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Discovery of novel aromatase inhibitors using a homogeneous time-resolved fluorescence assay.

Ji JZ, Lao KJ, Hu J, Pang T, Jiang ZZ, Yuan HL, Miao JS, Chen X, Ning SS, Xiang H, Guo YM, Yan M, Zhang LY - Acta Pharmacol. Sin. (2014)

Bottom Line: Among the 7000 compounds, 4 hits (XHN22, XHN26, XHN27 and triptoquinone A) were found to inhibit aromatase with IC50 values of 1.60±0.07, 2.76±0.24, 0.81±0.08 and 45.8±11.3 μmol /L, respectively.Moreover, the most potent hit XHN27 at 10 and 50 μmol/L inhibited the proliferation of T47D cells by 45.3% and 35.2%, respectively.The docking study revealed that XHN27 docked within the active site of aromatase and might form a hydrogen bond and had a π-cation interaction with amino acid residues of the protein.

View Article: PubMed Central - PubMed

Affiliation: Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing 210009, China.

ABSTRACT

Aim: Aromatase is an important target for drugs to treat hormone-dependent diseases, including breast cancer. The aim of this study was to develop a homogeneous time-resolved fluorescence (HTRF) aromatase assay suitable for high-throughput screening (HTS).

Methods: A 384-well aromatase HTRF assay was established, and used to screen about 7000 compounds from a compound library. Anti-proliferation activity of the hit was evaluated using alamarBlue(R) assay in a hormone-dependent breast cancer cell line T47D. Molecular docking was conducted to elucidate the binding mode of the hit using the Discovery Studio program.

Results: The Z' value and signal to background (S/B) ratio were 0.74 and 5.4, respectively. Among the 7000 compounds, 4 hits (XHN22, XHN26, XHN27 and triptoquinone A) were found to inhibit aromatase with IC50 values of 1.60±0.07, 2.76±0.24, 0.81±0.08 and 45.8±11.3 μmol /L, respectively. The hits XHN22, XHN26 and XHN27 shared the same chemical scaffold of 4-imidazolyl quinoline. Moreover, the most potent hit XHN27 at 10 and 50 μmol/L inhibited the proliferation of T47D cells by 45.3% and 35.2%, respectively. The docking study revealed that XHN27 docked within the active site of aromatase and might form a hydrogen bond and had a π-cation interaction with amino acid residues of the protein.

Conclusion: XHN27, an imidazolyl quinoline derivative of flavonoid, is a potent aromatase inhibitor with anti-proliferation activity against breast cancer in vitro. The established assay can be used in HTS for discovering novel aromatase inhibitor.

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The skeleton of the newly discovered aromatase inhibitor.
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fig5: The skeleton of the newly discovered aromatase inhibitor.

Mentions: Compounds were tested at a single concentration of 0.1 mmol/L in duplicate. The primary screening was performed with 15 nmol/L aromatase, 30 nmol/L testosterone, and 600 nmol/L NADPH and resulted in 5 hits. The secondary screening resulted in 4 confirmed hits, XHN22, XHN26, XHN27, and triptoquinone A (TQA) (Figure 4A–4D), representing a hit rate of 0.067%. Three of the 4 compounds (XHN22, XHN26 and XHN27), derivatives of flavone, share the general chemical skeleton of imidazolyl quinoline (Figure 5) and had an IC50 against aromatase of 1.60±0.07 μmol/L, 2.76±0.24 μmol/L, and 0.81±0.08 μmol/L, respectively (Figure 4A–4C). The IC50 of biochanin A (BCA) was 30.0±3.3 μmol/L (Figure 3B), indicating that BCA possesses a relatively weak activity against aromatase compared with the imidazolyl quinoline derivatives discovered in this study. The other confirmed hit, TQA (Figure 4D), extracted from Tripterygium wilfordii, was less potent compared with XHN27 (Figure 4C), with an IC50 against aromatase of 45.8±11.3 μmol/L.


