RÉSUMÉ
Patients with castration-resistant prostate cancer inevitably acquire resistance to antiandrogen therapies in part because of androgen receptor (AR) mutations or splice variants enabling restored AR signaling. Here we show that ligand-activated AR can form transcriptionally active condensates. Both structured and unstructured regions of AR contribute to the effective phase separation of AR and disordered N-terminal domain plays a predominant role. AR liquid-liquid phase separation behaviors faithfully report transcriptional activity and antiandrogen efficacy. Antiandrogens can promote phase separation and transcriptional activity of AR-resistant mutants in a ligand-independent manner. We conducted a phase-separation-based phenotypic screen and identified ET516 that specifically disrupts AR condensates, effectively suppresses AR transcriptional activity and inhibits the proliferation and tumor growth of prostate cancer cells expressing AR-resistant mutants. Our results demonstrate liquid-liquid phase separation as an emerging mechanism underlying drug resistance and show that targeting phase separation may provide a feasible approach for drug discovery.
Sujet(s)
Tumeurs prostatiques résistantes à la castration , Tumeurs de la prostate , Mâle , Humains , Récepteurs aux androgènes/génétique , Antagonistes des androgènes/pharmacologie , Antagonistes des androgènes/usage thérapeutique , Ligands , Résistance aux médicaments antinéoplasiques , Tumeurs de la prostate/traitement médicamenteux , Lignée cellulaire tumorale , Tumeurs prostatiques résistantes à la castration/traitement médicamenteux , Tumeurs prostatiques résistantes à la castration/génétique , Tumeurs prostatiques résistantes à la castration/anatomopathologieRÉSUMÉ
Polycomb Repressive Complex 2 (PRC2) plays an important role in transcriptional regulation during animal development and in cell differentiation, and alteration of PRC2 activity has been associated with cancer. On a molecular level, PRC2 catalyzes methylation of histone H3 lysine 27 (H3K27), resulting in mono-, di-, or trimethylated forms of H3K27, of which the trimethylated form H3K27me3 leads to transcriptional repression of polycomb target genes. Previously, we have shown that binding of the low-molecular-weight compound EED226 to the H3K27me3 binding pocket of the regulatory subunit EED can effectively inhibit PRC2 activity in cells and reduce tumor growth in mouse xenograft models. Here, we report the stepwise optimization of the tool compound EED226 toward the potent and selective EED inhibitor MAK683 (compound 22) and its subsequent preclinical characterization. Based on a balanced PK/PD profile, efficacy, and mitigated risk of forming reactive metabolites, MAK683 has been selected for clinical development.
Sujet(s)
Histone , Tumeurs , Animaux , Antienzymes , Histone/métabolisme , Humains , Méthylation , Souris , Tumeurs/traitement médicamenteux , Complexe répresseur Polycomb-2RÉSUMÉ
Polycomb repressive complex 2 (PRC2), a histone H3 lysine 27 methyltransferase, plays a key role in gene regulation and is a known epigenetics drug target for cancer therapy. The WD40 domain-containing protein EED is the regulatory subunit of PRC2. It binds to the tri-methylated lysine 27 of the histone H3 (H3K27me3), and through which stimulates the activity of PRC2 allosterically. Recently, we disclosed a novel PRC2 inhibitor EED226 which binds to the K27me3-pocket on EED and showed strong antitumor activity in xenograft mice model. Here, we further report the identification and validation of four other EED binders along with EED162, the parental compound of EED226. The crystal structures for all these five compounds in complex with EED revealed a common deep pocket induced by the binding of this diverse set of compounds. This pocket was created after significant conformational rearrangement of the aromatic cage residues (Y365, Y148 and F97) in the H3K27me3 binding pocket of EED, the width of which was delineated by the side chains of these rearranged residues. In addition, all five compounds interact with the Arg367 at the bottom of the pocket. Each compound also displays unique features in its interaction with EED, suggesting the dynamics of the H3K27me3 pocket in accommodating the binding of different compounds. Our results provide structural insights for rational design of novel EED binder for the inhibition of PRC2 complex activity.
