TY  - JOUR
AB  - Microglia, the innate immune cells of the central nervous system (CNS), act as first responders to brain injury. Their ability to switch between different neuroprotective and neurotoxic phenotypes, plays a central role in maintaining brain homeostasis. Recently, the P2Y12 receptor (P2Y12R) has been identified as a promising molecular biomarker for microglia activity, as its expression level is dependent on microglia phenotype and function. P2Y12R positron emission tomography (PET) might be a valuable diagnostic tool, however, tracers with sufficient brain retention have not been reported so far. Herein, we report a brain-permeable P2Y12R PET tracer for in vivo imaging of P2Y12R-positive microglia. Nicotinate [18F]12 exhibited nanomolar affinity and specificity for the target receptor and showed a reduced uptake in microglia-depleted (PLX) mice, in comparison to WT and Trem2 knockout (Trem2-/-) mice. Ex vivo immunohistochemistry (IHC) and PET data revealed a strong correlation between microglia abundance, P2Y12R expression levels and tracer uptake.
AU  - Joseph, E.K.*
AU  - Kunze, L.H.*
AU  - Schaefer, R.*
AU  - Palumbo, G.*
AU  - Kugelmann, B.*
AU  - Wagner, S.*
AU  - Lammich, S.*
AU  - Feederle, R.
AU  - Willem, M.*
AU  - Werner, R.A.*
AU  - Brendel, M.*
AU  - Lindner, S.*
C1  - 75248
C2  - 57879
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 15543-15562
TI  - Design, synthesis and preclinical evaluation of a brain-permeable PET tracer for P2Y12 receptor imaging in the brain.
JO  - J. Med. Chem.
VL  - 68
IS  - 15
PB  - Amer Chemical Soc
PY  - 2025
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Emerging RNA viruses, including SARS-CoV-2, continue to be a major threat. Cell entry of SARS-CoV-2 particles via the endosomal pathway involves cysteine cathepsins. Due to ubiquitous expression, cathepsin L (CatL) is considered a promising drug target in the context of different viral and lysosome-related diseases. We characterized the anti-SARS-CoV-2 activity of a set of carbonyl- and succinyl epoxide-based inhibitors, which were previously identified as inhibitors of cathepsins or related cysteine proteases. Calpain inhibitor XII, MG-101, and CatL inhibitor IV possess antiviral activity in the very low nanomolar EC50 range in Vero E6 cells and inhibit CatL in the picomolar Ki range. We show a relevant off-target effect of CatL inhibition by the coronavirus main protease α-ketoamide inhibitor 13b. Crystal structures of CatL in complex with 14 compounds at resolutions better than 2 Å present a solid basis for structure-guided understanding and optimization of CatL inhibitors toward protease drug development.
AU  - Falke, S.*
AU  - Lieske, J.*
AU  - Herrmann, A.
AU  - Loboda, J.*
AU  - Karničar, K.*
AU  - Günther, S.*
AU  - Reinke, P.Y.A.*
AU  - Ewert, W.*
AU  - Usenik, A.*
AU  - Lindič, N.*
AU  - Sekirnik, A.*
AU  - Dretnik, K.*
AU  - Tsuge, H.*
AU  - Turk, V.*
AU  - Chapman, H.N.*
AU  - Hinrichs, W.*
AU  - Ebert, G.
AU  - Turk, D.*
AU  - Meents, A.*
C1  - 70540
C2  - 55654
SP  - 7048-7067
TI  - Structural elucidation and antiviral activity of covalent cathepsin L inhibitors.
JO  - J. Med. Chem.
VL  - 67
IS  - 9
PY  - 2024
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - A clinical casein kinase 2 inhibitor, CX-4945 (silmitasertib), shows significant affinity toward the DYRK1A and GSK3β kinases, involved in down syndrome phenotypes, Alzheimer's disease, circadian clock regulation, and diabetes. This off-target activity offers an opportunity for studying the effect of the DYRK1A/GSK3β kinase system in disease biology and possible line extension. Motivated by the dual inhibition of these kinases, we solved and analyzed the crystal structures of DYRK1A and GSK3β with CX-4945. We built a quantum-chemistry-based model to rationalize the compound affinity for CK2α, DYRK1A, and GSK3β kinases. Our calculations identified a key element for CK2α's subnanomolar affinity to CX-4945. The methodology is expandable to other kinase selectivity modeling. We show that the inhibitor limits DYRK1A- and GSK3β-mediated cyclin D1 phosphorylation and reduces kinase-mediated NFAT signaling in the cell. Given the CX-4945's clinical and pharmacological profile, this inhibitory activity makes it an interesting candidate with potential for application in additional disease areas.
AU  - Grygier, P.*
AU  - Pustelny, K.*
AU  - Nowak, J.*
AU  - Golik, P.*
AU  - Popowicz, G.M.
AU  - Plettenburg, O.
AU  - Dubin, G.*
AU  - Cardoso Micu Menezes, F.M.
AU  - Czarna, A.*
C1  - 67752
C2  - 54062
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 4009-4024
TI  - Silmitasertib (CX-4945), a clinically used CK2-kinase inhibitor with additional effects on GSK3β and DYRK1A kinases: A structural perspective.
JO  - J. Med. Chem.
VL  - 66
IS  - 6
PB  - Amer Chemical Soc
PY  - 2023
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Here, we report the fragment-based drug discovery of potent and selective fragments that disrupt the Spire2-FMN2 but not the Spire1-FMN2 interaction. Hit fragments were identified in a differential scanning fluorimetry-based screen of an in-house library of 755 compounds and subsequently validated in multiple orthogonal biophysical assays, including fluorescence polarization, microscale thermophoresis, and 1H-15N HSQC nuclear magnetic resonance. Extensive structure-activity relationships combined with molecular docking followed by chemical optimization led to the discovery of compound 13, which exhibits micromolar potency and high ligand efficiency (LE = 0.38). Therefore, this fragment represents a validated starting point for the future development of selective chemical probes targeting the Spire2-FMN2 interaction.
AU  - Kitel, R.*
AU  - Surmiak, E.*
AU  - Borggräfe, J.
AU  - Kalinowska-Tluscik, J.*
AU  - Golik, P.*
AU  - Czub, M.*
AU  - Uzar, W.*
AU  - Musielak, B.*
AU  - Madej, M.G.*
AU  - Popowicz, G.M.
AU  - Dubin, G.*
AU  - Holak, T.A.*
C1  - 68881
C2  - 53736
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 15715-15727
TI  - Discovery of inhibitory fragments that selectively target spire2-FMN2 interaction.
JO  - J. Med. Chem.
VL  - 66
IS  - 23
PB  - Amer Chemical Soc
PY  - 2023
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Targeting the protein-protein interaction (PPI) between the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) and its repressor, Kelch-like ECH-associated protein 1 (Keap1), constitutes a promising strategy for treating diseases involving oxidative stress and inflammation. Here, a fragment-based drug discovery (FBDD) campaign resulted in novel, high-affinity (Ki = 280 nM), and cell-active noncovalent small-molecule Keap1-Nrf2 PPI inhibitors. We screened 2500 fragments using orthogonal assays─fluorescence polarization (FP), thermal shift assay (TSA), and surface plasmon resonance (SPR)─and validated the hits by saturation transfer difference (STD) NMR, leading to 28 high-priority hits. Thirteen co-structures showed fragments binding mainly in the P4 and P5 subpockets of Keap1's Kelch domain, and three fluorenone-based fragments featuring a novel binding mode were optimized by structure-based drug discovery. We thereby disclose several fragment hits, including their binding modes, and show how FBDD can be performed to find new small-molecule Keap1-Nrf2 PPI inhibitors.
AU  - Narayanan, D.*
AU  - Tran, K.T.*
AU  - Pallesen, J.S.*
AU  - Solbak, S.M.*
AU  - Qin, Y.*
AU  - Mukminova, E.*
AU  - Luchini, M.*
AU  - Vasilyeva, K.O.*
AU  - González Chichón, D.*
AU  - Goutsiou, G.*
AU  - Poulsen, C.*
AU  - Haapanen, N.*
AU  - Popowicz, G.M.
AU  - Sattler, M.
AU  - Olagnier, D.*
AU  - Gajhede, M.*
AU  - Bach, A.*
C1  - 66474
C2  - 53186
SP  - 14481-14526
TI  - Development of noncovalent small-molecule Keap1-Nrf2 inhibitors by fragment-based drug discovery.
JO  - J. Med. Chem.
VL  - 65
IS  - 21
PY  - 2022
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - A morpholine-based nucleotide analog was developed as a building block for hepatic siRNA targeting and stabilization. Attachment of an asialoglycoprotein-binding GalNAc ligand at the morpholine nitrogen was realized with different linkers. The obtained morpholino GalNAc scaffolds were coupled to the sense strand of a transthyretin-targeting siRNA and tested for their knockdown potency in vitro and in vivo. A clear structure-activity relationship was developed with regard to the linker type and length as well as the attachment site of the morpholino GalNAc moieties at the siRNA sense strand. Further, simple alkylation of the morpholine nitrogen led to a nucleotide analog, which increased siRNA stability, when used as a double 3'-overhang at the sense strand sequence. Combination of the best morpholino GalNAc building blocks as targeting nucleotides with an optimized stabilizing alkyl-substituted morpholine as 3'-overhangs resulted in siRNAs without any phosphorothioate stabilization in the sense strand and clearly improved the duration of action in vivo.
AU  - Hofmeister, A.*
AU  - Jahn-Hofmann, K.*
AU  - Brunner, B.*
AU  - Helms, M.W.*
AU  - Metz-Weidmann, C.*
AU  - Krack, A.*
AU  - Kurz, M.*
AU  - Li, Z.*
AU  - Weitzenberg, M.M.
AU  - Pflimlin, E.*
AU  - Plettenburg, O.
AU  - Scheidler, S.*
C1  - 61939
C2  - 50522
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 6838–6855
TI  - Syntheses of morpholine-based nucleotide analogs for hepatic siRNA targeting and stabilization.
JO  - J. Med. Chem.
VL  - 64
IS  - 10
PB  - Amer Chemical Soc
PY  - 2021
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Targeting the protein-protein interaction (PPI) between nuclear factor erythroid 2-related factor 2 (Nrf2) and Kelch-like ECH-associated protein 1 (Keap1) is a potential therapeutic strategy to control diseases involving oxidative stress. Here, six classes of known small-molecule Keap1-Nrf2 PPI inhibitors were dissected into 77 fragments in a fragment-based deconstruction reconstruction (FBDR) study and tested in four orthogonal assays. This gave 17 fragment hits of which six were shown by X-ray crystallography to bind in the Keap1 Kelch binding pocket. Two hits were merged into compound 8 with a 220-380-fold stronger affinity (Ki = 16 μM) relative to the parent fragments. Systematic optimization resulted in several novel analogues with Ki values of 0.04-0.5 μM, binding modes determined by X-ray crystallography, and enhanced microsomal stability. This demonstrates how FBDR can be used to find new fragment hits, elucidate important ligand-protein interactions, and identify new potent inhibitors of the Keap1-Nrf2 PPI.
AU  - Pallesen, J.S.*
AU  - Narayanan, D.*
AU  - Tran, K.T.*
AU  - Solbak, S.M.*
AU  - Marseglia, G.*
AU  - Sørensen, L.M.E.*
AU  - Høj, L.J.*
AU  - Munafò, F.*
AU  - Carmona, R.M.C.*
AU  - Garcia, A.D.*
AU  - Desu, H.L.*
AU  - Brambilla, R.*
AU  - Johansen, T.N.*
AU  - Popowicz, G.M.
AU  - Sattler, M.
