Abstract
Currently used antiestrogenic drugs against hormone‐dependent breast cancer, and estrogenic drugs used in treatment of osteoporosis, are associated with risk factors. Therefore, there is a strong need to develop selective estrogen receptor modulators with better tissue selectivity. In a recent study (Peptides, 2002, Vol. 3, 573–580), we used a monoclonal antibody to estradiol (mAb‐E2) to screen a phage‐display peptide library. We identified a 15‐mer peptide (peptide H5) that recognizes mAb‐E2 (IC~50~ 1 μM) and estrogen receptor (ER)α (IC~50~ 500 μM) but not ERβ, and displays estrogen‐like activity in vitro and in vivo. In this study, we designed and prepared peptides based on peptide H5, which possess improved estrogenic activity, by evaluating their binding to mAb‐E2 and to ERs. Initially, we determined the minimal binding sequence of peptide H5 capable of binding mAb‐E2 and ER. Subsequently, systematic single‐residue replacements of the minimal sequence, followed by multiple‐residue replacements, yielded hexa‐ and heptapeptides with increased affinities to mAb‐E2 and to ER. The most promising peptides, VSWFFE (EMP‐1) and VSWFFED (EMP‐2) (EMP: estrogen‐mimetic peptide), bind mAb‐E2 with high affinity (IC~50~ of 6 and 30 nM, respectively), recognize ERs with increased affinity (IC~50~ of 100 μM for ERα, and 100–250 μM for ERβ), and possess estrogenic activity in vivo. The short peptides described in this study may be used as potential lead compounds for developing new ER ligands. © 2004 Wiley Periodicals, Inc. Biopolymers (Pept Sci), 2004