肝靶向斑蝥素固体脂质纳米粒的研究
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摘要
目的：将斑蝥素制成斑蝥素固体脂质纳米粒(CTD-SLN),从而使其具有缓释、毒性降低、靶向性等优点,在肝癌局部达到减毒增效的目的。
方法：采用薄膜分散-高压微射流法制备CTD-SLN,以CTD-SLN形态、包封率、载药量做为评价指标,综合评分法对处方和制备工艺进行优化；选用低温超速离心、透射电镜观察、激光散射粒度分析、Zeta电位测定对CTD-SLN进行外观形态、粒径、粒度分布等性质进行考察；通过反相高效液相色谱法测定其包封率和载药量；动态透析膜法考察其体外释药特性；考察不同温度、光照等条件下CTD-SLN的稳定性,综合各考察结果对CTD-SLN进行综合评价。以小鼠为模型动物,以斑蝥素水针剂为对照,研究纳米粒的毒性,并比较同等剂量注射时小鼠的临床表现,考察固体脂质纳米粒的减毒效果。
结果：对CTD-SLN制备过程中有显著影响的四种主要因素为：药物-硬脂酸比、磷脂量、泊洛沙姆188量、油-水体积比,根据优化处方与工艺制得三批CTD-SLN,在透射电镜下观察,分散性好、圆整、均匀而不粘连,平均粒径110.4 nm,符合被动肝靶向要求,Zeta电位-23.63 mv,具有一定稳定性。包封率83.6%,载药量3.58%。体外释放规律符合双相动力学过程,其特点为前期为快速释药,后期缓慢释药,这种释药特征比较符合抗癌药物的用药原则,即纳米粒到达靶部位后可立即释放出较多药物,达到较高浓度(冲击量),然后缓慢释出剩余药物,维持较长时间(维持量),可达到缓释长效目的。CTD-SLN的物理稳定性表明,不可耐受加热,适宜低温保存。初步药理学试验表明,纳米粒具有减毒作用。
结论：将斑蝥素制成载药固体脂质纳米粒,可使药物有效地靶向输送于肝脏病变部位、降低其毒副作用,缓释作用明显。本文所涉及的试验方法和理论为中药提取类药物的制剂开发和纳米化研究提供了一定的实验基础。
OBJECTIVE:Cantharidin is made of soLid Lipid nanoparticLes, making it a sLow-reLease, toxicity reduction, targeting, etc., in the Liver to reach attenuated partiaL efficiency purposes.
METHODS:Using High-pressure microfLudizer, in order to CTD-SLN morphoLogy, encapsuLation efficiency, drug Loading as the evaLuation index, comprehensive scoring pairs of prescription and preparation process optimization. Through the RP-HPLC, Low speed centrifugation, TEM, Laser scattering particLe size anaLysis, Zeta potentiaL measurement methods, temperature osciLLation of its physicaL and chemicaL properties and in vitro reLease characteristics of a comprehensive evaLuation. Taking mice as a modeL animaL, in order to cantharidin as controL of water injection to study the toxicity of nanoparticLes and compared the same dose of injection of cLinicaL manifestations in mice, examining the attenuated effect of soLid Lipid nanoparticLes.
RESULTS:1.A good dispersion was observed under transmission eLectron microscope, whoLeness, even without adhesion.2. The average diameter of 110 nm, Zeta potentiaL, has a certain stabiLity.3. EncapsuLation rate of 83.6%,3.58%drug Loading.4. In vitro reLease pattern fitted weLL with the doubLe-exponentiaL equation with a significant sustained reLease characteristics.5.CTD-SLN of the physicaL stabiLity that can not withstand heat, suitabLe for cryopreservation.6. PreLiminary pharmacoLogicaL tests showed that nanoparticLes with the attenuation.
CONCLUSION:Cantharidin drug-Loaded soLid Lipid nanoparticLes wiLL enabLe the effective targeting of drug deLivery in the Liver Lesions, reducing its toxic side effects, sLow-reLease effect is obvious. This invoLved testing methods and the theory of traditionaL Chinese medicine preparations extracted drugs and nano-technoLogy research deveLopment of some experimentaL basis.
