Kym P R, Hummert K L, Nilsson A G, Lubin M, Katzenellenbogen J A
Department of Chemistry, University of Illinois, Urbana 61801, USA.
J Med Chem. 1996 Dec 6;39(25):4897-904. doi: 10.1021/jm960300k.
We have isolated two bisphenolic compounds (4 and 5) that have a marked effect on K+ and Na+ concentrations in human cells from commercial preparations of the pH indicator dye phenol red (phenolsulfonphthalein). We used a bioassay to identify active chromatographic fractions from the lipophilic impurities present in phenol red, and we determined the structure of two active components (4 and 5) by 1H and 13C NMR and mass spectrometry. When added to human fibroblasts in serum-free medium, the bisphenol fluorene derivative 9,9-bis(4'-hydroxyphenyl)-3-hydroxyfluorene (5) produced a rapid loss of K+ and a gain of Na+, at low concentrations, with an EC50 between 30 and 60 ng/mL (80-160 nM). The 2- and 4-hydroxy isomers of the fluorene 5 (i.e., compounds 6 and 7), prepared by synthesis, had similar activity, although compound 6 was somewhat less potent. The bisphenol xanthene derivative 9,9-bis(4'-hydroxyphenyl)xanthene (4) elicited a similar biological response but was less potent than 5-7; it also had a strong effect on cell adhesion, causing release of cells from the plastic substrate at concentrations as low as 2-5 microg/mL (5.5-14 microM). The structures of xanthene (4) and fluorene (5) bisphenols have been confirmed by synthesis from xanthone and hydroxyfluorenone, respectively, by Friedel-Crafts alkylation with phenol. In the latter case, the desired 3-hydroxyfluorene isomer was formed in situ by rearrangement of the 1-hydroxy isomer.
我们从pH指示剂染料酚红（酚磺酞）的商业制剂中分离出了两种双酚类化合物（4和5），它们对人体细胞中的钾离子和钠离子浓度有显著影响。我们使用生物测定法从酚红中存在的亲脂性杂质中鉴定出活性色谱馏分，并通过¹H和¹³C核磁共振以及质谱法确定了两种活性成分（4和5）的结构。当将双酚芴衍生物9,9-双（4'-羟基苯基）-3-羟基芴（5）添加到无血清培养基中的人成纤维细胞时，在低浓度下会导致钾离子迅速流失和钠离子增加，其半数有效浓度（EC50）在30至60 ng/mL（80 - 160 nM）之间。通过合成制备的芴5的2-和4-羟基异构体（即化合物6和7）具有相似的活性，尽管化合物6的效力稍低。双酚呫吨衍生物9,9-双（4'-羟基苯基）呫吨（4）引发了类似的生物学反应，但效力低于5 - 7；它对细胞黏附也有很强的影响，在低至2 - 5 μg/mL（5.5 - 14 μM）的浓度下就能使细胞从塑料基质上脱落。呫吨（4）和芴（5）双酚的结构已分别通过用苯酚进行傅克烷基化反应，从呫吨酮和羟基芴酮合成得到证实。在后一种情况下，所需的3-羟基芴异构体通过1-羟基异构体的重排原位形成。
