Synthesis and biological assessment of chalcone and pyrazoline derivatives as novel inhibitor for ELF3-MED23 interaction
- College of Pharmacy and Graduate School of Pharmaceutical Sciences, Ewha Womans University, Republic of Korea
- College of Pharmacy, CHA University, Republic of Korea
Figures
Figure 1
Structure of proposed compounds.
Figure 2
Structures of the prepared compounds.
Figure 3
Identification of compound 10 as potent ELF3-MED23 PPI inhibitor.
(A) Synthesized compounds were screened to evaluate their inhibitory activity against ELF3–MED23 PPI. All compounds were used at 10 μM for 12 h (n=3, mean ± S.D.). (B) Cell viability was also …
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Figure 3—source data 1
Raw unedited gels for Figure 3C.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig3-data1-v1.zip
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Figure 3—source data 2
Uncropped and labelled gels for Figure 3C.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig3-data2-v1.zip
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Figure 3—source data 3
Raw unedited gels for Figure 3E.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig3-data3-v1.zip
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Figure 3—source data 4
Uncropped and labelled gels for Figure 3E.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig3-data4-v1.zip
Figure 4
Qualitative SAR of prepared compounds.
Activity cliff summary for the prepared compound series were plotted regarding the contributions of electrostatic (A) and hydrophobic (B) fields. The field point pattern was depicted as spheres …
Figure 5
Compound 10 as a transcriptional regulator of HER2 by inhibiting ELF3-MED23 PPI.
(A) Titration curve of MED23391-582 protein FITC-labeled ELF3129-145 peptide. Binding of the MED23391-582 protein and ELF3-FITC peptide (17 a.a.) was validated via cell-free FP assay. Kd value was …
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Figure 5—source data 1
Raw unedited gels for Figure 5C.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig5-data1-v1.zip
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Figure 5—source data 2
Uncropped and labeled gels for Figure 5C.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig5-data2-v1.zip
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Figure 5—source data 3
Raw unedited gels for Figure 5F.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig5-data3-v1.zip
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Figure 5—source data 4
Uncropped and labelled gels for Figure 5F.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig5-data4-v1.zip
Figure 6
Compound 10 as a potent anticancer agent for HER2-positive gastric cancer cells.
(A) Apoptosis induced by compound 10 in parental NCI-N87 was assessed by treatment with the compound in a dose-dependent manner (24 hr treatment at indicated concentrations, n=3, mean ± S.D., ANOVA, …
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Figure 6—source data 1
Raw unedited gels for Figure 6B.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig6-data1-v1.zip
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Figure 6—source data 2
Uncropped and labelled gels for Figure 6B.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig6-data2-v1.zip
Figure 7
Compound 10 as a novel strategy to overcome trastuzumab resistance.
(A) Trastuzumab (10 μg/ml, 24 hr)-induced growth inhibitory effects were assessed against parental and TR NCI-N87 cell lines (n=3, mean ± S.D., Student’s t-test, ***p<0.001 vs. parent). (B) …
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Figure 7—source data 1
Raw unedited gels for Figure 7B.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig7-data1-v1.zip
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Figure 7—source data 2
Uncropped and labelled gels for Figure 7B.
- https://cdn.elifesciences.org/articles/97051/elife-97051-fig7-data2-v1.zip
Scheme 1
General synthesis methods for the target compounds.
Author response image 1
HER2 downregulatory effect of compound 10 in HER2-positive breast cancer cell lines, AU565 and BT474.
Author response image 2
Docking analysis of compound 10.
Author response image 3
Impact of methoxy group introduction on TPSA (total polar surface area) of each molecule.
TPSA of each molecule containing chalcone structure were calculated using the Molinspiration webserver.
Author response image 4
HER2 downregulatory effect of compound 10 in HER2-positive gastric cancer cell line, SNU216.
(A) Expression levels of HER2 and ELF3 in various gastric cancer cell lines. (B) HER2 downregulation in the SNU216 cell line following treatment with compound 10.
Tables
Table 1
Summary of the antibody lists used in this study.
| Antibody | Vendor | Catalogue number | Application | Dilution |
|---|---|---|---|---|
| AKT | Cell signaling | #9272 | WB | 1:2000 |
| c-PARP | Cell signaling | #9541 | WB | 1:2000 |
| ELF3 | Thermo Fisher | PA5-21293 | WB | 1:2000 |
| FLAG | MBL | M185-3L | WB | 1:1000 |
| GAPDH | MBL | M171-3 | WB | 1:10000 |
| GST | MBL | M209-3 | WB | 1:1000 |
| HER2 | Thermo Fisher | MA5-13105 | WB | 1:1000 |
| IHC | 1:100 | |||
| Ki67 | Dako | M7240 | IHC | 1:200 |
| MAPK | Cell signaling | #9102 | WB | 1:2000 |
| MED23 | Novus | NB200-338 | WB | 1:2000 |
| p-AKT (S473) | Santa cruz | sc-7985 | WB | 1:2000 |
| p-MAPK (T202/Y204) | Cell signaling | #9101 | WB | 1:2000 |
| Cleaved caspase-3 | Cell signaling | #9661 | WB | 1:1000 |
Table 2
Information of utilized qRT-PCR primers.
| Gene | Sequence | |
|---|---|---|
| Actin | Forward | 5' AGCCATGTACGTAGCCATCC 3' |
| Reverse | 5' CTCTCAGCTGTGGTGGTGAA 3' | |
| ERBB2 | Forward | 5' GGTGGTCTTTGGGATCCTCA 3' |
| Reverse | 5' ACCTTCACCTTCCTCAGCTC 3' | |
| ELF3 | Forward | 5' GTGATGCTGAGCTTGGGATG 3' |
| Reverse | 5' TTAGGTTAGAAGCGCCCACA 3' | |
| GAPDH | Forward | 5' GAGTCAACGGATTTGGTCGT 3' |
| Reverse | 5' GACAAGCTTCCCGTTCTCAG 3' |
Additional files
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MDAR checklist
- https://cdn.elifesciences.org/articles/97051/elife-97051-mdarchecklist1-v1.docx
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Supplementary file 1
1H NMR and 13C NMR Spectra of compounds 1–26.
- https://cdn.elifesciences.org/articles/97051/elife-97051-supp1-v1.xlsx
