Anticancer agents coupled to N-(2-hydroxypropyl)methacrylamide copolymers. I. Evaluation of daunomycin and puromycin conjugates in vitro.
Anticancer agents coupled to N-(2-hydroxypropyl)methacrylamide copolymers. I. Evaluation of daunomycin and puromycin conjugates in vitro.
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DOI：
10.1038/bjc.1987.33
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发表时间：
1987-02
影响因子：
8.8
通讯作者：
Kopecek, J
中科院分区：
文献类型：
--
作者：
Duncan, R;Kopeckova-Rejmanova, P;Strohalm, J;Hume, I;Cable, H C;Pohl, J;Lloyd, J B;Kopecek, J
近年来，N-（2-羟丙基）甲基丙烯酰胺（HPMA）共聚物被开发为靶向药物载体。这些可溶性合成聚合物通过胞饮作用被细胞内化，并且它们可以被定制为包括可通过特异性溶酶体酶在细胞内降解的肽基侧链。因此，它们提供了实现抗癌剂的受控细胞内递送的机会。蒽环类抗生素道诺霉素和蛋白质合成抑制剂嘌呤霉素通过几种不同的肽侧链（包括Gly-Gly、Gly-Phe-Leu-Gly和Gly-Phe-Phe-Leu）与HPMA共聚物结合。用分离的溶酶体酶（大鼠肝溶酶体酶或纯化的巯基依赖性溶酶体蛋白酶，组织蛋白酶L和B的混合物）孵育聚合物-药物缀合物显示，在20小时内发生药物的显著释放，超过20%的柔红霉素和超过80%的嘌呤霉素被释放。为了测试其药理活性，将缀合物与小鼠白血病L1210或人淋巴母细胞样白血病CCRF在体外孵育。测试的缀合物都不如游离道诺霉素有效，但它们对L1210显示出不同的毒性，这取决于它们的药物-聚合物键的氨基酸序列。将岩藻糖胺终止侧链纳入HPMA共聚物结构中增加了缀合物对L1210细胞膜的亲和力，并导致毒性增加。与此相反，HPMA-道诺霉素共轭物与岩藻糖胺或不受影响的CCRF细胞一样，但这种细胞系比小鼠白血病更敏感的自由和聚合物结合的道诺霉素。L1210细胞在聚合物结合的柔红霉素中孵育72小时，然后在无药物培养基中以低密度铺板细胞，表明可以选择聚合物结合的药物浓度（184微克/ml）以实现细胞毒性作用。
During recent years N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers have been developed as targetable drug carriers. These soluble synthetic polymers are internalized by cells by pinocytosis and they can be tailor-made to include peptidyl side-chains degradable intracellularly by specific lysosomal enzymes. Thus they provide the opportunity fo achieve controlled intracellular delivery of anticancer agents. The anthracycline antibiotic daunomycin, and protein synthesis inhibitor puromycin, were bound to HPMA copolymers via several different peptide side-chains, including Gly-Gly, Gly-Phe-Leu-Gly and Gly-Phe-Phe-Leu. Incubation of polymer-drug conjugates with isolated lysosomal enzymes (either a mixture of rat liver lysosomal enzymes or purified thiol-dependent lysosomal proteinases, cathepsins L and B) showed that significant release of drug occurred over 20 h, more than 20% of daunomycin and more than 80% of puromycin being liberated. To test their pharmacological activity conjugates were incubated with either the mouse leukaemia L1210, or the human lymphoblastoid leukaemia CCRF in vitro. The conjugates tested were all less effective than free daunomycin, but they showed differential toxicity against L1210 depending on the aminoacid sequence of their drug-polymer linkage. Inclusion of fucosylamine-terminating side-chains into the HPMA copolymer structure increased the affinity of conjugates for the L1210 cell membrane and resulted in increased toxicity. In contrast HPMA-daunomycin conjugates with or without fucosylamine affected CCRF cells equally, but this cell line was more sensitive than the mouse leukaemia to both free and polymer-bound daunomycin. Incubation of L1210 cells in polymer-bound daunomycin for 72 h, followed by plating cells out in low density in drug-free medium, showed that a concentration of polymer-bound drug (184 micrograms ml-1) could be selected to achieve a cytotoxic effect.