Department of Pharmaceutical Informatics, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Department of Chemistry of Functional Molecules, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Int J Nanomedicine. 2022 Dec 28;17:6675-6686. doi: 10.2147/IJN.S390866. eCollection 2022.
Targeted liposomes using ligand peptides have been applied to deliver therapeutic agents to the target sites. The post-insertion method is commonly used because targeted liposomes can be prepared by simple mixing of ligand peptide-lipid and liposomes. A large-scale preparation method is required for the clinical application of ligand-peptide-modified liposomes. Large-scale preparation involves an increase in volume and a change in the preparation conditions. Therefore, the physicochemical properties of liposomes may change owing to large alterations in the preparation conditions. To address this issue, we focused on a microfluidic device and developed a novel ligand peptide modification method, the microfluidic post-insertion method.
We used integrin αvβ3-targeted GRGDS (RGD) and cyclic RGDfK (cRGD)-modified high functionality and quality (HFQ) lipids, which we had previously developed. First, the preparation conditions of the total flow rate in the microfluidic device for modifying HFQ lipids to polyethylene glycol (PEG)-modified (PEGylated) liposomes were optimized by evaluating the physicochemical properties of the liposomes. The targeting ability of integrin αvβ3-expressing colon 26 murine colorectal carcinoma cells was evaluated by comparing the cellular association properties of the liposomes prepared by the conventional post-insertion method.
When the RGD-HFQ lipid was modified into PEGylated liposomes by varying the total flow rate (1, 6, and 12 mL/min) of the microfluidic device, as the total flow rate increased, the polydispersity index also increased, whereas the particle size did not change. Furthermore, the RGD- and cRGD-modified PEGylated liposomes prepared at a total flow rate of 1 mL/min showed high cellular association properties equivalent to those prepared by the conventional post-insertion method.
Microfluidic post-insertion method of HFQ lipids might be useful for clinical application and large-scale preparation of targeted liposomes.
靶向脂质体使用配体肽已被应用于将治疗剂递送到靶部位。后插入方法通常用于因为靶向脂质体可以通过配体肽-脂质和脂质体的简单混合来制备。对于配体肽修饰的脂质体的临床应用需要大规模的制备方法。大规模制备涉及体积的增加和制备条件的变化。因此，由于制备条件的大变化，脂质体的物理化学性质可能会发生变化。为了解决这个问题，我们专注于微流控装置，并开发了一种新的配体肽修饰方法，即微流控后插入方法。
我们使用了整合素 αvβ3 靶向 GRGDS（RGD）和环状 RGDfK（cRGD）修饰的高功能和高质量（HFQ）脂质，这是我们之前开发的。首先，通过评估脂质体的物理化学性质，优化了微流控装置中用于修饰聚乙二醇（PEG）修饰（PEGylated）脂质体的总流速的制备条件。通过比较通过传统后插入法制备的脂质体的细胞结合特性，评估了整合素 αvβ3 表达结肠 26 鼠结直肠癌细胞的靶向能力。
当 RGD-HFQ 脂质通过改变微流控装置的总流速（1、6 和 12 mL/min）来修饰成 PEGylated 脂质体时，随着总流速的增加，多分散指数也增加，而粒径没有变化。此外，在总流速为 1 mL/min 时制备的 RGD 和 cRGD 修饰的 PEGylated 脂质体表现出与传统后插入法制备的相当高的细胞结合特性。
HFQ 脂质的微流控后插入方法可能对靶向脂质体的临床应用和大规模制备有用。
