Selective agonism of the estrogen receptor (ER) subtypes, ERα and ERβ, has historically been difficult to achieve due to the high degree of ligand-binding domain structural similarity. Multiple efforts have focused on the use of classical organic scaffolds to model 17β-estradiol geometry in the design of ERβ selective agonists, with several proceeding to various stages of clinical development. Carborane scaffolds offer many unique advantages including the potential for novel ligand/receptor interactions but remain relatively unexplored. We synthesized a series of para-carborane estrogen receptor agonists revealing an ERβ selective structure-activity relationship. We report ERβ agonists with low nanomolar potency, greater than 200-fold selectivity for ERβ over ERα, limited off-target activity against other nuclear receptors, and only sparse CYP450 inhibition at very high micromolar concentrations. The pharmacological properties of our para-carborane ERβ selective agonists measure favorably against clinically developed ERβ agonists and support further evaluation of carborane-based selective estrogen receptor modulators.

• MeSH
• beta receptor estrogenů agonisté   MeSH
• estrogeny chemická syntéza   chemie   farmakologie   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• molekulární struktura MeSH
• sloučeniny boru chemická syntéza   chemie   farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

• Publikační typ
• abstrakty MeSH

Steroid hormone receptors represent a major target in drug discovery. As ligand inducible transcription factors, their activity can be modulated by small lipophilic molecules. Here we describe two panels of potent and selective luciferase reporter cell lines based on cells with low endogenous steroid receptor activity (U2OS). The panels contain reporter cell lines for estrogen receptors α and β, androgen, glucocorticoid, mineralocorticoid, and progesterone receptors. In the first panel, the activation of either synthetic, steroid response elements containing promoter or viral promoter is mediated by full-length steroid receptors. The second panel is based on the expression of the chimeric receptor, which was created by the replacement of the N-terminal part of the molecule by Gal4 DBD and that binds to multiple UAS sites in the reporter promoter. Both panels were extensively characterized by profiling 28 ligands in dose response manner in agonist and antagonist mode. We have analyzed and compared the responses to tested ligands from both panels and concluded that in general both systems generated similar qualitative response in terms of potency, efficacy, partial agonism/antagonism, mixed agonistic/antagonistic profiles and the rank of potencies was well conserved between both panels. However, we have also identified some artifacts introduced by the Gal4/LBD reporter assays in contrast to their full-length receptor reporter counterparts. Keeping in mind the advantages and drawbacks of each reporter format, these cell lines represent powerful and selective tools for profiling large compound libraries (HTS) and for detailed study of mechanisms by which compounds exert their biological effects.

• MeSH
• aktivace transkripce účinky léků   MeSH
• genetické inženýrství metody   MeSH
• knihovny malých molekul analýza   MeSH
• lidé MeSH
• luciferasy genetika   metabolismus   MeSH
• nádorové buněčné linie MeSH
• objevování léků metody   MeSH
• plazmidy genetika   MeSH
• promotorové oblasti (genetika) účinky léků   MeSH
• rekombinantní proteiny genetika   metabolismus   MeSH
• reportérové geny MeSH
• rychlé screeningové testy metody   MeSH
• sekvenční delece MeSH
• steroidní receptory agonisté   antagonisté a inhibitory   genetika   metabolismus   MeSH
• steroidy farmakologie   MeSH
• transfekce MeSH
• transkripční faktory genetika   metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

To identify novel estrogen receptor ligands a series of substituted 17alpha-arylestradiols were synthesized using the catalytic [2 + 2 + 2]cyclotrimerization of 17alpha-ethynylestradiol with various 1,7-diynes in the presence of Wilkinson's catalysts [Rh(PPh(3))(3)Cl]. The compounds were subjected to competitive binding assays, cell-based luciferase reporter assays, and proliferation assays. These experiments confirmed their estrogenic character and revealed some interesting properties like mixed partial/full agonism for ERalpha/ERbeta and different selectivity as a result of differing potencies for either ER.

• MeSH
• aktivace transkripce MeSH
• alfa receptor estrogenů agonisté   genetika   MeSH
• alkyny chemie   MeSH
• beta receptor estrogenů agonisté   genetika   MeSH
• buněčné linie MeSH
• cyklizace MeSH
• estradiol analogy a deriváty   chemická syntéza   farmakologie   MeSH
• fluorescenční polarizace MeSH
• kompetitivní vazba MeSH
• lidé MeSH
• ligandy MeSH
• luciferasy genetika   MeSH
• nádorové buněčné linie MeSH
• parciální agonismus léků MeSH
• proliferace buněk účinky léků   MeSH
• reportérové geny MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH