Design and Synthesis of 4‑(4-Benzoylaminophenoxy)phenol Derivatives As Androgen Receptor Antagonists

We report the design and synthesis of novel 4-(4-benzoylaminophenoxy)­phenol derivatives that bind to the androgen receptor (AR) ligand-binding domain and exhibit potent androgen-antagonistic activity. Compound <b>22</b> is one of the most potent of these derivatives, inhibiting the dihydrotestosterone-promoted growth of SC-3 cell line bearing wild-type AR (IC₅₀ 0.75 μM), LNCaP cell line bearing T877A-mutated AR (IC₅₀ 0.043 μM), and 22Rv1 cell line bearing H874Y-mutated AR (IC₅₀ 0.22 μM). Structure–activity relationship studies confirmed that the pharmacophore of these novel AR antagonists is distinct from the nitro- or cyano-substituted anilide substructure of other nonsteroidal AR antagonists. This novel pharmacophore is expected to provide a basis for designing new antiprostate cancer agents

Design and Synthesis of 4‑(4-Benzoylaminophenoxy)phenol Derivatives As Androgen Receptor Antagonists

We report the design and synthesis of novel 4-(4-benzoylaminophenoxy)­phenol derivatives that bind to the androgen receptor (AR) ligand-binding domain and exhibit potent androgen-antagonistic activity. Compound <b>22</b> is one of the most potent of these derivatives, inhibiting the dihydrotestosterone-promoted growth of SC-3 cell line bearing wild-type AR (IC₅₀ 0.75 μM), LNCaP cell line bearing T877A-mutated AR (IC₅₀ 0.043 μM), and 22Rv1 cell line bearing H874Y-mutated AR (IC₅₀ 0.22 μM). Structure–activity relationship studies confirmed that the pharmacophore of these novel AR antagonists is distinct from the nitro- or cyano-substituted anilide substructure of other nonsteroidal AR antagonists. This novel pharmacophore is expected to provide a basis for designing new antiprostate cancer agents

Novel Nonsteroidal Progesterone Receptor (PR) Antagonists with a Phenanthridinone Skeleton

The progesterone receptor (PR) plays an important role in various physiological systems, including female reproduction and the central nervous system, and PR antagonists are thought to be effective not only as contraceptive agents and abortifacients but also in the treatment of various diseases, including hormone-dependent cancers and endometriosis. Here, we identified phenanthridin-6­(5<i>H</i>)-one derivatives as a new class of PR antagonists and investigated their structure–activity relationships. Among the synthesized compounds, <b>37</b>, <b>40</b>, and <b>46</b> exhibited very potent PR antagonistic activity with high selectivity for PR over other nuclear receptors. These compounds are structurally distinct from other nonsteroidal PR antagonists, including cyanoaryl derivatives, and should be useful for further studies of the clinical utility of PR antagonists

Development of <i>N</i>‑(4-Phenoxyphenyl)benzenesulfonamide Derivatives as Novel Nonsteroidal Progesterone Receptor Antagonists

We report here development of <i>N</i>-(4-phenoxyphenyl)­benzenesulfonamide derivatives as a novel class of nonsteroidal progesterone receptor (PR) antagonists. PR plays key roles in various physiological systems, including the female reproductive system, and PR antagonists are candidates for clinical treatment of multiple diseases, including uterine leiomyoma, endometriosis, breast cancer, and some psychiatric disorders. We found that the benzenesulfonanilide skeleton functions as a novel scaffold for PR antagonists, and we adopted 3-chlorobenzenesulfonyl derivative <b>20a</b> as a lead compound for structural development. Among the synthesized compounds, 3-trifluoromethyl derivative <b>32</b> exhibited the most potent PR-antagonistic activity, with high binding affinity for PR and selectivity over androgen receptor (AR). It is structurally distinct from other nonsteroidal PR antagonists, including cyanopyrrole derivatives, and further modification is expected to afford novel selective PR modulators

A Modified Murine Embryonic Stem Cell Test for Evaluating the Teratogenic Effects of Drugs on Early Embryogenesis

<div><p>Mammalian fetal development is easily disrupted by exogenous agents, making it essential to test new drug candidates for embryotoxicity and teratogenicity. To standardize the testing of drugs that might be used to treat pregnant women, the U.S. Food and Drug Administration (FDA) formulated special grade categories, labeled A, B, C, D and X, that define the level of risk associated with the use of a specific drug during pregnancy. Drugs in categories (Cat.) D and X are those with embryotoxic and/or teratogenic effects on humans and animals. However, which stages of pregnancy are affected by these agents and their molecular mechanisms are unknown. We describe here an embryonic stem cell test (EST) that classifies FDA pregnancy Cat.D and Cat.X drugs into 4 classes based on their differing effects on primitive streak formation. We show that ~84% of Cat.D and Cat.X drugs target this period of embryogenesis. Our results demonstrate that our modified EST can identify how a drug affects early embryogenesis, when it acts, and its molecular mechanism. Our test may thus be a useful addition to the drug safety testing armamentarium.</p></div

Effects of drug classes on EB gene expression patterns.

