Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancer
Three series of compounds were prioritized from a high content screening campaign that identified molecules that blocked dihydrotestosterone (DHT) induced formation of Androgen Receptor (AR) protein-protein interactions (PPIs) with the Transcriptional Intermediary Factor 2 (TIF2) coactivator and als...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-10-01
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| Series: | SLAS Discovery |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472555223000539 |
| _version_ | 1797647527790510080 |
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| author | Ashley T. Fancher Yun Hua David A. Close Wei Xu Lee A. McDermott Christopher J. Strock Ulises Santiago Carlos J. Camacho Paul A. Johnston |
| author_facet | Ashley T. Fancher Yun Hua David A. Close Wei Xu Lee A. McDermott Christopher J. Strock Ulises Santiago Carlos J. Camacho Paul A. Johnston |
| author_sort | Ashley T. Fancher |
| collection | DOAJ |
| description | Three series of compounds were prioritized from a high content screening campaign that identified molecules that blocked dihydrotestosterone (DHT) induced formation of Androgen Receptor (AR) protein-protein interactions (PPIs) with the Transcriptional Intermediary Factor 2 (TIF2) coactivator and also disrupted preformed AR-TIF2 PPI complexes; the hydrobenzo-oxazepins (S1), thiadiazol-5-piperidine-carboxamides (S2), and phenyl-methyl-indoles (S3). Compounds from these series inhibited AR PPIs with TIF2 and SRC-1, another p160 coactivator, in mammalian 2-hybrid assays and blocked transcriptional activation in reporter assays driven by full length AR or AR-V7 splice variants. Compounds inhibited the growth of five prostate cancer cell lines, with many exhibiting differential cytotoxicity towards AR positive cell lines. Representative compounds from the 3 series substantially reduced both endogenous and DHT-enhanced expression and secretion of the prostate specific antigen (PSA) cancer biomarker in the C4–2 castration resistant prostate cancer (CRPC) cell line. The comparatively weak activities of series compounds in the H3-DHT and/or TIF2 box 3 LXXLL-peptide binding assays to the recombinant ligand binding domain of AR suggest that direct antagonism at the orthosteric ligand binding site or AF-2 surface respectively are unlikely mechanisms of action. Cellular enhanced thermal stability assays (CETSA) indicated that compounds engaged AR and reduced the maximum efficacy and right shifted the EC50 of DHT-enhanced AR thermal stabilization consistent with the effects of negative allosteric modulators. Molecular docking of potent representative hits from each series to AR structures suggest that S1–1 and S2–6 engage a novel binding pocket (BP-1) adjacent to the orthosteric ligand binding site, while S3–11 occupies the AR binding function 3 (BF-3) allosteric pocket. Hit binding poses indicate spaces and residues adjacent to the BP-1 and BF-3 pockets that will be exploited in future medicinal chemistry optimization studies. Small molecule allosteric modulators that prevent/disrupt AR PPIs with coactivators like TIF2 to alter transcriptional activation in the presence of orthosteric agonists might evade the resistance mechanisms to existing prostate cancer drugs and provide novel starting points for medicinal chemistry lead optimization and future development into therapies for metastatic CRPC. |
| first_indexed | 2024-03-11T15:18:50Z |
| format | Article |
| id | doaj.art-1f522784d4774c6bb7d40f6419a475d6 |
| institution | Directory Open Access Journal |
| issn | 2472-5552 |
| language | English |
| last_indexed | 2024-03-11T15:18:50Z |
| publishDate | 2023-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Discovery |