Sujet(s)
Antienzymes/pharmacologie , Simulation de docking moléculaire , Complexe répresseur Polycomb-2/antagonistes et inhibiteurs , Sulfones/pharmacologie , Triazoles/pharmacologie , Animaux , Sites de fixation , Découverte de médicament , Antienzymes/composition chimique , Tests de criblage à haut débit , Souris , Complexe répresseur Polycomb-2/composition chimique , Complexe répresseur Polycomb-2/métabolisme , Relation quantitative structure-activité , Sulfones/composition chimique , Triazoles/composition chimiqueRÉSUMÉ
Overexpression and somatic heterozygous mutations of EZH2, the catalytic subunit of polycomb repressive complex 2 (PRC2), are associated with several tumor types. EZH2 inhibitor, EPZ-6438 (tazemetostat), demonstrated clinical efficacy in patients with acceptable safety profile as monotherapy. EED, another subunit of PRC2 complex, is essential for its histone methyltransferase activity through direct binding to trimethylated lysine 27 on histone 3 (H3K27Me3). Herein we disclose the discovery of a first-in-class potent, selective, and orally bioavailable EED inhibitor compound 43 (EED226). Guided by X-ray crystallography, compound 43 was discovered by fragmentation and regrowth of compound 7, a PRC2 HTS hit that directly binds EED. The ensuing scaffold hopping followed by multiparameter optimization led to the discovery of 43. Compound 43 induces robust and sustained tumor regression in EZH2MUT preclinical DLBCL model. For the first time we demonstrate that specific and direct inhibition of EED can be effective as an anticancer strategy.
Sujet(s)
Antinéoplasiques/composition chimique , Antinéoplasiques/pharmacologie , Complexe répresseur Polycomb-2/antagonistes et inhibiteurs , Sulfones/composition chimique , Sulfones/pharmacologie , Triazoles/composition chimique , Triazoles/pharmacologie , Animaux , Antinéoplasiques/pharmacocinétique , Antinéoplasiques/usage thérapeutique , Prolifération cellulaire/effets des médicaments et des substances chimiques , Cristallographie aux rayons X , Chiens , Femelle , Haplorhini , Histone/métabolisme , Humains , Lymphome B diffus à grandes cellules/traitement médicamenteux , Lymphome B diffus à grandes cellules/métabolisme , Lysine/métabolisme , Mâle , Méthylation/effets des médicaments et des substances chimiques , Souris , Souris de lignée BALB C , Souris nude , Simulation de docking moléculaire , Complexe répresseur Polycomb-2/composition chimique , Complexe répresseur Polycomb-2/métabolisme , Rats , Sulfones/pharmacocinétique , Sulfones/usage thérapeutique , Triazoles/pharmacocinétique , Triazoles/usage thérapeutiqueRÉSUMÉ
A series of trisubstituted hydroxylactams was identified as potent enzymatic and cellular inhibitors of human lactate dehydrogenase A. Utilizing structure-based design and physical property optimization, multiple inhibitors were discovered with <10 µM lactate IC50 in a MiaPaca2 cell line. Optimization of the series led to 29, a potent cell active molecule (MiaPaca2 IC50 = 0.67 µM) that also possessed good exposure when dosed orally to mice.
RÉSUMÉ
The 2A proteinase (2A(pro)) is an enterovirally encoded cysteine protease that plays essential roles in both the processing of viral precursor polyprotein and the hijacking of host cell translation and other processes in the virus life cycle. Crystallographic studies of 2A(pro) from enterovirus 71 (EV71) and its interaction with the substrate are reported here. EV71 2A(pro) was comprised of an N-terminal domain of a four-stranded antiparallel ß sheet and a C-terminal domain of a six-stranded antiparallel ß barrel with a tightly bound zinc atom. Unlike in other 2A(pro) structures, there is an open cleft across the surface of the protein in an open conformation. As demonstrated by the crystallographic studies and modeling of the complex structure, the open cleft could be fitted with the substrate. On comparison 2A(pro) of EV71 to those of the human rhinovirus 2 and coxsackievirus B4, the open conformation could be closed with a hinge motion in the bII2 and cII ß strands. This was supported by molecular dynamic simulation. The structural variation among different 2A(pro) structures indicates a conformational flexibility in the substrate-binding cleft. The open structure provides an accessible framework for the design and development of therapeutics against the viral target.