AU  - Gajhede, M.*
AU  - Bach, A.*
C1  - 61730
C2  - 50415
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 4623-4661
TI  - Deconstructing noncovalent kelch-like ECH-associated protein 1 (Keap1) inhibitors into fragments to reconstruct new potent compounds.
JO  - J. Med. Chem.
VL  - 64
IS  - 8
PB  - Amer Chemical Soc
PY  - 2021
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Trypanosoma protists are pathogens leading to a spectrum of devastating infectious diseases. The range of available chemotherapeutics against Trypanosoma is limited, and the existing therapies are partially ineffective and cause serious adverse effects. Formation of the PEX14-PEX5 complex is essential for protein import into the parasites' glycosomes. This transport is critical for parasite metabolism and failure leads to mislocalization of glycosomal enzymes, with fatal consequences for the parasite. Hence, inhibiting the PEX14-PEX5 protein protein interaction (PPI) is an attractive way to affect multiple metabolic pathways. Herein, we have used structure-guided computational screening and optimization to develop the first line of compounds that inhibit PEX14-PEX5 PPI. The optimization was driven by several X-ray structures, NMR binding data, and molecular dynamics simulations. Importantly, the developed compounds show significant cellular activity against Trypanosoma, including the human pathogen Trypanosoma brucei gambiense and Trypanosoma cruzi parasites.
AU  - Dawidowski, M.
AU  - Kalel, V.C.*
AU  - Napolitano, V.*
AU  - Fino, R.
AU  - Schorpp, K.K.
AU  - Emmanouilidis, L.
AU  - Lenhart, D.
AU  - Ostertag, M.S.
AU  - Kaiser, M.*
AU  - Kolonko, M.
AU  - Tippler, B.*
AU  - Schliebs, W.*
AU  - Dubin, G.*
AU  - Mäser, P.*
AU  - Tetko, I.V.
AU  - Hadian, K.
AU  - Plettenburg, O.
AU  - Erdmann, R.*
AU  - Sattler, M.
AU  - Popowicz, G.M.
C1  - 57992
C2  - 48020
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 847-879
TI  - Structure-activity relationship in pyrazolo[4,3-c]pyridines, first inhibitors of PEX14-PEX5 Protein-Protein Interaction (PPI) with trypanocidal activity.
JO  - J. Med. Chem.
VL  - 63
IS  - 2
PB  - Amer Chemical Soc
PY  - 2020
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - The serine protease kallikrein-related peptidase 7 (KLK7) is a member of the human tissue kallikreins. Its dysregulation leads to pathophysiological inflammatory processes in the skin. Furthermore, it plays a role in several types of cancer. For the treatment of KLK7-associated diseases, coumarinic esters have been developed as small-molecule enzyme inhibitors. To characterize the inhibition mode of these inhibitors, we analyzed structures of the inhibited protease by X-ray crystallography. Electron density shows the inhibitors covalently attached to His57 of the catalytic triad. This confirms the irreversible character of the inhibition process. Upon inhibitor binding, His57 undergoes an outward rotation; thus, the catalytic triad of the protease is disrupted. Besides, the halophenyl moiety of the inhibitor was absent in the final enzyme-inhibitor complex due to the hydrolysis of the ester linkage. With these results, we analyze the structural basis of KLK7 inhibition by the covalent attachment of aromatic coumarinic esters.
AU  - Hanke, S.*
AU  - Tindall, C.*
AU  - Pippel, J.*
AU  - Ulbricht, D.*
AU  - Pirotte, B.*
AU  - Reboud-Ravaux, M.*
AU  - Heiker, J.T.
AU  - Sträter, N.*
C1  - 59030
C2  - 48485
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 5723-5733
TI  - Structural studies on the inhibitory binding mode of aromatic coumarinic esters to human kallikrein-related peptidase 7.
JO  - J. Med. Chem.
