Shiokawa Tomonori, Hattori Yoshiyuki, Kawano Kumi, Ohguchi Yukino, Kawakami Hiroko, Toma Kazunori, Maitani Yoshie
Institute of Medicinal Chemistry, Hoshi University, Ebara 2-4-41, Shinagawa-ku, Tokyo 142-8501, Japan.
Clin Cancer Res. 2005 Mar 1;11(5):2018-25. doi: 10.1158/1078-0432.CCR-04-1129.
To establish a novel formulation tumor-targeted drug carrier of lipophilic antitumor antibiotics, aclacinomycin A (ACM), folate-linked microemulsions were prepared and investigated both in vitro and in vivo.
Three kinds of folate-linked microemulsions with different polyethylene glycol (PEG) chain lengths loading ACM were formulated with 0.24 mol% folate-PEG(2000)-distearoylphosphatidylethanolamine (DSPE), folate-PEG(5000)-DSPE, and folate-lipid (without PEG linker) in microemulsions. In vitro studies were done in a human nasopharyngeal cell line, KB, which overexpresses the folate receptor (FR), and a human hepatoblastoma cell line, [FR(-)] HepG2. In vivo experiments were done in a KB xenograft by systemic administration of folate-linked microemulsions loading ACM.
The association of folate-linked microemulsions to KB cells could be blocked by 2 mmol/L free folic acid. Selective FR-mediated cytotoxicity of folate-linked microemulsions loading ACM was obtained in KB but not in HepG2 cells. The association of the folate-PEG(5000)-linked microemulsion and folate-PEG(2000)-linked microemulsion with the cells was 200- and 4-fold higher, whereas their cytotoxicity was 90- and 3.5-fold higher than those of nonfolate microemulsion, respectively. The folate-PEG(5000)-linked microemulsions showed 2.6-fold higher accumulation in solid tumors 24 hours after i.v. injection and greater tumor growth inhibition than free ACM.
These findings suggest that a folate-linked microemulsion is feasible for tumor-targeted ACM delivery. This study shows that folate modification with a sufficiently long PEG chain on emulsions is an effective way of targeting emulsion to tumor cells.
为建立一种新型的亲脂性抗肿瘤抗生素阿克拉霉素A(ACM)的肿瘤靶向药物载体,制备了叶酸连接的微乳剂,并进行了体外和体内研究。
在微乳剂中用0.24 mol%的叶酸-聚乙二醇(PEG)(2000)-二硬脂酰磷脂酰乙醇胺(DSPE)、叶酸-PEG(5000)-DSPE和叶酸-脂质(无PEG连接子)制备了三种负载ACM且具有不同PEG链长度的叶酸连接微乳剂。体外研究在过表达叶酸受体(FR)的人鼻咽癌细胞系KB和人肝癌细胞系[FR(-)]HepG2中进行。体内实验通过全身给药负载ACM的叶酸连接微乳剂在KB异种移植瘤中进行。
2 mmol/L游离叶酸可阻断叶酸连接微乳剂与KB细胞的结合。负载ACM的叶酸连接微乳剂在KB细胞中获得了选择性的FR介导的细胞毒性,而在HepG2细胞中未获得。叶酸-PEG(5000)连接的微乳剂和叶酸-PEG(2000)连接的微乳剂与细胞的结合分别比非叶酸微乳剂高200倍和4倍,而它们的细胞毒性分别比非叶酸微乳剂高90倍和3.5倍。静脉注射后24小时,叶酸-PEG(5000)连接的微乳剂在实体瘤中的蓄积比游离ACM高2.6倍,且对肿瘤生长的抑制作用更强。
这些发现表明叶酸连接微乳剂用于肿瘤靶向递送ACM是可行的。本研究表明在乳剂上用足够长的PEG链进行叶酸修饰是将乳剂靶向肿瘤细胞的有效方法。