Limits...
Development of β-amino-carbonyl compounds as androgen receptor antagonists.

Zhang ZY, Zhu YH, Zhou CH, Liu Q, Lu HL, Ge YJ, Wang MW - Acta Pharmacol. Sin. (2014)

Bottom Line: The therapeutic effects on tumor growth in vivo were observed in male SCID mice bearing LNCaP xenografts.Further structural modifications led to the discovery of a androgen receptor antagonist (compound 6012), which blocked androgen receptor nuclear translocation, androgen-responsive gene expression and androgen-dependent LNCaP cell proliferation.The pharmacological effects of 6012, including AR binding, androgen-induced AR translocation, NH2- and COOH-terminal interaction, growth inhibition of LNCaP cells in vitro and LNCaP xenograft growth in nude mice, were mainly restricted to isomer 6012-4 (1R, 3S).

View Article: PubMed Central - PubMed

Affiliation: The National Center for Drug Screening and the CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences (CAS), Shanghai 201203, China.

ABSTRACT

Aim: Androgen receptor (AR) antagonists have proven to be useful in the early control of prostate cancer. The aim of this study was to identify and characterize a novel β-amino-carbonyl-based androgen receptor antagonist.

Methods: Different isomers of the β-amino-carbonyl compounds were obtained by chiral separation. The bioactivities of the isomers were evaluated by AR nuclear translocation, mammalian two-hybrid, competitive receptor binding and cell proliferation assays. The expression of genes downstream of AR was analyzed with real-time PCR. The therapeutic effects on tumor growth in vivo were observed in male SCID mice bearing LNCaP xenografts.

Results: Compound 21 was previously identified as an AR modulator by the high-throughput screening of a diverse compound library. In the present study, the two isomers of compound 21, termed compounds 21-1 and 21-2, were characterized as partial AR agonists in terms of androgen-induced AR nuclear translocation, prostate-specific antigen expression and cell proliferation. Further structural modifications led to the discovery of a androgen receptor antagonist (compound 6012), which blocked androgen receptor nuclear translocation, androgen-responsive gene expression and androgen-dependent LNCaP cell proliferation. Four stereoisomers of compound 6012 were isolated, and their bioactivities were assessed. The pharmacological effects of 6012, including AR binding, androgen-induced AR translocation, NH2- and COOH-terminal interaction, growth inhibition of LNCaP cells in vitro and LNCaP xenograft growth in nude mice, were mainly restricted to isomer 6012-4 (1R, 3S).

Conclusion: Compound 6012-4 was determined to be a novel androgen receptor antagonist with prostate cancer inhibitory activities comparable to bicalutamide both in vitro and in vivo.

Show MeSH

Related in: MedlinePlus

Biological activities of compounds 21-1 and 21-2 in the androgen receptor signaling pathway. (A) Compounds 21-1 and 21-2 stimulated the nuclear translocation of androgen receptors. (B) Nuclear translocation of androgen receptors in DHT (dihydrotestosterone)-treated LNCaP cells. Compounds were tested together with 10 nmol/L DHT. (C) Effects of compound 21-2 on androgen receptor NC interaction. (D) PSA gene expression. For Figures B, C, and D, the compound concentrations were 100 nmol/L, 1 μmol/L, and 10 μmol/L, from left to right, with or without 10 nmol/L DHT. The data are presented as the mean±SEM of at least three independent experiments. bP<0.05, cP<0.01 compared with 10 nmol/L DHT. fP<0.01 vs DMSO control (t-test). Casodex: bicalutamide.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4814037&req=5

fig1: Biological activities of compounds 21-1 and 21-2 in the androgen receptor signaling pathway. (A) Compounds 21-1 and 21-2 stimulated the nuclear translocation of androgen receptors. (B) Nuclear translocation of androgen receptors in DHT (dihydrotestosterone)-treated LNCaP cells. Compounds were tested together with 10 nmol/L DHT. (C) Effects of compound 21-2 on androgen receptor NC interaction. (D) PSA gene expression. For Figures B, C, and D, the compound concentrations were 100 nmol/L, 1 μmol/L, and 10 μmol/L, from left to right, with or without 10 nmol/L DHT. The data are presented as the mean±SEM of at least three independent experiments. bP<0.05, cP<0.01 compared with 10 nmol/L DHT. fP<0.01 vs DMSO control (t-test). Casodex: bicalutamide.

Mentions: We previously discovered a series of β-amino-carbonyl compounds that function as AR modulators. Compound 21 displayed a high binding affinity and showed AR-modulating bioactivities both in vitro and in vivo10. To explore the underlying molecular mechanism of compound 21's bioactivities, we performed nuclear translocation assays with a high content screening system (Cellomics ArrayScan). As shown in Figure 1A, two isomers of compound 21, (R)-1-(4-chlorophenyl)-3-(furan-2-yl)-3-((4-nitrophenyl)amino)propan-1-one (21-1) and (S)-1-(4-chlorophenyl)-3-(furan-2-yl)-3-[(4-nitrophenyl)amino]propan-1-one (21-2), stimulated AR nuclear translocation, with EC50 values of 14.46 nmol/L and 7.12 nmol/L, respectively. The EC50 value for the control ligand DHT was 0.22 nmol/L (Figure 1A). In addition, compound 21-1 also demonstrated an antagonistic activity in the presence of 10 nmol/L DHT (Figure 1B). These results were subsequently confirmed by image analysis with confocal microscopy in LNCaP cells (Supplemental Figure 1).


