利用固相合成法合成了LyP-1，一种对肿瘤细胞具有靶向能力的环状九肽，及其荧光素标记物(LyP-1-FAM)。利用巯基和马来酰亚胺的专属性反应制备了功能化脂质材料LyP-1-PEG 3400-DSPE。用成膜水化法制备了LyP-1 修饰的多柔比星(1)、荧光素(FAM) 和近红外染料(DiR) 脂质体，并评价其对SCI 375 黑素瘤细胞的体内外靶向性、细胞毒性和体内抑瘤效果。体外试验表明，SCI 375 细胞对LyP-1-FAM 或LyP-1 修饰的FAM 脂质体的摄取显著高于5-FAM 或普通FAM 脂质体。LyP-1 修饰及未修饰的DiR 脂质体分别尾静脉注射给予荷瘤裸鼠后，可见LyP-1 修饰组肿瘤组织的荧光强度较高，提示DiR 脂质体经LyP-1 修饰后体内靶向性提高。LyP-1 修饰及未修饰的1 脂质体在体外对SCI 375 细胞的IC50分别为3.4×10-6 和8.0×10-6 mol/L；修饰组在荷瘤裸鼠体内的抑瘤效果也显著高于未修饰组(P<0.05)。
Abstract
LyP-1, a cyclic nonapeptide with the function of tumor targeting, and its fluorescence labeled derivative (LyP-1-FAM) were synthesized by a solid-phase synthesis method. Functional lipid material, LyP-1-PEG 3400-DSPE, was synthesized by the specific reaction between sulfydryl and maleimide group. LyP-1-conjugated liposomes of doxorubicin (1), fluorescein (FAM) and DiR (a near-infrared fluorescent cyanine dye) were prepared by a film hydration method, respectively. The in vitro and in vivo targeting effects and anti-tumor effects on SCI 375 melanoma cells were investigated. The results showed that the in vitro cellular uptake of LyP-1-FAM or LyP-1-conjugated liposomes loaded with FAM (LyP-1-LS/FAM) was higher than 5-FAM or common liposomes of FAM (LS/FAM), respectively. After iv injection of LyP-1-conjugated liposomes of DiR (LyP-1-LS/DiR) or common liposomes of DiR (LS/DiR) to tumor bearing nude mice, the tumor fluorescence intensity of LyP-1-LS/DiR group was higher than LS/DiR group, which indicated that the targeting effect was improved after the modification of LyP-1. The IC50 values of LyP-1-conjugated liposomes of 1 (LyP-1-LS/1) and liposomes of 1 (LS/1) for SCI 375 cells were 3.4×10-6 and 8.0×10-6 mol/L, respectively. The tumor growth inhibition effect of LyP-1-LS/1 in tumor bearing nude mice was more significant than LS/1 (P<0.05).
关键词
LyP-1 /
脂质体 /
多柔比星 /
肿瘤靶向 /
肿瘤治疗
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Key words
LyP-1 /
liposome /
doxorubicin /
tumor targeting /
tumor therapy
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脚注
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基金
国家自然科学基金(81072593)、教育部高等学校博士学科点专项科研基金(20110071130011)、国家重大新药创制科技专项(2012ZX09304004)
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