Cellulose and Paper Department, National Research Center, Dokki, 12622 Cairo, Egypt; Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, United States.
Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, United States.
Int J Biol Macromol. 2018 Jun;112:694-702. doi: 10.1016/j.ijbiomac.2018.01.213. Epub 2018 Feb 2.
In this studies, three fatty acyl derivatives of CGKRK homing peptides were coupled successfully to chitosan oligosaccharides (COS) using sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate sodium salt (sulfo-SMCC). The COS-SMCC was prepared by direct coupling between COS and sulfo-SMCC in PBS (pH7.5) at RT for 48h. The structure of COS-SMCC and the three fatty acyl-CGKRK-SMCC-COS conjugates were characterized by FT-IR, C NMR, and SEM. The ability of three conjugates to condense siRNA into nanosized polyplexes and their efficacy in protecting siRNA from serum nucleases degradation were investigated. Among the investigated derivatives, S-CGKRK-COS showed higher siRNA binding affinity as compared to the P-CGKRK-COS and O-CGKRK-COS, respectively. At a ratio of 10:1, complete protection for siRNA from early enzymatic degradation was achieved. The polymers and the polymer/siRNA polyplexes showed negligible cytotoxicity on human breast cancer cell line MDA-MB-231 at all investigated ratios. However, the polyplexes prepared with palmitoyl and oleoyl derivatives at polymer concentration 10μg/mL reduced the cell viability by 21.5% and 35%, respectively. The results of this study revealed the potential use of fatty acyl-CGKRK-COS as a siRNA carrier and confirmed the importance of incorporating a hydrophobic moiety into chitosan to improve its capacity in complexing with siRNA and protection from degradation.
在这项研究中，成功地将三个脂肪酸酰基衍生物的 CGKRK 归巢肽与壳聚糖寡糖（COS）通过磺基琥珀酰亚胺基-4-（N-马来酰亚胺基甲基）环己烷-1-羧酸酯钠盐（sulfo-SMCC）偶联。COS-SMCC 通过 COS 与 sulfo-SMCC 在 PBS（pH7.5）中于室温下直接偶联 48 小时制备。COS-SMCC 和三种脂肪酸酰基-CGKRK-SMCC-COS 缀合物的结构通过傅里叶变换红外光谱（FT-IR）、碳 NMR 和 SEM 进行了表征。研究了三种缀合物将 siRNA 凝聚成纳米级多聚物的能力及其保护 siRNA 免受血清核酶降解的功效。在所研究的衍生物中，S-CGKRK-COS 与 P-CGKRK-COS 和 O-CGKRK-COS 相比，显示出更高的 siRNA 结合亲和力。在 10:1 的比例下，完全保护 siRNA 免受早期酶降解。在所有研究的比例下，聚合物及其聚合物/siRNA 多聚物对人乳腺癌细胞 MDA-MB-231 表现出可忽略不计的细胞毒性。然而，在聚合物浓度为 10μg/mL 时，用棕榈酰和油酰衍生物制备的多聚物使细胞活力分别降低了 21.5%和 35%。这项研究的结果表明，脂肪酸酰基-CGKRK-COS 作为 siRNA 载体具有潜在的应用价值，并证实了将疏水性部分整合到壳聚糖中以提高其与 siRNA 结合的能力并保护其免受降解的重要性。