方法：采用薄膜分散-高压微射流法制备CTD-SLN,以CTD-SLN形态、包封率、载药量做为评价指标,综合评分法对处方和制备工艺进行优化；选用低温超速离心、透射电镜观察、激光散射粒度分析、Zeta电位测定对CTD-SLN进行外观形态、粒径、粒度分布等性质进行考察；通过反相高效液相色谱法测定其包封率和载药量；动态透析膜法考察其体外释药特性；考察不同温度、光照等条件下CTD-SLN的稳定性,综合各考察结果对CTD-SLN进行综合评价。以小鼠为模型动物,以斑蝥素水针剂为对照,研究纳米粒的毒性,并比较同等剂量注射时小鼠的临床表现,考察固体脂质纳米粒的减毒效果。
结果：对CTD-SLN制备过程中有显著影响的四种主要因素为：药物-硬脂酸比、磷脂量、泊洛沙姆188量、油-水体积比,根据优化处方与工艺制得三批CTD-SLN,在透射电镜下观察,分散性好、圆整、均匀而不粘连,平均粒径110.4 nm,符合被动肝靶向要求,Zeta电位-23.63 mv,具有一定稳定性。包封率83.6%,载药量3.58%。体外释放规律符合双相动力学过程,其特点为前期为快速释药,后期缓慢释药,这种释药特征比较符合抗癌药物的用药原则,即纳米粒到达靶部位后可立即释放出较多药物,达到较高浓度(冲击量),然后缓慢释出剩余药物,维持较长时间(维持量),可达到缓释长效目的。CTD-SLN的物理稳定性表明,不可耐受加热,适宜低温保存。初步药理学试验表明,纳米粒具有减毒作用。
结论：将斑蝥素制成载药固体脂质纳米粒,可使药物有效地靶向输送于肝脏病变部位、降低其毒副作用,缓释作用明显。本文所涉及的试验方法和理论为中药提取类药物的制剂开发和纳米化研究提供了一定的实验基础。
OBJECTIVE:Cantharidin is made of soLid Lipid nanoparticLes, making it a sLow-reLease, toxicity reduction, targeting, etc., in the Liver to reach attenuated partiaL efficiency purposes.
METHODS:Using High-pressure microfLudizer, in order to CTD-SLN morphoLogy, encapsuLation efficiency, drug Loading as the evaLuation index, comprehensive scoring pairs of prescription and preparation process optimization. Through the RP-HPLC, Low speed centrifugation, TEM, Laser scattering particLe size anaLysis, Zeta potentiaL measurement methods, temperature osciLLation of its physicaL and chemicaL properties and in vitro reLease characteristics of a comprehensive evaLuation. Taking mice as a modeL animaL, in order to cantharidin as controL of water injection to study the toxicity of nanoparticLes and compared the same dose of injection of cLinicaL manifestations in mice, examining the attenuated effect of soLid Lipid nanoparticLes.
RESULTS:1.A good dispersion was observed under transmission eLectron microscope, whoLeness, even without adhesion.2. The average diameter of 110 nm, Zeta potentiaL, has a certain stabiLity.3. EncapsuLation rate of 83.6%,3.58%drug Loading.4. In vitro reLease pattern fitted weLL with the doubLe-exponentiaL equation with a significant sustained reLease characteristics.5.CTD-SLN of the physicaL stabiLity that can not withstand heat, suitabLe for cryopreservation.6. PreLiminary pharmacoLogicaL tests showed that nanoparticLes with the attenuation.
CONCLUSION:Cantharidin drug-Loaded soLid Lipid nanoparticLes wiLL enabLe the effective targeting of drug deLivery in the Liver Lesions, reducing its toxic side effects, sLow-reLease effect is obvious. This invoLved testing methods and the theory of traditionaL Chinese medicine preparations extracted drugs and nano-technoLogy research deveLopment of some experimentaL basis.
引文
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