Folate analogues. 32. Synthesis and biological evaluation of 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolic acid and related compounds.
Folate analogues. 32. Synthesis and biological evaluation of 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolic acid and related compounds.
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叶酸类似物。
DOI：
10.1021/jm00126a023
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发表时间：
1989
影响因子：
7.3
通讯作者：
Ferone,R
中科院分区：
文献类型：
--
作者：
Patil,SD;Jones,C;Nair,MG;Galivan,J;Maley,F;Kisliuk,RL;Gaumont,Y;Duch,D;Ferone,R
描述了 5, 8-二脱氮杂叶酸 (PDDF) 的三种相似类似物 (2-4) 的化学合成。 2 和 4 的喹唑啉环是在合成的最后一步在非常温和的条件下以新颖且明确的方式由关键中间体 9 构建的。 2-去氨-2-甲基-。 N10-propargyl-5, 8-dideazafolate (DMPDDF)(2) 是人类和干酪乳杆菌胸苷酸合成酶的强抑制剂，而 2-desamino-2-(三氟甲基)-IV10-propargyl-5, 8-dideazafolate (3) 和 2-desamino-2, 3-二甲基-iV10-propargyl-5, 8-二脱氮杂叶酸 (4) 只是该酶的弱抑制剂。 DMPDDF 对培养的 Manca 人淋巴白血病和 H35 肝癌细胞表现出优异的生长抑制作用。在这些细胞系中，2 的抑制剂活性分别比 PDDF 高 43 倍和 65 倍。由于运输缺陷而对甲氨蝶呤具有抗性的 H35R 细胞对 DMPDDF 具有交叉抗性，但对 PDDF 没有交叉抗性。与 H35N 细胞相比，H35FF 细胞的胸苷酸合酶含量高出 70 倍，对 DMPDDF 的耐药性高出 130 倍。此外，胸苷可以完全逆转 DMPDDF 对 H35 肝癌细胞的毒性，确定其作为胸苷酸合酶的作用位点。体外转运研究表明，DMPDDF 可有效抑制 MTX 流入 H35 肝癌细胞，而 PDDF 对该细胞系中的 MTX 转运没有影响。这些数据表明，DMPDDF 相对于 PDDF 具有更大的活性，部分原因是前一种化合物能够通过 MTX/还原叶酸转运系统进入细胞。 4 的酶抑制数据表明，DMPDDF 中 N3H 的存在对于与胸苷酸合酶的结合至关重要。Ah-propargyl-5, 8-dideaza-folate (PDDF) 作为胸苷酸合酶 (TS, EC 2.1. 1.45) 特异性抑制剂的显着效力，加上其在小鼠肿瘤模型中作为抗白血病剂的出色体内活性，促使许多研究人员合成了该先导化合物的几种类似物作为潜在的抗癌剂。 3-6 然而，PDDF 的临床实用性因其不溶性和随后的肾毒性而受到影响。 7 因此，PDDF 已退出进一步的临床试验。与叶酸和许多抗叶酸剂一样，PDDF 在体外和体内均已被证明是叶酰聚谷氨酸合成酶 (FPGS) 的底物。 9 PDDF 的多谷氨酰代谢物已在动物 9 和培养的肿瘤细胞中得到鉴定，10, 11 主要形式是四谷氨酸或更高形式。 9, 10 我们之前已经证明，PDDF 的合成聚谷氨酰衍生物比 PDDF 对人类干酪乳杆菌和 L1210 胸苷酸合酶具有更强的抑制作用。 12, 13 根据这些数据，可以合理地假设，与甲氨蝶呤一样，PDDF 的多谷氨酰化是 PDDF 细胞毒性的决定因素。因此，能够增强流入的 PDDF 类似物
The chemical synthesis of three close analogues (2-4) of A^-propargyl-5, 8-dideazafolate (PDDF) is described. The quinazoline ring of 2 and 4 was constructed from the pivotal intermediate 9 in a novel and unambiguous manner during the final step of the synthesis under very mild conditions. 2-Desammo-2-methyl-. N10-propargyl-5, 8-dideazafolate (DMPDDF)(2) was a strong inhibitor of human and Lactobacillus casei thymidylate synthases, whereas 2-desamino-2-(trifluoromethyl)-IV10-propargyl-5, 8-dideazafolate (3) and 2-desamino-2, 3-dimethyl-iV10-propargyl-5, 8-dideazafolate (4) were only weak inhibitors of this enzyme. DMPDDF exhibitedexcellent growth inhibition of Manca human lymphoid leukemia and H35 hepatoma cells in culture. The inhibitor activities of 2 were 43-and65-fold greater thanthat of PDDF, respectively, in these cell lines. H35R cells that are resistant to methotrexate by virtue of a transport defect were cross resistant to DMPDDF but not to PDDF. H35FF cells which have 70-fold greater amounts of thymidylate synthase compared to H35N cells were 130-fold resistant to DMPDDF. Furthermore, the toxicity of DMPDDF to H35 hepatoma cells could be completely reversed by thymidine, establishing its locus of action as thymidylate synthase. Trasnport studies in vitro established that DMPDDF effectively inhibits MTX influx into H35 hepatoma cells, whereas PDDF has no effect on MTX transport in thiscell line. These data suggest that the greater activity of DMPDDF relative to PDDF is partly due to the ability of the former compound to enter cells via the MTX/reduced folate transport system. Enzyme inhibition data of 4 suggest that the presence of N3H in DMPDDF is essential for binding to thymidylate synthase.The remarkable potency of Ah-propargyl-5, 8-dideaza-folate (PDDF) as a specific inhibitor of the enzyme thymidylate synthase (TS, EC 2.1. 1.45) coupled with its ex-cellent in vivo activity as an antileukemic agent in murine tumor models2 has prompted many investigators to synthesize several analogues of this lead compound as po-tential anticancer agents. 3-6 However, the clinical utility of PDDF has suffered due to its insolubility and subse-quent nephrotoxicity. 7 Consequently PDDF has been withdrawn from further clinical trials. Like folate, and many antifolates, PDDF has been shownto be a substrate of folylpolyglutamatesynthetase (FPGS) both in vitro8 and in vivo. 9 Polyglutamyl metabolites of PDDF have been identified in animals9 and tumor cells in culture, 10, 11 the predominant form being the tetraglutamate or higher. 9, 10 We have demonstrated previously that the synthetic po-lyglutamyl derivatives of PDDF were more inhibitory to human, Lactobacillus casei, and L1210 thymidylate syn-thases than PDDF. 12, 13 From these data, it was reasonable to assume that like methotrexate, polyglutamylation of PDDF is a determinant of PDDF cytotoxicity. Therefore, analogues of PDDF that are capable of enhanced influx