Development of β-amino-carbonyl compounds as androgen receptor antagonists.

Zhang ZY, Zhu YH, Zhou CH, Liu Q, Lu HL, Ge YJ, Wang MW - Acta Pharmacol. Sin. (2014)

Biological activities of compounds 21-1 and 21-2 in the androgen receptor signaling pathway. (A) Compounds 21-1 and 21-2 stimulated the nuclear translocation of androgen receptors. (B) Nuclear translocation of androgen receptors in DHT (dihydrotestosterone)-treated LNCaP cells. Compounds were tested together with 10 nmol/L DHT. (C) Effects of compound 21-2 on androgen receptor NC interaction. (D) PSA gene expression. For Figures B, C, and D, the compound concentrations were 100 nmol/L, 1 μmol/L, and 10 μmol/L, from left to right, with or without 10 nmol/L DHT. The data are presented as the mean±SEM of at least three independent experiments. bP<0.05, cP<0.01 compared with 10 nmol/L DHT. fP<0.01 vs DMSO control (t-test). Casodex: bicalutamide.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4814037&req=5

fig1: Biological activities of compounds 21-1 and 21-2 in the androgen receptor signaling pathway. (A) Compounds 21-1 and 21-2 stimulated the nuclear translocation of androgen receptors. (B) Nuclear translocation of androgen receptors in DHT (dihydrotestosterone)-treated LNCaP cells. Compounds were tested together with 10 nmol/L DHT. (C) Effects of compound 21-2 on androgen receptor NC interaction. (D) PSA gene expression. For Figures B, C, and D, the compound concentrations were 100 nmol/L, 1 μmol/L, and 10 μmol/L, from left to right, with or without 10 nmol/L DHT. The data are presented as the mean±SEM of at least three independent experiments. bP<0.05, cP<0.01 compared with 10 nmol/L DHT. fP<0.01 vs DMSO control (t-test). Casodex: bicalutamide.
Mentions: We previously discovered a series of β-amino-carbonyl compounds that function as AR modulators. Compound 21 displayed a high binding affinity and showed AR-modulating bioactivities both in vitro and in vivo10. To explore the underlying molecular mechanism of compound 21's bioactivities, we performed nuclear translocation assays with a high content screening system (Cellomics ArrayScan). As shown in Figure 1A, two isomers of compound 21, (R)-1-(4-chlorophenyl)-3-(furan-2-yl)-3-((4-nitrophenyl)amino)propan-1-one (21-1) and (S)-1-(4-chlorophenyl)-3-(furan-2-yl)-3-[(4-nitrophenyl)amino]propan-1-one (21-2), stimulated AR nuclear translocation, with EC50 values of 14.46 nmol/L and 7.12 nmol/L, respectively. The EC50 value for the control ligand DHT was 0.22 nmol/L (Figure 1A). In addition, compound 21-1 also demonstrated an antagonistic activity in the presence of 10 nmol/L DHT (Figure 1B). These results were subsequently confirmed by image analysis with confocal microscopy in LNCaP cells (Supplemental Figure 1).

Bottom Line: The therapeutic effects on tumor growth in vivo were observed in male SCID mice bearing LNCaP xenografts.Further structural modifications led to the discovery of a androgen receptor antagonist (compound 6012), which blocked androgen receptor nuclear translocation, androgen-responsive gene expression and androgen-dependent LNCaP cell proliferation.The pharmacological effects of 6012, including AR binding, androgen-induced AR translocation, NH2- and COOH-terminal interaction, growth inhibition of LNCaP cells in vitro and LNCaP xenograft growth in nude mice, were mainly restricted to isomer 6012-4 (1R, 3S).

View Article: PubMed Central - PubMed

Affiliation: The National Center for Drug Screening and the CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences (CAS), Shanghai 201203, China.

ABSTRACT

Aim: Androgen receptor (AR) antagonists have proven to be useful in the early control of prostate cancer. The aim of this study was to identify and characterize a novel β-amino-carbonyl-based androgen receptor antagonist.

Methods: Different isomers of the β-amino-carbonyl compounds were obtained by chiral separation. The bioactivities of the isomers were evaluated by AR nuclear translocation, mammalian two-hybrid, competitive receptor binding and cell proliferation assays. The expression of genes downstream of AR was analyzed with real-time PCR. The therapeutic effects on tumor growth in vivo were observed in male SCID mice bearing LNCaP xenografts.

Results: Compound 21 was previously identified as an AR modulator by the high-throughput screening of a diverse compound library. In the present study, the two isomers of compound 21, termed compounds 21-1 and 21-2, were characterized as partial AR agonists in terms of androgen-induced AR nuclear translocation, prostate-specific antigen expression and cell proliferation. Further structural modifications led to the discovery of a androgen receptor antagonist (compound 6012), which blocked androgen receptor nuclear translocation, androgen-responsive gene expression and androgen-dependent LNCaP cell proliferation. Four stereoisomers of compound 6012 were isolated, and their bioactivities were assessed. The pharmacological effects of 6012, including AR binding, androgen-induced AR translocation, NH2- and COOH-terminal interaction, growth inhibition of LNCaP cells in vitro and LNCaP xenograft growth in nude mice, were mainly restricted to isomer 6012-4 (1R, 3S).

Conclusion: Compound 6012-4 was determined to be a novel androgen receptor antagonist with prostate cancer inhibitory activities comparable to bicalutamide both in vitro and in vivo.

Show MeSH
Related in: MedlinePlus