Discovery and characterization of bromodomain 2-specific inhibitors of BRDT
| Autor: | Justin L. Anglin, Kevin Riehle, John C. Faver, Marta Storl-Desmond, Rajesh Sharma, Damian W. Young, Melek N. Ucisik, Gundeep Kaur, Feng Li, Choel Kim, Kiran L. Sharma, Nicholas Simmons, Zhifeng Yu, Sureshbabu Nagarajan, Martin M. Matzuk, Pranavanand Nyshadham, Stephen Palmer, Banumathi Sankaran, Angela F. Ku |
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| Rok vydání: | 2021 |
| Předmět: |
Protein Conformation
alpha-Helical Male Subfamily Protein Conformation Gene Expression Quantitative Structure-Activity Relationship Cell Cycle Proteins Crystallography X-Ray Ligands 01 natural sciences Biochemistry Mice Testis Drug Discovery Cloning Molecular Cancer 0303 health sciences Multidisciplinary Crystallography Chemistry Nuclear Proteins Azepines Biological Sciences Small molecule Recombinant Proteins Molecular Docking Simulation small-molecule inhibitors Knockout mouse Protein Binding BRD4 Genetic Vectors 03 medical and health sciences DNA-encoded chemistry Contraceptive Agents In vivo Escherichia coli Genetics Potency Animals Humans Protein Interaction Domains and Motifs 030304 developmental biology BET bromodomains Binding Sites 010405 organic chemistry Contraception/Reproduction alpha-Helical Contraceptive Agents Male Molecular Triazoles In vitro 0104 chemical sciences Bromodomain High-Throughput Screening Assays male contraceptive X-Ray Protein Conformation beta-Strand beta-Strand Transcription Factors Cloning |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America, vol 118, iss 9 Proceedings of the National Academy of Sciences of the United States of America |
| Popis: | Significance There is no nonhormonal contraceptive pill for men, although hundreds of genes have been identified to play roles during spermatogenesis and fertilization in the male reproductive tract. To address the absence of contraceptive drugs for men, we established a DNA-encoded chemistry technology (DEC-Tec) platform. Our drug discovery campaign on BRDT, a validated spermatogenic-specific contraceptive target, yielded rapid discovery of potent and specific inhibitors of the second bromodomain of BRDT that have unique binding characteristics to BRDT-BD2 relative to BRDT-BD1. Our study emphasizes the robustness and validation of the DEC-Tec platform where the obtained structure–affinity relationship data would allow us to identify specific protein binders immediately without performing exhaustive medicinal chemistry optimization of compounds with potential as male contraceptives. Bromodomain testis (BRDT), a member of the bromodomain and extraterminal (BET) subfamily that includes the cancer targets BRD2, BRD3, and BRD4, is a validated contraceptive target. All BET subfamily members have two tandem bromodomains (BD1 and BD2). Knockout mice lacking BRDT-BD1 or both bromodomains are infertile. Treatment of mice with JQ1, a BET BD1/BD2 nonselective inhibitor with the highest affinity for BRD4, disrupts spermatogenesis and reduces sperm number and motility. To assess the contribution of each BRDT bromodomain, we screened our collection of DNA-encoded chemical libraries for BRDT-BD1 and BRDT-BD2 binders. High-enrichment hits were identified and resynthesized off-DNA and examined for their ability to compete with JQ1 in BRDT and BRD4 bromodomain AlphaScreen assays. These studies identified CDD-1102 as a selective BRDT-BD2 inhibitor with low nanomolar potency and >1,000-fold selectivity over BRDT-BD1. Structure–activity relationship studies of CDD-1102 produced a series of additional BRDT-BD2/BRD4-BD2 selective inhibitors, including CDD-1302, a truncated analog of CDD-1102 with similar activity, and CDD-1349, an analog with sixfold selectivity for BRDT-BD2 versus BRD4-BD2. BROMOscan bromodomain profiling confirmed the great affinity and selectivity of CDD-1102 and CDD-1302 on all BET BD2 versus BD1 with the highest affinity for BRDT-BD2. Cocrystals of BRDT-BD2 with CDD-1102 and CDD-1302 were determined at 2.27 and 1.90 Å resolution, respectively, and revealed BRDT-BD2 specific contacts that explain the high affinity and selectivity of these compounds. These BD2-specific compounds and their binding to BRDT-BD2 are unique compared with recent reports and enable further evaluation of their nonhormonal contraceptive potential in vitro and in vivo. |
| Databáze: | OpenAIRE |
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