Discovery of potent BET bromodomain 1 stereoselective inhibitors using DNA-encoded chemical library selections.
Discovery of potent BET bromodomain 1 stereoselective inhibitors using DNA-encoded chemical library selections.
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DOI：
10.1073/pnas.2122506119
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发表时间：
2022-05-31
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
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作者：
BET溴域抑制对多种疾病有治疗作用；然而，泛BET抑制剂诱导了显著的骨髓抑制和胃肠道毒性，可能是由于抑制了所有BET家族成员的串联溴结构域（BD）。然而，在癌症和其他疾病的临床前模型中，选择性抑制第一种BD （BD1）表型会影响泛β - bet抑制剂的活性，对于BRDT，在睾丸中具有避孕作用。在这里，我们利用数十亿分子的dna编码化学文库（decl）来鉴定具有高效、稳定和良好细胞活性的特定手性的BET bd1选择性抑制剂。我们的研究结果强调了DECL平台在识别特异性、强效蛋白结合物方面的稳健性和有效性，这些蛋白结合物有望成为潜在的抗癌和抗炎剂以及男性避孕药。BRDT、BRD2、BRD3和BRD4组成了含有两个相似串联溴域（BD1和BD2）的溴域和外域（BET）亚家族。选择性BD1抑制表型表明，在癌症模型中，以及在睾丸中，正如我们和其他人在BRDT中所报道的那样，串联BET - BD抑制的效果。为了寻找新的BET -BD1结合物，我们从dna编码的化学文库中与BRDT-BD1或BRDT-BD2蛋白平行筛选了bb45亿个分子。我们在dna外重新合成了仅由BRDT-BD1富集的化合物系列，发现了一个有效的手性化合物CDD-724，其抑制BRDT-BD1的选择性是BRDT-BD2的2000倍。CDD-724立体异构体对BRDT-BD1的抑制作用差异显著，其中r -对映体（CDD-787）比s -对映体（CDD-786）强50倍。通过结构-活性关系研究，我们生产出了CDD-956，它保持了皮摩尔BET BD1的结合能力和对BET BD2蛋白的高选择性，并且比CDD-787在人肝微粒体中具有更高的稳定性。BROMOscan分析证实了CDD-787和CDD-956对BD1具有良好的pan-BET亲和力和选择性，而不是BD2和所有其他含BD1的蛋白。BRDT-BD1与CDD-956结合的共晶结构为1.82 Å，发现BRDT-BD1与αZ和αC螺旋具有特异性接触，这解释了BET -BD1相对于BD2具有高亲和力和选择性。在NanoBRET实验中，CDD-787和CDD-956维持了细胞bd1的选择性，并在急性髓性白血病细胞系中显示出有效的抗白血病活性。这些BET bd1特异性和高效的化合物结构独特，为实现BET特异性提供了手性的重要性。
BET bromodomain inhibition is therapeutic in multiple diseases; however, pan-BET inhibitors have induced significant myelosuppression and gastrointestinal toxicity, perhaps due to inhibition of both tandem bromodomains (BD) of all BET family members. However, selective inhibition of just the first BD (BD1) phenocopies pan-BET inhibitor activity in preclinical models of cancer, other diseases, and, for BRDT, in the testes for a contraceptive effect. Here, we leveraged our multibillion-molecule collection of DNA-encoded chemical libraries (DECLs) to identify BET BD1-selective inhibitors of specific chirality with high potency, stability, and good cellular activity. Our findings highlight the robustness and efficiency of the DECL platform to identify specific, potent protein binders that have promise as potential anticancer and anti-inflammatory agents and as male contraceptives. BRDT, BRD2, BRD3, and BRD4 comprise the bromodomain and extraterminal (BET) subfamily which contain two similar tandem bromodomains (BD1 and BD2). Selective BD1 inhibition phenocopies effects of tandem BET BD inhibition both in cancer models and, as we and others have reported of BRDT, in the testes. To find novel BET BD1 binders, we screened >4.5 billion molecules from our DNA-encoded chemical libraries with BRDT-BD1 or BRDT-BD2 proteins in parallel. A compound series enriched only by BRDT-BD1 was resynthesized off-DNA, uncovering a potent chiral compound, CDD-724, with >2,000-fold selectivity for inhibiting BRDT-BD1 over BRDT-BD2. CDD-724 stereoisomers exhibited remarkable differences in inhibiting BRDT-BD1, with the R-enantiomer (CDD-787) being 50-fold more potent than the S-enantiomer (CDD-786). From structure–activity relationship studies, we produced CDD-956, which maintained picomolar BET BD1 binding potency and high selectivity over BET BD2 proteins and had improved stability in human liver microsomes over CDD-787. BROMOscan profiling confirmed the excellent pan-BET BD1 affinity and selectivity of CDD-787 and CDD-956 on BD1 versus BD2 and all other BD-containing proteins. A cocrystal structure of BRDT-BD1 bound with CDD-956 was determined at 1.82 Å and revealed BRDT-BD1–specific contacts with the αZ and αC helices that explain the high affinity and selectivity for BET BD1 versus BD2. CDD-787 and CDD-956 maintain cellular BD1-selectivity in NanoBRET assays and show potent antileukemic activity in acute myeloid leukemia cell lines. These BET BD1-specific and highly potent compounds are structurally unique and provide insight into the importance of chirality to achieve BET specificity.
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选择性抑制BET溴结构域。
影响因子：
64.8
作者：
通讯作者：
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DOI：
10.1126/science.aaz8455
发表时间：
2020-04-24
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gilan O;Rioja I;Knezevic K;Bell MJ;Yeung MM;Harker NR;Lam EYN;Chung CW;Bamborough P;Petretich M;Urh M;Atkinson SJ;Bassil AK;Roberts EJ;Vassiliadis D;Burr ML;Preston AGS;Wellaway C;Werner T;Gray JR;Michon AM;Gobbetti T;Kumar V;Soden PE;Haynes A;Vappiani J;Tough DF;Taylor S;Dawson SJ;Bantscheff M;Lindon M;Drewes G;Demont EH;Daniels DL;Grandi P;Prinjha RK;Dawson MA
通讯作者：
Dawson MA
影响因子：
64.5
作者：
Matzuk MM;McKeown MR;Filippakopoulos P;Li Q;Ma L;Agno JE;Lemieux ME;Picaud S;Yu RN;Qi J;Knapp S;Bradner JE
通讯作者：
Bradner JE
影响因子：
2.7
作者：
Berkovits BD;Wolgemuth DJ
通讯作者：
Wolgemuth DJ
DOI：
10.3390/v5061571
发表时间：
2013-06-21
期刊：
Viruses
影响因子：
--
作者：
Boehm D;Conrad RJ;Ott M
通讯作者：
Ott M