Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia.
School of Medical Sciences and Prince of Wales Clinical School, UNSW Sydney, Sydney, NSW 2052, Australia.
Biomacromolecules. 2022 Jun 13;23(6):2374-2387. doi: 10.1021/acs.biomac.2c00143. Epub 2022 May 4.
The synthesis of new amino acid-containing, cell-specific, therapeutically active polymers is presented. Amino acids served as starting material for the preparation of tailored polymers with different amino acids in the side chain. The reversible addition-fragmentation chain-transfer (RAFT) polymerization of acrylate monomers yielded polymers of narrow size distribution ( ≤ 1.3). In particular, glutamate (Glu)-functionalized, zwitterionic polymers revealed a high degree of cytocompatibility and cellular specificity, , showing association to different cancer cell lines, but not with nontumor fibroblasts. Energy-dependent uptake mechanisms were confirmed by means of temperature-dependent cellular uptake experiments as well as localization of the polymers in cellular lysosomes determined by confocal laser scanning microscopy (CLSM). The amino acid receptor antagonist -benzyl-l-serine (BzlSer) was chosen as an active ingredient for the design of therapeutic copolymers. RAFT copolymerization of Glu acrylate and BzlSer acrylate resulted in tailored macromolecules with distinct monomer ratios. The targeted, cytotoxic activity of copolymers was demonstrated by means of multiday cell viability assays. To this end, polymers with 25 mol % BzlSer content showed cytotoxicity against cancer cells, while leaving fibroblasts unaffected over a period of 3 days. Our results emphasize the importance of biologically derived materials to be included in synthetic polymers and the potential of zwitterionic, amino acid-derived materials for cellular targeting. Furthermore, it highlights that the fine balance between cellular specificity and unspecific cytotoxicity can be tailored by monomer ratios within a copolymer.
新型含氨基酸、细胞特异性、治疗活性聚合物的合成。氨基酸作为起始原料，用于制备具有不同侧链氨基酸的定制聚合物。丙烯酸酯单体的可逆加成-断裂链转移（RAFT）聚合得到了窄分布的聚合物（≤1.3）。特别是谷氨酸（Glu）功能化的两性离子聚合物表现出高度的细胞相容性和细胞特异性，与不同的癌细胞系结合，但与非肿瘤成纤维细胞不结合。通过依赖能量的细胞摄取实验以及通过共聚焦激光扫描显微镜（CLSM）确定聚合物在细胞溶酶体中的定位，证实了能量依赖的摄取机制。选择氨基酸受体拮抗剂 -苄基-l-丝氨酸（BzlSer）作为设计治疗性共聚物的有效成分。谷氨酸丙烯酸酯和 BzlSer 丙烯酸酯的 RAFT 共聚得到了具有明显单体比的定制大分子。通过多日细胞活力测定，证明了共聚物的靶向细胞毒性活性。为此，含有 25mol%BzlSer 含量的聚合物对癌细胞表现出细胞毒性，而在 3 天的时间内对成纤维细胞没有影响。我们的结果强调了将生物衍生材料纳入合成聚合物中的重要性，以及两性离子、氨基酸衍生材料在细胞靶向方面的潜力。此外，它还强调了通过共聚物中单体比例可以精细调整细胞特异性和非特异性细胞毒性之间的平衡。