VL  - 63
IS  - 11
PB  - Amer Chemical Soc
PY  - 2020
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Nicotinamide adenine dinucleotide phosphate oxidase isoform 2 is an enzyme complex, which generates reactive oxygen species and contributes to oxidative stress. The p47phox-p22phox interaction is critical for the activation of the catalytical NOX2 domain, and p47phox is a potential target for therapeutic intervention. By screening 2500 fragments using fluorescence polarization and a thermal shift assay and validation by surface plasmon resonance, we found eight hits toward the tandem SH3 domain of p47phox (p47phox(SH3A-B)) with K-D values of 400-600 mu M. Structural studies revealed that fragments 1 and 2 bound two separate binding sites in the elongated conformation of p47phox(SH3A-B) and these competed with p22phox for binding to p47phox(SH3A-B). Chemical optimization led to a dimeric compound with the ability to potently inhibit the p47phox(SH3A-B)-p22phox interaction (K-i of 20 mu M). Thereby, we reveal a new way of targeting p47phox and present the first report of drug-like molecules with the ability to bind p47phox and inhibit its interaction with p22phox.
AU  - Solbak, S.M.*
AU  - Zang, J.*
AU  - Narayanan, D.*
AU  - Høj, L.J.*
AU  - Bucciarelli, S.*
AU  - Softley, C.
AU  - Meier, S.*
AU  - Langkilde, A.E.*
AU  - Gotfredsen, C.H.*
AU  - Sattler, M.
AU  - Bach, A.*
C1  - 57833
C2  - 48087
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 1156-1177
TI  - Developing inhibitors of the p47phox-p22phox protein-protein interaction by fragment-based drug discovery.
JO  - J. Med. Chem.
VL  - 63
IS  - 3
PB  - Amer Chemical Soc
PY  - 2020
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Constitutive proteolytic activity of MALT1 is associated with highly aggressive B-cell lymphomas. Chemical tools that detect active MALT1 have been reported, but suffer from poor cell permeability and/or cross-reactivity with the cysteine protease cathepsin B. Here, we report that the non-natural amino acid pipecolinic acid in the P2 position of substrates and chemical probes leads to improved selectivity toward MALT1 and results in cell-permeable fluorescent probes.
AU  - van de Plassche, M.A.T.*
AU  - O'Neill, T.J.
AU  - Seeholzer, T.
AU  - Turk, B.*
AU  - Krappmann, D.
AU  - Verhelst, S.H.L.*
C1  - 59180
C2  - 48615
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 3996-4004
TI  - Use of non-natural amino acids for the design and synthesis of a selective, cell-permeable MALT1 activity-based probe.
JO  - J. Med. Chem.
VL  - 63
IS  - 8
PB  - Amer Chemical Soc
PY  - 2020
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Glutathione peroxidase 4 (GPX4) is essential for cell membrane repair, inflammation suppression, and ferroptosis inhibition. GPX4 upregulation provides unique drug discovery opportunities for inflammation and ferroptosis-related diseases. However, rational design of protein activators is challenging. Until now, no compound has been reported to activate the enzyme activity of GPX4. Here, we identified a potential allosteric site in GPX4 and successfully found eight GPX4 activators using a novel computational strategy and experimental studies. Compound 1 from the virtual screen increased GPX4 activity, suppressed ferroptosis, reduced pro-inflammatory lipid mediator production, and inhibited NF-κB pathway activation. Further chemical synthesis and structure-activity relationship studies revealed seven more activators. The strongest compound, 1d4, increased GPX4 activity to 150% at 20 μM in the cell-free assay and 61 μM in cell extracts. Therefore, we demonstrated that GPX4 can be directly activated using chemical compounds to suppress ferroptosis and inflammation. Meanwhile, the discovery of GPX4 activators verified the possibility of rational design of allosteric activators.
AU  - Li, C.*
AU  - Deng, X.*
AU  - Zhang, W.*
AU  - Xie, X.*
AU  - Conrad, M.
AU  - Liu, Y.*
AU  - Friedmann Angeli, J.P.F.*
AU  - Lai, L.*
C1  - 55202
C2  - 46090
SP  - 266-275
TI  - Novel allosteric activators for ferroptosis regulator glutathione peroxidase 4.
JO  - J. Med. Chem.
