包载NLG919的乳腺癌靶向纳米胶束的构建及体外评价

Construction and in vitro evaluation of NLG919-loaded breast cancer-targeting nanomicelles

目的:制备包载NLG919的乳腺癌靶向纳米胶束,以改善NLG919的水溶性,提高靶向性,并对其体外性质进行考察。方法:采用固相合成法合成胆固醇修饰的多肽单体CHL,采用透析法制备NLG919-CHL纳米胶束,并用动态光散射法测定粒径,用芘荧光探针测定其临界胶束浓度,采用激光共聚焦法考察其在细胞内的分布情况,并测定其对小鼠乳腺癌肿瘤细胞4T1和人乳腺癌肿瘤细胞MCF-7的细胞毒性,及对肿瘤细胞吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)活性的抑制作用。结果:采用透析法成功制备了NLG919-CHL纳米胶束,胶束的载药率和包封率分别为(21.04±0.002)%和(63.30±0.011)%。细胞毒性实验结果表明,当空白胶束Blank-CHL浓度达到200μg/ml,作用4T1和MCF-7细胞24 h后,细胞活力仍>85%。激光共聚焦实验结果表明载体可以将药物载送到细胞内。结论:NLG919纳米胶束构建成功,有望成为一种低毒、乳腺癌靶向的药物递送载体。

Objective:To prepare NLG919-loaded breast cancer-targeting nanomicelles,so as to improve water solubility and targeting ability of NLG919,and also to investigate the characteristics of nanomicelles in vitro.Methods:The cholesterol-modified peptide monomer CHL was synthesized by solid phase synthesis.NLG919-CHL nanomicelles were prepared by dialysis method.Particle sizes were measured by dynamic light scattering,and critical micelle concentration was investigated by pyrene fluorescein probe.Laser confocal technique was used to investigate the intracellular distribution of cells.Cytotoxicity of nanomicelles to tumor cells 4T1 of mouse breast cancer and tumor cells MCF-7 of human breast cancer were determined,and activity inhibition of indoleamine 2,3-dioxygenase(IDO)in tumor cells were also investigated.Results:NLG919-CHL nanomicelles were successfully prepared by dialysis method.The drug loading rate and encapsulation efficiency of micelles reached as high as(21.04±0.002)%and(63.30±0.011)%,respectively.The results of cytotoxicity experiment showed that the cell viability of 4T1 and MCF-7 cells was still above 85%when it was cultured with the Blank-CHL at the concentration of 200μg/ml for 24 hours.The results of confocal experiment indicated that the nanomicelles could carry the drug into cells.Conclusion:The successful construction of NLG919-loaded nanomicelles might promise a low-toxic,breast cancer-targeted drug delivery system.