不同链长脂肪酸修饰对康普瑞汀前药脂质体体内外性质的影响

发布时间：2018-05-09 11:33

本文选题：康普瑞汀脂肪酸酯 + 脂质体　；参考：《第二军医大学》2017年硕士论文

【摘要】：药物的释放对其疗效的提高以及毒副作用的降低具有很大影响,特别是母药从前药中的释放或者转化对其活性药物的药效发挥有着至关重要的作用。本研究选择新型抗肿瘤药物康普瑞汀(Combretastatin A4,CA4)作为模型药物,通过将其制成一系列前药,来研究前药的释放或转化速率对其活性药物体内外性质的影响,从而为筛选出可以提高康普瑞汀抗肿瘤活性的前药制剂打下基础。本文研究内容分为五个部分:(1)康普瑞汀脂肪酸酯的合成与处方前研究,包括康普瑞汀不同链长脂肪酸酯化合物的合成,采用不同方法对合成的化合物进行结构鉴定以及处方前研究;(2)康普瑞汀脂肪酸酯脂质体的制备与表征,包括康普瑞汀脂肪酸酯脂质体处方优化以及脂质体的释放;(3)康普瑞汀脂肪酸酯脂质体体外性质研究,包括血浆中稳定性和体外细胞毒性考查;(4)康普瑞汀脂肪酸酯脂质体体内药代动力学特征和组织分布研究;(5)康普瑞汀脂肪酸酯脂质体体内药效学研究和安全性评价。第一部分,通过脂肪酰氯与康普瑞汀直接进行一步酯化反应生成康普瑞汀脂肪酸酯。通过低分辨质谱、高分辨质谱、核磁共振氢谱和核磁共振碳谱对合成的化合物进行结构鉴定,说明康普瑞汀脂肪酸酯成功合成。并且对康普瑞汀脂肪酸酯进行处方前研究。第二部分,采用薄膜分散法制备康普瑞汀脂肪酸酯脂质体,即康普瑞汀己酸酯脂质体(CA4-6-L)、康普瑞汀癸酸酯脂质体(CA4-10-L)、康普瑞汀肉豆蔻酸酯脂质体(CA4-14-L)、康普瑞汀棕榈酸酯脂质体(CA4-16-L)、康普瑞汀硬酯酸酯脂质体(CA4-6-L)。通过单因素实验考察不同DSPE-PEG2000、不同药脂比和不同磷脂与胆固醇之比对康普瑞汀脂肪酸酯脂质体粒径、PDI、Zeta电位和包封率的影响,并筛选出最终处方为:DSPE-PEG2000用量为0.2%,磷脂与药物之比为9:1,磷脂与胆固醇之比为9:1。通过反向透析法考察康普瑞汀脂质体在体外的释放行为,结果表明随着修饰CA4的脂肪链的增长,前药在体外释放越缓慢。第三部分,首先考察康普瑞汀脂肪酸酯脂质体在大鼠血浆中的稳定性,即前药转化为母药的速度,研究表明,在体外释放越慢的康普瑞汀脂肪酸酯脂质体在血浆中转化为母药的速度也越慢。其次通过CCK8法,对康普瑞汀脂肪酸酯脂质体体外细胞毒性进行考察,细胞毒性结果为CA4-6-LCA4-10-LCA4-14-LCA4-16-LCA4-18-L,即在体外释放越慢,在血浆中转化为母药越慢,其体外细胞毒性越小。第四部分,采用HPLC分析CA4-6-L、CA4-10-L、CA4-18-L和CA4P大鼠体内药代动力学特征,发现活性药物释放慢的CA4-18-L能够延缓活性药物CA4的清除,而在体外释放较快的康普瑞汀脂肪酸酯脂质体在体内则极易被清除。与其他给药组相比,CA4-18-L能够极大的增加CA4的T1/2和AUC,T1/2是其他组的2-4倍,AUC约是其他组的9倍。建立荷S180昆明小鼠模型,评价CA4-10-L、CA4-18-L和CA4P在体内的分布情况,表明CA4-18-L可以增加CA4肿瘤部位的药物浓度。第五部分,首先对CA4P溶液、CA4-10-L和CA4-18-L在荷瘤昆明小鼠体内抗肿瘤活性进行研究,结果显示在相同剂量下CA4P溶液、CA4-10-L和CA4-18-L的抑瘤率分别为57.87%,43.06%和94.44%,进一步证明药物的释放、以及母药从前药中的释放对药物的体内性质有显著影响,释放慢的药物,体内滞留时间长,体内药效好。其次也发现CA4-18-L的体内药效显著好于正处于临床研究的CA4P。另外考察了CA4-18-L的体外溶血性和刺激性,结果显示CA4-18-L未出现溶血现象和刺激性反应。
[Abstract]:The release of drugs has a great effect on the improvement of its efficacy and the decrease in side effects, especially the release or transformation of the maternal antidrug to its active drug effect. This study chose the new antitumor drug Combretastatin A4 (CA4) as a model drug and made it by making it. A series of prodrugs to study the effect of the release or transformation rate of the prodrug on the internal and external properties of its active drug, which is the basis for screening the antineoplastic agents that can improve the antineoplastic activity of coprine. This study is divided into five parts: (1) the synthesis and pre prescription study of the fatty acid esters of coprine, including the different coprine Synthesis of chain long fatty acid esters, structural identification of synthetic compounds and pre formulation studies by different methods; (2) preparation and characterization of coprine fatty acid ester liposomes, including optimization of liposome formulation and release of liposomes; (3) in vitro research on the liposomes of coprine fatty acid ester liposomes It includes plasma stability and cytotoxicity in vitro; (4) pharmacokinetic characteristics and tissue distribution of corpine liposome in liposomes; (5) pharmacodynamic study and safety evaluation of corpine liposome in vivo. The first part is a direct esterification of fatty acid chloride and corprine. The synthesis of compound was identified by low resolution