| spelling | doaj.art-1f522784d4774c6bb7d40f6419a475d62023-10-29T04:20:04ZengElsevierSLAS Discovery2472-55522023-10-01287325343Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancerAshley T. Fancher0Yun Hua1David A. Close2Wei Xu3Lee A. McDermott4Christopher J. Strock5Ulises Santiago6Carlos J. Camacho7Paul A. Johnston8Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA 15261, USA; Nucleus Global, 2 Ravinia Drive, Suite 605, Atlanta, GA 30346, USADepartment of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA 15261, USA; PsychoGenics Inc, 215 College Road, Paramus, NJ 07652, USACyprotex US, 313 Pleasant Street, Watertown, MA 02472, USADepartment of Computational and Systems Biology, School of Medicine, at the University of Pittsburgh, USADepartment of Computational and Systems Biology, School of Medicine, at the University of Pittsburgh, USADepartment of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA 15261, USA; University of Pittsburgh Hillman Cancer Center, Pittsburgh, PA 15232, USA; Corresponding author at: Professor, Department of Pharmaceutical Sciences, School of Pharmacy, Salk Hall Room 7402, 3501 Terrace Street, Pittsburgh PA 15261.Three series of compounds were prioritized from a high content screening campaign that identified molecules that blocked dihydrotestosterone (DHT) induced formation of Androgen Receptor (AR) protein-protein interactions (PPIs) with the Transcriptional Intermediary Factor 2 (TIF2) coactivator and also disrupted preformed AR-TIF2 PPI complexes; the hydrobenzo-oxazepins (S1), thiadiazol-5-piperidine-carboxamides (S2), and phenyl-methyl-indoles (S3). Compounds from these series inhibited AR PPIs with TIF2 and SRC-1, another p160 coactivator, in mammalian 2-hybrid assays and blocked transcriptional activation in reporter assays driven by full length AR or AR-V7 splice variants. Compounds inhibited the growth of five prostate cancer cell lines, with many exhibiting differential cytotoxicity towards AR positive cell lines. Representative compounds from the 3 series substantially reduced both endogenous and DHT-enhanced expression and secretion of the prostate specific antigen (PSA) cancer biomarker in the C4–2 castration resistant prostate cancer (CRPC) cell line. The comparatively weak activities of series compounds in the H3-DHT and/or TIF2 box 3 LXXLL-peptide binding assays to the recombinant ligand binding domain of AR suggest that direct antagonism at the orthosteric ligand binding site or AF-2 surface respectively are unlikely mechanisms of action. Cellular enhanced thermal stability assays (CETSA) indicated that compounds engaged AR and reduced the maximum efficacy and right shifted the EC50 of DHT-enhanced AR thermal stabilization consistent with the effects of negative allosteric modulators. Molecular docking of potent representative hits from each series to AR structures suggest that S1–1 and S2–6 engage a novel binding pocket (BP-1) adjacent to the orthosteric ligand binding site, while S3–11 occupies the AR binding function 3 (BF-3) allosteric pocket. Hit binding poses indicate spaces and residues adjacent to the BP-1 and BF-3 pockets that will be exploited in future medicinal chemistry optimization studies. Small molecule allosteric modulators that prevent/disrupt AR PPIs with coactivators like TIF2 to alter transcriptional activation in the presence of orthosteric agonists might evade the resistance mechanisms to existing prostate cancer drugs and provide novel starting points for medicinal chemistry lead optimization and future development into therapies for metastatic CRPC.http://www.sciencedirect.com/science/article/pii/S2472555223000539 |
| spellingShingle | Ashley T. Fancher Yun Hua David A. Close Wei Xu Lee A. McDermott Christopher J. Strock Ulises Santiago Carlos J. Camacho Paul A. Johnston Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancer SLAS Discovery |
| title | Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancer |
| title_full | Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancer |
| title_fullStr | Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancer |
| title_full_unstemmed | Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancer |
| title_short | Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation–Drug leads for metastatic castration resistant prostate cancer |
| title_sort | characterization of allosteric modulators that disrupt androgen receptor co activator protein protein interactions to alter transactivation drug leads for metastatic castration resistant prostate cancer |
| url | http://www.sciencedirect.com/science/article/pii/S2472555223000539 |
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