【作者】 陈丹；

【导师】 龚涛；

【作者基本信息】 四川大学 ， 药剂学， 2021， 硕士

【摘要】 脑胶质瘤是一种侵袭性的原发恶性脑肿瘤,很难治疗。肝细胞癌是另一种常见的恶性肿瘤,是全球第二大致死癌症。虽然随着药物传递系统和生物制剂的发展,有了越来越多的治疗策略,但化疗仍是治疗脑胶质瘤和肝癌的重要方法之一。细胞毒性化疗药物(如替莫唑胺、紫杉醇和阿霉素)通常会引起严重的不良反应,如骨髓抑制。绿原酸是来源于植物的天然产物,有良好的抗癌活性和低毒性。然而,由于其体内停留时间短、需要长期频繁给药,其应用受到限制。因此,我们考虑将绿原酸载入原位凝胶系统,希望该缓释制剂在治疗脑胶质瘤、肝细胞癌的同时,有较好的安全性和更高的应用价值。本课题对绿原酸原位凝胶进行了较全面的实验研究。首先,建立了绿原酸体外样品与血浆样品的含量测定方法,分别进行了方法学验证。分别选用准确可行的高效液相系统,以及灵敏度高、准确度好的液相色谱串联质谱检测系统作为药物含量的分析测定方法。之后,确定了绿原酸原位凝胶的处方,通过简单的搅拌混合法制得绿原酸原位凝胶,并对绿原酸原位凝胶进行了表征及稳定性研究。绿原酸原位凝胶均一澄明,且流动性及稳定性良好。此外,绿原酸原位凝胶在相变前黏度较低,可注射使用;相变后,绿原酸原位凝胶黏度迅速增大,呈半固体凝胶状。接着,进行了绿原酸原位凝胶的体外释放实验。考察在不同释放介质中,绿原酸原位凝胶的体外释药行为,初步评价该凝胶的缓释情况。与游离绿原酸组相比,绿原酸原位凝胶组药物的体外释放速度明显减慢,释放时间延长。随后,考察了绿原酸原位凝胶在大鼠体内的药动行为,评价了该制剂作为药物储库的缓释能力。结果显示,与游离绿原酸相比,绿原酸原位凝胶能显著延长绿原酸在体内的平均滞留时间及半衰期,体内药物浓度更为稳定,且突释更小,更符合临床治疗的需求。证实了绿原酸原位凝胶良好的缓释作用。然后,考察了绿原酸原位凝胶的体内药效。分别构建了脑胶质瘤、H22肝癌细胞移植瘤小鼠模型。结果表明,相比于游离绿原酸,绿原酸原位凝胶对以上两种荷瘤小鼠模型,均有更好的治疗效果。此外,通过苏木精-伊红(H&E)染色、免疫荧光、免疫组化,初步考察了抗肿瘤机制。最后,对绿原酸原位凝胶进行了初步安全性评价,绿原酸原位凝胶有较好的系统安全性。综上所述,本课题制备了绿原酸原位缓释凝胶,并对其理化性质、稳定性、体外释放行为、体内药动、体内药效及其作用机制、安全性进行了研究。绿原酸原位凝胶具有良好的流动性及稳定性、易注射性、高载药量和适宜的体内外缓释效果。与游离绿原酸相比,其缓释行为更符合临床需要,且安全性好、制备工艺简便,有一定的应用价值。绿原酸原位凝胶对脑胶质瘤和肝细胞癌有良好的治疗作用,而无明显毒副作用,为绿原酸的应用提供了新思路。更多还原

【Abstract】 Glioma is one kind of aggressive primary malignant brain tumors that is very difficult to be treated.Hepatocellular carcinoma(HCC)is another common malignancy which is the second leading cause of cancer-related deaths worldwide.Although more effective treatment strategies are available with progress on drug delivery systems and biological agents,chemotherapy is still one of the most important methods to treat glioma and HCC.Unfortunately,cytotoxic chemotherapeutic agents such as temozolomide,paclitaxel and doxorubicin usually cause serious adverse reactions including bone marrow suppression,liver and kidney function disorder.Notably,chlorogenic acid(CGA)may provide an effective and safe option for tumor treatment.CGA is a therapeutic drug for glioma and H22 transplanted tumor with good safety.However,its application is limited because of short residence time and repeated administration required and high hydrophilicity.A novel approach is the local injectable drug delivery system with sustained release,but there were obstacles,for example,complicated formulation preparation and potential toxicity of excipients.Herein,to realize the safe and efficient delivery of CGA for antitumor therapy,a phospholipid-based in situ forming gel containing chlorogenic acid(CGA-PG)was designed.The CGA-PG exhibited good fluidity,easy injectability,high-drug-loading capacity.And CGA-PG has good storage stability which provided a basis for industrial production.The CGA-PG exhibited suitable sustained-release behavior whether in vitro or in vivo.In vitro release profiles of CGA-PG were studied in different release mediums.Compared with free CGA,CGA-PG showed a slow and sustained release.In vivo pharmacokinetics of CGA-PG were studied in rats.CGA-PG prolonged the residence time of CGA in vivo,and showed a less significant burst phase compared with free CGA.Furthermore,CGA-PG could suppress tumor growth with no significant side effects.CGA-PG was demonstrated to show antitumor efficacy in both a glioma model and a xenograft mouse model of H22.The antitumor efficacy of CGA was probably related to the anti-angiogenic effects,induction of apoptosis,inhibition of the invasion of cancer cells.Moreover,the safety evaluation research of CGA-PG is consisted of three parts:the tolerability of injection site,blood routine evaluation and biochemical examination.CGA-PG showed good safety.In a word,CGA-PG may be a promising sustained drug preparation with excellent therapeutic effect on glioma and H22 transplanted tumor.We feel like our research results will provide new ideas for the application of CGA.更多还原