As 17β-estradiol and its derivatives have good affinity with estrogen receptors and have unique antitumor properties, O¹-propargyl-N⁴,N⁴-bis(2-chloroethyl)-4-aminophenol and N⁴,N⁴-dipropargyl-N¹,N¹-bis(2-chloroethyl)-1,4-phenylenediamine were respectively combined with 17β-estradiol through linkers of different lengths. These targeted novel nitrogen mustard 17β-estradiol derivatives were tested antiproliferative activity using the MTT assay. The results illustrated that these compounds were less toxicity to HL-7702 (human liver cells) and SKOV-3, though all their activity potency was moderate or weak aganist the MDA-MB-231, MCF-7 and BEL-7404 cell lines. The inhibitory activity of the O¹-propargyl-N⁴,N⁴-bis (2-chloroethyl)-4-aminophenol derivatives (compounds III-1~6 and VII-1~4) were obviously better than the N⁴-dipropargyl-N¹, N¹-bis (2-chloroethyl) -1,4-phenylenediamine derivatives (compounds IV-1~6 and VIII-1~4) against tested tumor cell lines. And three hybrids (compounds III-2, III-6, and VII-3) exhibited good inhibitory activity against BEL-7404 cell line (IC₅₀= 13.5, 15.3 and 13.9 μM, respectively). After that, we performed KEGG pathway analysis on compound III-6, and the results showed that III-6 mainly induced breast cancer cell apoptosis by regulating the expression of the proteins in NF-κ B pathway.

由于 17 β-雌二醇及其衍生物与雌激素受体有良好的亲和力并具有独特的抗肿瘤特性，O ^{1 -}炔丙基-N ⁴ , N ^{4 -}双(2-氯乙基)-4-氨基苯酚和N ⁴ , N ^{4 -二}炔丙基- N ¹、N ^{1 -}双(2-氯乙基)-1,4-苯二胺分别通过不同长度的接头与17 β-雌二醇结合。这些靶向新型氮芥17β使用 MTT 测定法测试了 - 雌二醇衍生物的抗增殖活性。结果表明，这些化合物对 HL-7702（人肝细胞）和 SKOV-3 的毒性较小，尽管它们的所有活性效力都是中等或较弱的 MDA-MB-231、MCF-7 和 BEL-7404 细胞系。O ^{1 -}炔丙基-N ⁴、N ^{4 -}双(2-氯乙基)-4-氨基苯酚衍生物(化合物III-1~6和VII-1~4 )的抑制活性明显优于N ^{4 -二}炔丙基- N ¹ , N ^{1 -}双（2-氯乙基）-1,4-苯二胺衍生物（化合物IV-1~6和VIII-1~4）针对测试的肿瘤细胞系。并且三个杂交体（化合物III-2、III-6和VII-3）对 BEL-7404 细胞系表现出良好的抑制活性（IC ₅₀分别为 13.5、15.3 和 13.9 μM）。之后，我们对化合物III-6进行KEGG通路分析，结果表明III-6主要通过调节NF-κB通路蛋白的表达来诱导乳腺癌细胞凋亡。