Discovery of novel aromatase inhibitors using a homogeneous time-resolved fluorescence assay.

Ji JZ, Lao KJ, Hu J, Pang T, Jiang ZZ, Yuan HL, Miao JS, Chen X, Ning SS, Xiang H, Guo YM, Yan M, Zhang LY - Acta Pharmacol. Sin. (2014)

The skeleton of the newly discovered aromatase inhibitor.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4125720&req=5

fig5: The skeleton of the newly discovered aromatase inhibitor.
Mentions: Compounds were tested at a single concentration of 0.1 mmol/L in duplicate. The primary screening was performed with 15 nmol/L aromatase, 30 nmol/L testosterone, and 600 nmol/L NADPH and resulted in 5 hits. The secondary screening resulted in 4 confirmed hits, XHN22, XHN26, XHN27, and triptoquinone A (TQA) (Figure 4A–4D), representing a hit rate of 0.067%. Three of the 4 compounds (XHN22, XHN26 and XHN27), derivatives of flavone, share the general chemical skeleton of imidazolyl quinoline (Figure 5) and had an IC50 against aromatase of 1.60±0.07 μmol/L, 2.76±0.24 μmol/L, and 0.81±0.08 μmol/L, respectively (Figure 4A–4C). The IC50 of biochanin A (BCA) was 30.0±3.3 μmol/L (Figure 3B), indicating that BCA possesses a relatively weak activity against aromatase compared with the imidazolyl quinoline derivatives discovered in this study. The other confirmed hit, TQA (Figure 4D), extracted from Tripterygium wilfordii, was less potent compared with XHN27 (Figure 4C), with an IC50 against aromatase of 45.8±11.3 μmol/L.

Bottom Line: Among the 7000 compounds, 4 hits (XHN22, XHN26, XHN27 and triptoquinone A) were found to inhibit aromatase with IC50 values of 1.60±0.07, 2.76±0.24, 0.81±0.08 and 45.8±11.3 μmol /L, respectively.Moreover, the most potent hit XHN27 at 10 and 50 μmol/L inhibited the proliferation of T47D cells by 45.3% and 35.2%, respectively.The docking study revealed that XHN27 docked within the active site of aromatase and might form a hydrogen bond and had a π-cation interaction with amino acid residues of the protein.

View Article: PubMed Central - PubMed

Affiliation: Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing 210009, China.

ABSTRACT

Aim: Aromatase is an important target for drugs to treat hormone-dependent diseases, including breast cancer. The aim of this study was to develop a homogeneous time-resolved fluorescence (HTRF) aromatase assay suitable for high-throughput screening (HTS).

Methods: A 384-well aromatase HTRF assay was established, and used to screen about 7000 compounds from a compound library. Anti-proliferation activity of the hit was evaluated using alamarBlue(R) assay in a hormone-dependent breast cancer cell line T47D. Molecular docking was conducted to elucidate the binding mode of the hit using the Discovery Studio program.

Results: The Z' value and signal to background (S/B) ratio were 0.74 and 5.4, respectively. Among the 7000 compounds, 4 hits (XHN22, XHN26, XHN27 and triptoquinone A) were found to inhibit aromatase with IC50 values of 1.60±0.07, 2.76±0.24, 0.81±0.08 and 45.8±11.3 μmol /L, respectively. The hits XHN22, XHN26 and XHN27 shared the same chemical scaffold of 4-imidazolyl quinoline. Moreover, the most potent hit XHN27 at 10 and 50 μmol/L inhibited the proliferation of T47D cells by 45.3% and 35.2%, respectively. The docking study revealed that XHN27 docked within the active site of aromatase and might form a hydrogen bond and had a π-cation interaction with amino acid residues of the protein.

Conclusion: XHN27, an imidazolyl quinoline derivative of flavonoid, is a potent aromatase inhibitor with anti-proliferation activity against breast cancer in vitro. The established assay can be used in HTS for discovering novel aromatase inhibitor.

Show MeSH
Related in: MedlinePlus