<p>(A-C) EBs were treated with the indicated class-representative drugs for days 3–6 of EB culture (>30 EB cultures/group). Samples were collected on days 4–6. DMSO, control; Nandrolone, Class I; acitretin, Class II; trimetrexate, Class III; vincristine, Class IV; SB203580, p38MAPK inhibitor. (A) Quantitative RT-PCR analysis of Wnt3 mRNA on day 6 determined as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0145286#pone.0145286.g001" target="_blank">Fig 1A</a>. **, P<0.01. (B) <i>In situ</i> hybridization to detect Brachyury T on day 6 in EB cultures treated as indicated for days 3–6. Results are representative of 3 independent experiments (15 EBs/group). (C) Quantitative RT-PCR analysis of the indicated mRNAs on the indicated days determined as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0145286#pone.0145286.g001" target="_blank">Fig 1B</a>. (D) Quantitative RT-PCR analysis of Bmp2 mRNA in EB cultures that were treated with DMSO (control) or nandrolone on days 3–6. Data were analyzed as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0145286#pone.0145286.g001" target="_blank">Fig 1A</a>. NS, not significant. (E) Top: Illustration of mouse embryonic and germ layer development at the indicated embryonic days (E). Epi, epiblast; PS, primitive streak. Bottom: Illustration of days (D) of mouse EB culture indicating the percentage of each drug class acting at a particular stage of culture.</p

Teratogenicity of benzodiazepines.

<p>(A) Quantitative RT-PCR determination of mRNA levels of the indicated genes in EB cultures that were treated for days 3–6 with DMSO (control) or the indicated drugs (>30 EBs/group) and evaluated on day 6. Wnt3 and Brachyury T, primitive streak markers; Sox2, ectoderm marker; BMP2, mesoderm marker; GATA6, endoderm marker; Nestin, neuroectoderm marker. Data were analyzed as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0145286#pone.0145286.g001" target="_blank">Fig 1A</a>. *, P<0.05. (B) RT-PCR analysis of expression of the indicated GABA receptor subunits in untreated EBs on the indicated days of culture. Data are representative of 3 independent experiments (>30 EBs/group). (C) <i>In situ</i> hybridization to detect Brachyury T and Sox2 in EBs on day 6 after GABA or alprazolam treatment at the indicated concentrations (15 EBs/group). (D) <i>In situ</i> hybridization to detect Brachyury T and Sox2 in EBs that were treated for 6 days with alprazolam and/or GABA as indicated (20 EBs/group). (E) A proposed model of how BZ may affect primitive streak formation. Treatment with GABA or BZ alone induces only weak signals that cannot inhibit primitive streak formation. However, a combination of BZ plus GABA works synergistically and may induce a strong signal that suppresses primitive streak formation, impairing embryogenesis.</p

<i>In vivo</i> effects of retinoic acid on early mouse embryogenesis.

<p>(A) Scheme illustrating the experimental timeline during which pregnant mice received intravenous injection of 25 mg/kg RA at E5.5, E6.5, E7.5, E8.5 or E9.5. Mice were sacrificed on E10.5, E13.5 or E14.5 and macroscopic effects on uteri and embryo development were determined. (B) Representative images of morphological changes observed in embryos in the uteri of mice that were treated with RA (+) or not (-) at the indicated stages and examined at E10.5. ND, not detected. (C) Representative images of morphological changes observed in embryos in the uteri of mice that were treated with RA at E8.5 and examined at E13.5, or treated at E9.5 and examined at E14.5.</p

Classification of FDA pregnancy category D and X drugs into four classes of potential teratogens.

<p>(A) Scheme illustrating the growth and phenotypes of representative EBs treated with the indicated class-representative drugs for the indicated times of EB development. “Beating”, cardiomyocytes were present in the EB. “Neurite”, neuronal differentiation was observed. EBs treated with Class I drugs displayed beating and no neurites. Class II drug-treated EBs showed neurites with or without foci of beating cardiomyocytes. Class III drug-treated EBs showed neither beating nor neurites but the EBs survived. Class IV drugs resulted in dead EBs. -, EBs attached to the plate but no beating observed. *, data could not be obtained because dead EBs did not attach to the plate. Images of DMSO-treated control EBs at the same stages are also shown. (B) Pie charts of distribution of the indicated drug types among Classes I–IV. (C) Pie charts of numbers of total, Cat.D and Cat.X drugs in Classes I–IV.</p

Crystal Engineering of <i>N</i>,<i>N</i>′‑Diphenylurea Compounds Featuring Phenyl–Perfluorophenyl Interaction

Here, aiming to adopt the phenyl–perfluorophenyl interaction to regulate molecular alignment and arrangement for crystal engineering, we examined and compared in detail the crystal structures of <i>N</i>,<i>N</i>′-diphenylurea compounds <b>1</b>–<b>6</b>. We found that phenyl–perfluorophenyl interaction greatly influenced the intermolecular arrangement in the crystal, and we were able to prepare a cocrystal of <b>1</b> and <b>2</b>, in which the molecules were alternately arranged under the control of the phenyl–perfluorophenyl interaction. This arrangement was driven by the asymmetric geometry of the hydrogen bonds in the cocrystal (<b>1·2</b>), in which <b>2</b>, bearing two perfluorophenyl groups, worked as a better hydrogen bond donor. In contrast, NH connected to the phenyl group in <b>3</b> proved to be a better hydrogen bond donor due to the intramolecular resonance effect. <i>N</i>,<i>N</i>′-Dimethylated derivatives, <b>4</b>–<b>6</b>, existed in <i>cis</i>-<i>cis</i> form in the crystal. Antiparallel carbonyl–carbonyl arrangements were observed in <b>4</b> and <b>6</b>, while an unexpected carbonyl–perfluorophenyl interaction was observed in the crystal of <b>5</b>. These findings will be helpful in the design of diphenylurea-based functional molecules, especially for solid-state application