Sujet(s)
Cysteine endopeptidases/composition chimique , Entérovirus humain A/enzymologie , Modèles moléculaires , Conformation des protéines , Séquence d'acides aminés , Cristallographie aux rayons X , Cysteine endopeptidases/génétique , Escherichia coli , Transfert d'énergie par résonance de fluorescence , Vecteurs génétiques/génétique , Données de séquences moléculaires , Mutagenèse , Réaction de polymérisation en chaîne , Alignement de séquencesRÉSUMÉ
LBVS of 12480 in-house compounds, followed by HTRF assay, resulted in one nonsteroidal compound (11) with antagonistic activity against FXR (69.01 ± 11.75 µM). On the basis of 11, 26 new derivatives (12a-z) were designed and synthesized accordingly. Five derivatives (12f-g, 12p, 12u, and 12y) showed better antagonistic activities against FXR than compound 11. Remarkably, the most potent derivative, 12u (8.96 ± 3.62 µM), showed antagonistic capability approximately 10 times and 8-fold higher than that of the control (GS) and the starting compound 11, respectively. 12u was further confirmed to have high binding affinity with FXRαLBD, FXR specificity over six other nuclear receptors, and potent antagonistic activity against FXR in two cell testing platforms. 12u strongly suppressed the regulating effects of CDCA on FXR target genes. The therapeutic potential of 12u was identified by lowering the contents of triglyceride and cholesterol in human hepatoma HepG2 cells and in the cholesterol-fed C57BL/6 mices.
Sujet(s)
Anticholestérolémiants/synthèse chimique , Composés benzylidéniques/synthèse chimique , Pyrazoles/synthèse chimique , Pyrazolones/synthèse chimique , Récepteurs cytoplasmiques et nucléaires/antagonistes et inhibiteurs , Animaux , Anticholestérolémiants/composition chimique , Anticholestérolémiants/pharmacologie , Composés benzylidéniques/composition chimique , Composés benzylidéniques/pharmacologie , Biodisponibilité , Chénodiol/pharmacologie , Cholestérol/métabolisme , Bases de données chimiques , Cellules HEK293 , Cellules HepG2 , Humains , Foie/métabolisme , Mâle , Souris , Souris de lignée C57BL , Modèles moléculaires , Pyrazoles/composition chimique , Pyrazoles/pharmacologie , Pyrazolones/composition chimique , Pyrazolones/pharmacologie , Rats , Récepteurs cytoplasmiques et nucléaires/agonistes , Récepteurs cytoplasmiques et nucléaires/génétique , Solubilité , Stéréoisomérie , Relation structure-activité , Transcription génétique , Activation de la transcription , Triglycéride/métabolismeRÉSUMÉ
BACKGROUND: Infarct-induced left ventricular (LV) remodeling is a deleterious consequence after acute myocardial infarction (MI) which may further advance to congestive heart failure. Therefore, new therapeutic strategies to attenuate the effects of LV remodeling are urgently needed. Salvianolic acid B (SalB) from Salviae mitiorrhizae, which has been widely used in China for the treatment of cardiovascular diseases, is a potential candidate for therapeutic intervention of LV remodeling targeting matrix metalloproteinase-9 (MMP-9). RESULTS: Molecular modeling and LIGPLOT analysis revealed in silico docking of SalB at the catalytic site of MMP-9. Following this lead, we expressed truncated MMP-9 which contains only the catalytic domain, and used this active protein for in-gel gelatin zymography, enzymatic analysis, and SalB binding by Biacore. Data generated from these assays indicated that SalB functioned as a competitive inhibitor of MMP-9. In our rat model for cardiac remodeling, western blot, echocardiography, hemodynamic measurement and histopathological detection were used to detect the effects and mechanism of SalB on cardio-protection. Our results showed that in MI rat, SalB selectively inhibited MMP-9 activities without affecting MMP-9 expression while no effect of SalB was seen on MMP-2. Moreover, SalB treatment in MI rat could efficiently increase left ventricle wall thickness, improve heart contractility, and decrease heart fibrosis. CONCLUSIONS: As a competitive inhibitor of MMP-9, SalB presents significant effects on preventing LV structural damage and preserving cardiac function. Further studies to develop SalB and its analogues for their potential for cardioprotection in clinic are warranted.