mass spectrometry, high resolution mass spectrometry, nuclear magnetic resonance and magnetic resonance carbon spectroscopy. The successful synthesis of kangpristin fatty acid ester. And the pre - prescription study of the kangpristin fatty acid ester. Second parts were prepared by thin film dispersion method. Ester liposomes, namely, CA4-6-L, CA4-10-L, CA4-10-L, CA4-14-L, CA4-10-L, and CA4-6-L liposome of connecticuline palmitate (CA4-6-L). The effects of different phospholipids and cholesterol on the particle size, PDI, Zeta potential and encapsulation efficiency of the liposome liposome were compared, and the final prescription was selected as: the dosage of DSPE-PEG2000 was 0.2%, the ratio of phospholipid to drug was 9:1, and the ratio of phospholipid to cholesterol was 9:1. through reverse dialysis to investigate the release behavior of the liposome in vitro. The results show that the release of the prodrug in vitro is slower with the increase of the fat chain of the modified CA4. The third part, first inspects the stability of the liposome in the rat plasma, that is, the rate of the prodrug conversion into the mother drug. The study shows that the slower the release of the liposome of the propretine fatty acid ester in vitro is converted into a mother drug in the plasma. The slower the cytotoxicity of the liposome in vitro was examined by the CCK8 method. The cytotoxicity of the liposomes was CA4-6-LCA4-10-LCA4-14-LCA4-16-LCA4-18-L, the slower the release in vitro, the slower the plasma in the plasma, the smaller the cytotoxicity in vitro. The fourth part, using HPLC to analyze CA4-6-L, CA4-10-L The pharmacokinetic characteristics of CA4-18-L and CA4P rats showed that the slow release of CA4-18-L from the active drug delayed the removal of the active drug CA4, while the fast releasing conopertin fatty acid ester liposomes were easily removed in the body. Compared with other drug groups, CA4-18-L could greatly increase the T1/2 and AUC of CA4, T1/2 is the other. 2-4 times of the group, AUC is about 9 times that of the other groups. Establish a S180 Kunming mouse model, evaluate the distribution of CA4-10-L, CA4-18-L and CA4P in the body, indicating that CA4-18-L can increase the concentration of drug in the CA4 tumor site. Fifth, first of all, CA4P solution, CA4-10-L and CA4-18-L in the tumor bearing Kunming mice anti-tumor activity in the body, the results showed that The inhibition rates of CA4P solution at the same dose, CA4-10-L and CA4-18-L were 57.87%, 43.06% and 94.44%, respectively. The release of drugs was further demonstrated, and the release of the drug in the maternal drug had a significant effect on the properties of the drug in vivo, the slow release of the drug, the long retention time in the body and the good efficacy in the body. Secondly, the efficacy of CA4-18-L in vivo was also found. Yu Zheng, who was in the CA4P. clinical study, also examined the hemolysis and irritation of CA4-18-L in vitro. The results showed that CA4-18-L did not appear hemolysis and irritation.

【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943

【参考文献】