘附材料CS结合,制备具有温度敏感性的新型阴道用制剂,可使制剂在阴道表面广泛覆盖,延长药物在阴道中的滞留时间,得到更安全、有效、使用方便的新型外用避孕制剂。本研究选择药用辅料泊洛沙姆(Poloxamer)与CS结合作为温敏凝胶基质,采用冷溶法制备辛苯聚醇阴道用温敏性凝胶[(O-9)-VTG],并用星点设计-效应面法优化处方,筛选出最优处方的基质配比为P407:P188:甘油:CS=16.3:5.7:5:0.6,按处方制备的(O-9)-VTG的胶凝温度为33℃,胶凝时间约1.6min。采用高效液相色谱法建立了(O-9)-VTG质量控制方法,并对3批样品进行了质量评价,含量分别为标示量的99.83%、100.69%、99.31%(n=3)。为探讨(O-9)-VTG的体外释放行为,采用无膜溶出法进行体外释放度的研究。结果表明,(O-9)-VTG释药模型拟合遵循零级方程。通过影响因素试验、加速实验及长期稳定性试验,以外观性状、pH、胶凝温度、O-9含量为指标对制剂进行初步稳定性考察,结果表明(O-9)-VTG具有良好的稳定性。采用试管倒置法可测得胶凝完全时的胶凝温度,但无法对凝胶的相变行为及凝胶强度等进行表征,因此采用动态流变学实验测定温敏凝胶在相变过程中的流变参数,可测得(O-9)-VTG的胶凝温度为32.6℃,与试管倒置法测得结果基本一致,其在室温下(25℃)具有良好的流动性,粘度小,为牛顿流体,易于注射入阴道内;置于阴道后可迅速铺展,在一定时间内胶凝后,凝胶粘度增大,具有假塑性流体特征。另外模拟了阴道液的稀释作用对(O-9)-VTG胶凝行为的影响,证实了(O-9)-VTG在体温条件下仍可形成凝胶。通过分别阴道给予雌性大鼠自制辛苯聚醇普通凝胶[(O-9)-gels]及(O-9)-VTG,收集含药阴道冲洗液,建立阴道冲洗液中药物含量测定方法,并采用DAS软件对阴道冲洗液中药物量进行处理得到(O-9)-gels和(O-9)-VTG的阴道内滞留时间,结果与(O-9)-gels相比,制备的(O-9)-VTG具有明显的阴道内粘附滞留性能。为评价制剂的有效性,以市售壬苯醇醚凝胶(N-9 Jelly)为对照,采用改良的Sander-cramer方法观察(O-9)-VTG的最低有效杀精浓度(MEC)对大鼠精子的作用,结果表明,(O-9)-VTG的MEC为0.50mg·ml-1,与N-9 Jelly的效果相当。大鼠阴道内给药后,使雌雄鼠合笼,观察避孕效果,得到(O-9)-VTG与N-9 Jelly均有较高的避孕率的结论,与自然对照组及空白凝胶组之间的差异具有显著性,表明(O-9)-VTG阴道用药具有较好的抗生育作用。阴道粘膜刺激性试验及阴道急性毒性试验以SD雌性大鼠为对象,通过肉眼和阴道病理切片观察,采用Eckstein刺激性评价标准评价刺激性,结果显示(O-9)-VTG对大鼠阴道无明显的刺激作用,局部急性毒性实验表明经阴道用药也无毒性。综上所述,本研究制备的(O-9)-VTG胶凝温度适宜,温度敏感性好,制剂质量稳定,具有一定的缓释作用,体内滞留时间长,组织相容性好,制备得到更安全、有效、使用方便的新型外用避孕药,也为辛苯聚醇阴道局部用药制剂的进一步开发研究提供依据和基础。
[Abstract]:Octoxynol-9 (O-9) is a widely used analogue of nonoxynol-9 (N-9) and a nonionic surfactant vaginal spermicide, but its toxicity is less than that of N-9 [1-2]. Temperature-sensitive gel is a kind of in-situ gel, which can respond to temperature change and has a unique solution-gel transition properties, so it has the characteristics of convenient administration, strong adhesion and long retention, so it is especially suitable for mucosal administration [3-4]. Chitosan (CS) is also a common kind of temperature-sensitive gel system. Material, with good biodegradability, biocompatibility, non-toxicity and bacteriostasis, is an excellent bioadhesive material, has the effect of delaying drug release [5-6].Therefore, this study will be safer O-9 and excellent bioadhesive material CS combined to prepare a new temperature-sensitive vaginal preparation, can make the preparation in the negative. A new type of external contraceptives with more safety, effectiveness and convenience was obtained by extending the drug retention time in vagina. In this study, the thermosensitive gel [(O-9) -VTG] for octylene polyol vagina was prepared by cold-dissolving method with the combination of Poloxamer and CS as the thermosensitive gel matrix. The optimal matrix ratio was P407:P188:glycerol:CS=16.3:5.7:5:0.6. The gelling temperature and gelling time of (O-9) -VTG prepared according to the prescription were 33 C and 1.6 min. The quality control method of (O-9) -VTG was established by high performance liquid chromatography, and the quality of three batches of samples was evaluated. In order to investigate the release behavior of (O-9) - VTG in vitro, the release rate of (O-9) - VTG in vitro was studied by membrane-free dissolution method. The results showed that (O-9) - VTG release model followed zero-order equation. The results showed that (O-9) - VTG had good stability. The gelation temperature of the gel could be measured by inverted tube method, but the phase transition behavior and gel strength could not be characterized. Therefore, the rheological parameters of the thermosensitive gel during phase transition could be measured by dynamic rheological experiment. The gelling temperature of 9-VTG is 32.6, which is basically the same as that measured by tube inversion method. It has good fluidity at room temperature (25) and low viscosity. It is a Newtonian fluid and is easy to be injected into vagina. It can be spread quickly after being put into vagina. After gelling for a certain period of time, the gel viscosity increases and has the characteristics of pseudoplastic fluid. The effect of dilution of vaginal fluid on the gelation behavior of (O-9) - VTG proved that (O-9) - VTG could still form gel at body temperature. The normal gel [(O-9) - gels] and (O-9) - VTG were administered to female rats vaginally respectively, and the vaginal flushing fluid containing drugs was collected to establish a method for the determination of drug content in vaginal flushing fluid. The intravaginal retention time of (O-9) - gels and (O-9) - VTG in vaginal flushing solution was compared with that of (O-9) - gels. To evaluate the effectiveness of the preparation, the modified Sander-Cramer method was used to observe the intravaginal retention of (O-9) - VTG. The effect of the lowest effective spermicidal concentration (MEC) of (O-9) - VTG on rat spermatozoa was studied. The results showed that the MEC of (O-9) - VTG was 0.50mg Vaginal mucosal irritation test and vaginal acute toxicity test were conducted in SD female rats. The stimulation was evaluated by Eckstein irritation evaluation criteria. The results showed that (O-9) - VTG had no effect on vaginal irritation in SD female rats. To sum up, the prepared (O-9) - VTG gelling temperature is suitable, the temperature sensitivity is good, the quality of the preparation is stable, it has a certain sustained-release effect, the retention time in vivo is long, the tissue compatibility is good, the preparation is safer, more effective, and more convenient to use. The use of contraceptives also provides a basis for further development and research of octylene polyol vaginal topical preparations.
【学位授予单位】：湖北中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943

【参考文献】