Sujet(s)
Benzofuranes/pharmacologie , Benzofuranes/usage thérapeutique , Inhibiteurs de métalloprotéinases matricielles , Infarctus du myocarde/traitement médicamenteux , Inhibiteurs de protéases/pharmacologie , Inhibiteurs de protéases/usage thérapeutique , Remodelage ventriculaire/effets des médicaments et des substances chimiques , Animaux , Benzofuranes/composition chimique , Benzofuranes/métabolisme , Sites de fixation , Simulation numérique , Régulation négative/effets des médicaments et des substances chimiques , Conception de médicament , Médicaments issus de plantes chinoises/composition chimique , Médicaments issus de plantes chinoises/pharmacologie , Médicaments issus de plantes chinoises/usage thérapeutique , Fibrose , Ventricules cardiaques/effets des médicaments et des substances chimiques , Ventricules cardiaques/métabolisme , Ventricules cardiaques/anatomopathologie , Ventricules cardiaques/physiopathologie , Humains , Mâle , Matrix metalloproteinase 2/métabolisme , Matrix metalloproteinase 9/composition chimique , Matrix metalloproteinase 9/génétique , Matrix metalloproteinase 9/métabolisme , Conformation moléculaire , Contraction myocardique/effets des médicaments et des substances chimiques , Infarctus du myocarde/métabolisme , Infarctus du myocarde/anatomopathologie , Infarctus du myocarde/physiopathologie , Inhibiteurs de protéases/composition chimique , Inhibiteurs de protéases/métabolisme , Répartition aléatoire , Rats , Rat Wistar , Protéines de fusion recombinantes/antagonistes et inhibiteurs , Protéines de fusion recombinantes/composition chimique , Protéines de fusion recombinantes/génétique , Protéines de fusion recombinantes/métabolismeRÉSUMÉ
The severe acute respiratory syndrome (SARS) coronavirus 3CL protease is an attractive target for the development of anti-SARS drugs. In this paper, cinanserin (1) analogs were synthesized and tested for the inhibitory activities against SARS-coronavirus (CoV) 3CL protease by fluorescence resonance energy transfer (FRET) assay. Four analogs show significant activities, especially compound 26 with an IC(50) of 1.06 microM.
Sujet(s)
Antiviraux/pharmacologie , Cinansérine/analogues et dérivés , Cinansérine/pharmacologie , Inhibiteurs de protéases/pharmacologie , Antisérotonines/pharmacologie , Virus du SRAS/enzymologie , Protéines virales/antagonistes et inhibiteurs , Antiviraux/synthèse chimique , Cinansérine/synthèse chimique , Protéases 3C des coronavirus , Cysteine endopeptidases , Conception de médicament , Transfert d'énergie par résonance de fluorescence , Indicateurs et réactifs , Spectroscopie par résonance magnétique , Modèles moléculaires , Inhibiteurs de protéases/synthèse chimique , Antisérotonines/synthèse chimique , Spectrophotométrie IR , Relation structure-activitéRÉSUMÉ
BACKGROUND: Target identification is important for modern drug discovery. With the advances in the development of molecular docking, potential binding proteins may be discovered by docking a small molecule to a repository of proteins with three-dimensional (3D) structures. To complete this task, a reverse docking program and a drug target database with 3D structures are necessary. To this end, we have developed a web server tool, TarFisDock (Target Fishing Docking) http://www.dddc.ac.cn/tarfisdock, which has been used widely by others. Recently, we have constructed a protein target database, Potential Drug Target Database (PDTD), and have integrated PDTD with TarFisDock. This combination aims to assist target identification and validation. DESCRIPTION: PDTD is a web-accessible protein database for in silico target identification. It currently contains >1100 protein entries with 3D structures presented in the Protein Data Bank. The data are extracted from the literatures and several online databases such as TTD, DrugBank and Thomson Pharma. The database covers diverse information of >830 known or potential drug targets, including protein and active sites structures in both PDB and mol2 formats, related diseases, biological functions as well as associated regulating (signaling) pathways. Each target is categorized by both nosology and biochemical function. PDTD supports keyword search function, such as PDB ID, target name, and disease name. Data set generated by PDTD can be viewed with the plug-in of molecular visualization tools and also can be downloaded freely. Remarkably, PDTD is specially designed for target identification. In conjunction with TarFisDock, PDTD can be used to identify binding proteins for small molecules. The results can be downloaded in the form of mol2 file with the binding pose of the probe compound and a list of potential binding targets according to their ranking scores. CONCLUSION: PDTD serves as a comprehensive and unique repository of drug targets. Integrated with TarFisDock, PDTD is a useful resource to identify binding proteins for active compounds or existing drugs. Its potential applications include in silico drug target identification, virtual screening, and the discovery of the secondary effects of an old drug (i.e. new pharmacological usage) or an existing target (i.e. new pharmacological or toxic relevance), thus it may be a valuable platform for the pharmaceutical researchers. PDTD is available online at http://www.dddc.ac.cn/pdtd/.
