Division of Pharmacy and Optometry, Faculty of Biology, Medicine and Health, University of Manchester, Manchester Academic Health Science Centre, Oxford Road, Manchester M13 9PL, UK.
Division of Pharmacy and Optometry, Faculty of Biology, Medicine and Health, University of Manchester, Manchester Academic Health Science Centre, Oxford Road, Manchester M13 9PL, UK; Maternal and Fetal Health Research Centre, Division of Developmental Biology and Medicine, Faculty of Biology, Medicine and Health, University of Manchester, 5th floor (Research), St Mary's Hospital, Oxford Road, Manchester M13 9WL, UK; St Mary's Hospital, Manchester University NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester M13 9WL, UK.
Int J Pharm. 2020 Nov 30;590:119926. doi: 10.1016/j.ijpharm.2020.119926. Epub 2020 Oct 1.
Developing more efficient manufacturing methods for nano therapeutic systems is becoming important, not only to better control their physico-chemical characteristics and therapeutic efficacy but also to ensure scale-up is cost-effective. The principle of cross-flow chemistry allows precise control over manufacturing parameters for the fabrication of uniform liposomal formulations, as well as providing reproducible manufacturing scale-up compared to conventional methods. We have herein investigated the use of microfluidics to produce PEGylated DSPC liposomes loaded with doxorubicin and compared their performance against identical formulations prepared by the thin-film method. The isoprenylated coumarin umbelliprenin was selected as a co-therapeutic. Umbelliprenin-loaded and doxorubicin:umbelliprenin co-loaded liposomes were fabricated using the optimised microfluidic set-up. The role of umbelliprenin as lipid bilayer fluidity modulation was characterized, and we investigated its role on liposomes size, size distribution, shape and stability compared to doxorubicin-loaded liposomes. Finally, the toxicity of all liposomal formulations was tested on a panel of human breast cancer cells (MCF-7, MDA-MB 231, BT-474) to identify the most potent formulation by liposomal fabrication method and loaded compound(s). We herein show that the microfluidic system is an alternative method to produce doxorubicin:umbelliprenin co-loaded liposomes, allowing fine control over liposome size (100-250 nm), shape, uniformity and doxorubicin drug loading (>80%). Umbelliprenin was shown to confer fluidity to model lipid biomembranes, which helps to explain the more homogeneous size and shape of co-loaded liposomes compared to liposomes without umbelliprenin. The toxicity of doxorubicin:umbelliprenin co-loaded liposomes was lower than that of free doxorubicin, due to the delayed release of doxorubicin from liposomes. An alternative, rapid and easy manufacturing method for the production of liposomes has been established using microfluidics to effectively produce uniform doxorubicin:umbelliprenin co-loaded liposomal formulations with proven cytotoxicity in human breast cancer cell lines in vitro.
开发更有效的纳米治疗系统制造方法变得越来越重要，不仅可以更好地控制其物理化学特性和治疗效果，还可以确保扩大规模具有成本效益。 逆流化学的原理允许对制造参数进行精确控制，从而制造出均匀的脂质体配方，并且与传统方法相比，还可以提供可重复的制造扩大规模。 本文研究了使用微流控技术生产载有多柔比星的 PEG 化 DSPC 脂质体，并将其性能与通过薄膜法制备的相同配方进行了比较。 异戊二烯化香豆素 umbelliprenin 被选为辅助治疗药物。 用优化的微流控装置制备了载有 umbelliprenin 和载有多柔比星：umbelliprenin 的脂质体。 表征了 umbelliprenin 作为脂质双层流动性调节剂的作用，并研究了其对脂质体大小，大小分布，形状和稳定性的影响，与载有多柔比星的脂质体相比。 最后，通过脂质体制备方法和负载的化合物，用人乳腺癌细胞（MCF-7、MDA-MB 231、BT-474）测试了所有脂质体配方的毒性，以鉴定最有效的配方。 本文表明，微流控系统是生产多柔比星：umbelliprenin 共载脂质体的替代方法，可以精细控制脂质体大小（100-250nm）、形状、均匀性和多柔比星药物载量（>80%）。 umbelliprenin 被证明可以赋予模型脂质生物膜流动性，这有助于解释与不含 umbelliprenin 的脂质体相比，共载脂质体的尺寸和形状更加均匀。 由于多柔比星从脂质体中的释放延迟，多柔比星：umbelliprenin 共载脂质体的毒性低于游离多柔比星。 已经建立了一种替代的、快速且易于使用的制造方法，用于使用微流控技术生产均匀的多柔比星：umbelliprenin 共载脂质体配方，该配方在体外对人乳腺癌细胞系具有证明的细胞毒性。
