Korach K S, Metzler M, McLachlan J A
Proc Natl Acad Sci U S A. 1978 Jan;75(1):468-71. doi: 10.1073/pnas.75.1.468.
The diethylstilbestrol (DES) metabolite (beta-dienestrol), which had been identified in mouse, rat, monkey, and human urine, and two proposed metabolic intermediates (diethylstilbestrol alpha,alpha'-epoxide and alpha,alpha'-dihydroxy DES) were synthesized and their estrogenic activities determined. In addition, three DES analogs, alpha-dienestrol, DES-dihydroxy diethyl phenanthrene (DES-phenanthrene), and 1-(alpha-ethyl, 4alpha-hydroxyphenyl)indanyl-5-ol (indanyl-DES), were studied. Estrogenic activities of the compounds in vivo were determined by the immature mouse uterine weight bioassay; in vitro, their estradiol receptor binding activity (competitive equilibrium binding, sucrose gradient analysis, and association rate inhibition assays) was determined. Results of the mouse uterine weight bioassay gave the following order of estrogenicity: DES > alpha-dienestrol >/= DES-epoxide > indanyl-DES > dihydroxy DES > beta-dienestrol > DES-phenanthrene. Results of competitive equilibrium binding analyses of these compounds with estradiol-17beta for the mouse uterine cytosol receptor followed the same order seen for the bioassay, except for indanyl-DES. DES, indanyl-DES, and alpha-dienestrol had the greatest affinities (K(a) values approximately 0.5-19.1 x 10(10) M(-1)), while DES-phenanthrene had the lowest (K(a) = 3.5 x 10(7) M(-1) +/- 1.2). Sucrose gradient analysis of the above competition preparations illustrated the displacement of [(3)H]estradiol from the receptor peak. This displacement was receptor specific and concentration dependent and correlated with the equilibrium binding concentrations. In addition, the most hormonally active substances demonstrated the greatest rate inhibition in the estradiol cytosol receptor association rate reaction (V(0)). The rank order of estrogenicity of the compounds determined in this study should be useful in evaluating alternative metabolic pathways of DES as well as distinguishing biologically active metabolites from relatively inactive ones.
已在小鼠、大鼠、猴和人类尿液中鉴定出的己烯雌酚（DES）代谢物（β-二烯雌酚），以及两种推测的代谢中间体（己烯雌酚α,α'-环氧化物和α,α'-二羟基DES）被合成出来，并测定了它们的雌激素活性。此外，还研究了三种DES类似物，即α-二烯雌酚、DES-二羟基二乙基菲（DES-菲）和1-(α-乙基, 4α-羟基苯基)茚满-5-醇（茚满-DES）。通过未成熟小鼠子宫重量生物测定法测定了这些化合物在体内的雌激素活性；在体外，测定了它们的雌二醇受体结合活性（竞争性平衡结合、蔗糖梯度分析和结合速率抑制试验）。小鼠子宫重量生物测定法的结果给出了以下雌激素活性顺序：DES＞α-二烯雌酚≥DES-环氧化物＞茚满-DES＞二羟基DES＞β-二烯雌酚＞DES-菲。这些化合物与小鼠子宫胞质溶胶受体的雌二醇-17β进行竞争性平衡结合分析的结果，除茚满-DES外，遵循与生物测定法相同的顺序。DES、茚满-DES和α-二烯雌酚具有最大亲和力（K(a)值约为0.5 - 19.1×10(10) M(-1)），而DES-菲的亲和力最低（K(a) = 3.5×10(7) M(-1)±1.2）。上述竞争制剂的蔗糖梯度分析表明[(3)H]雌二醇从受体峰上被置换下来。这种置换是受体特异性的且浓度依赖性的，并且与平衡结合浓度相关。此外，激素活性最强的物质在雌二醇胞质溶胶受体结合速率反应（V(0)）中表现出最大的速率抑制。本研究中确定的化合物雌激素活性顺序，对于评估DES的替代代谢途径以及区分生物活性代谢物和相对无活性的代谢物应是有用的。
