1-Benzyl-2-phenylindole- and 1,2-diphenylindole-based antiestrogens. Estimation of agonist and antagonist activities in transfection assays.

In the 2-phenylindole system, the side chain at the nitrogen atom dominates the endocrine profile both in respect to the reduction of estrogenic action and the increase of antiestrogenic potency. In previous papers we reported on 2-phenylindoles with aliphatic side chains and various functional groups [Biberger, C. and von Angerer, E., J. Steroid Biochem. Molec. Biol., 1996, 58, 31-43 and references therein]. In this study, we incorporated one or two phenyl rings into the side chain in order to lower the flexibility of the side chain. The sulfone group which was used as a polar function was linked to various positions of a benzyl or a phenyl group attached to the indole moiety. The relative binding affinities (RBA) ranged from 1.5 to 8.4% of estradiol. Agonist and antagonist activities were estimated in transfection assays using transiently transfected HeLa cells (cotransfected with the HEG0 vector) and stably transfected MCF-7/2a human breast cancer cells. The reporter plasmid contained the ERE from the Vitellogenin 2A gene, a viral tk promotor and the luciferase gene. Many of the new derivatives showed no or only very low estrogenic activity except for the compound 4e which contained two benzyl elements in the side chain. The antiestrogenic potency was very variable when concentrations 100-fold higher than that of estradiol were applied. The compound with the para-substituted benzyl fragment (4b) proved to be a pure antagonist in the transfection assays. It antagonized the effect of estradiol (10 nM) with an IC50 value of 10(-7) M. It also inhibited strongly the growth of estrogen-sensitive human MCF-7 mammary carcinoma cells (IC50, 3 nM). Its activity was comparable to the one of the corresponding aliphatic 2-phenylindole derivative ZK 164.015. The data from the transcription and proliferation assays suggest that a phenyl ring can be incorporated into the side chain of pure antiestrogens without reducing their potency, provided the aromatic ring is para-substituted and a methylene group between the indole nitrogen and the phenyl group can act as hinge.