Sujet(s)
Bases de données de protéines , Systèmes de délivrance de médicaments/méthodes , Conception de médicament , Internet , Protéines/composition chimique , Protéines/ultrastructure , Interface utilisateur , Sites de fixation , Systèmes de gestion de bases de données , Mémorisation et recherche des informations/méthodes , Liaison aux protéines , Conformation des protéines , Analyse de séquence de protéine/méthodesRÉSUMÉ
Deoxyelephantopin (ESD) was reported to potentiate apoptosis, inhibit invasion and abolish osteoclastogenesis but no target protein was disclosed. Here, we discovered that ESD could significantly inhibit the proliferation of different cancer cells and induce apoptosis and cell cycle arrest at G(2)/M phase in HeLa cell. Moreover, biochemical and biophysical assays revealed that ESD acted as a specific partial agonist against PPARgamma. Molecular docking with site-directed mutagenesis analyses indicated that ESD functioned as a partial agonist of PPARgamma by adopting a distinct binding mode to PPARgamma compared with rosiglitazone. The PPARgamma knockdown results indicated that the inhibition of ESD against the cancer cell proliferation is more possibly through PPARgamma-independent pathway and our findings might supply potent binding features for ESD/PPARgamma interaction at atomic level, and shed light on the potential acting target information for this natural compound.
Sujet(s)
Antinéoplasiques/pharmacologie , Lactones/pharmacologie , Récepteur PPAR gamma/agonistes , Sesquiterpènes/pharmacologie , Animaux , Antinéoplasiques/métabolisme , Apoptose , Cellules COS , Cycle cellulaire/effets des médicaments et des substances chimiques , Lignée cellulaire tumorale , Prolifération cellulaire/effets des médicaments et des substances chimiques , Chlorocebus aethiops , Cellules HeLa , Humains , Lactones/métabolisme , Modèles moléculaires , Récepteur PPAR gamma/métabolisme , Sesquiterpènes/métabolismeRÉSUMÉ
TarFisDock is a web-based tool for automating the procedure of searching for small molecule-protein interactions over a large repertoire of protein structures. It offers PDTD (potential drug target database), a target database containing 698 protein structures covering 15 therapeutic areas and a reverse ligand-protein docking program. In contrast to conventional ligand-protein docking, reverse ligand-protein docking aims to seek potential protein targets by screening an appropriate protein database. The input file of this web server is the small molecule to be tested, in standard mol2 format; TarFisDock then searches for possible binding proteins for the given small molecule by use of a docking approach. The ligand-protein interaction energy terms of the program DOCK are adopted for ranking the proteins. To test the reliability of the TarFisDock server, we searched the PDTD for putative binding proteins for vitamin E and 4H-tamoxifen. The top 2 and 10% candidates of vitamin E binding proteins identified by TarFisDock respectively cover 30 and 50% of reported targets verified or implicated by experiments; and 30 and 50% of experimentally confirmed targets for 4H-tamoxifen appear amongst the top 2 and 5% of the TarFisDock predicted candidates, respectively. Therefore, TarFisDock may be a useful tool for target identification, mechanism study of old drugs and probes discovered from natural products. TarFisDock and PDTD are available at http://www.dddc.ac.cn/tarfisdock/.