VL  - 62
IS  - 1
PY  - 2019
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - The neutrophilic serine protease proteinase 3 (PR3) is involved in inflammation and immune response and thus appears as a therapeutic target for a variety of infectious and inflammatory diseases. Here we combined kinetic and molecular docking studies to increase the potency of peptidyl-diphenyl phosphonate PR3 inhibitors. Occupancy of the S1 subsite of PR3 by a nVal residue and of the S4-S5 subsites by a biotinylated Val residue as obtained in biotin-VYDnV(O-CH-4-Cl) enhanced the second-order inhibition constant k/[I] toward PR3 by more than 10 times ( k/[I] = 73000 ± 5000 M s) as compared to the best phosphonate PR3 inhibitor previously reported. This inhibitor shows no significant inhibitory activity toward human neutrophil elastase and resists proteolytic degradation in sputa from cystic fibrosis patients. It also inhibits macaque PR3 but not the PR3 from rodents and can thus be used for in vivo assays in a primate model of inflammation.
AU  - Guarino, C.*
AU  - Gruba, N.*
AU  - Grzywa, R.*
AU  - Dyguda-Kazimierowicz, E.*
AU  - Hamon, Y.*
AU  - Łȩgowska, M.*
AU  - Skoreński, M.*
AU  - Dallet-Choisy, S.*
AU  - Marchand-Adam, S.*
AU  - Kellenberger, C.*
AU  - Jenne, D.
AU  - Sieńczyk, M.*
AU  - Lesner, A.*
AU  - Gauthier, F.*
AU  - Korkmaz, B.*
C1  - 53276
C2  - 44465
CY  - Washington
SP  - 1858-1870
TI  - Exploiting the S4-S5 Specificity of Human Neutrophil Proteinase 3 to Improve the Potency of Peptidyl Di(chlorophenyl)-phosphonate Ester Inhibitors: A Kinetic and Molecular Modeling Analysis.
JO  - J. Med. Chem.
VL  - 61
IS  - 5
PB  - Amer Chemical Soc
PY  - 2018
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Celastrol is a natural pentacyclic triterpene used in traditional Chinese medicine with significant weight-lowering effects. Celastrol-administered mice at 100 mu g/kg decrease food consumption and body weight via a leptin-dependent mechanism, yet its molecular targets in this pathway remain elusive. Here, we demonstrate in vivo that celastrol-induced weight loss is largely mediated by the inhibition of leptin negative regulators protein tyrosine phosphatase (PTP) 1B (PTP1B) and T-cell PTP (TCPTP) in the arcuate nucleus (ARC) of the hypothalamus. We show in vitro that celastrol binds reversibly and inhibits noncompetitively PTP1B and TCPTP. NMR data map the binding site to an allosteric site in the catalytic domain that is in proximity of the active site. By using a panel of PTPs implicated in hypothalamic leptin signaling, we show that celastrol additionally inhibited PTEN and SHP2 but had no activity toward other phosphatases of the PTP family. These results suggest that PTP1B and TCPTP in the ARC are essential for celastrol's weight lowering effects in adult obese mice.
AU  - Kyriakou, E.
AU  - Schmidt, S.
AU  - Dodd, G.T.*
AU  - Pfuhlmann, K.
AU  - Simonds, S.E.*
AU  - Lenhart, D.
AU  - Geerlof, A.
AU  - Schriever, S.C.
AU  - de Angelis, M.
AU  - Schramm, K.-W.
AU  - Plettenburg, O.
AU  - Cowley, M.A.*
AU  - Tiganis, T.*
AU  - Tschöp, M.H.
AU  - Pfluger, P.T.
AU  - Sattler, M.
AU  - Messias, A.C.
C1  - 54937
C2  - 45971
CY  - 1155 16th St, Nw, Washington, Dc 20036 Usa
SP  - 11144-11157
TI  - Celastrol promotes weight loss in diet-induced obesity by inhibiting the protein tyrosine phosphatases PTP1B and TCPTP in the hypothalamus.
JO  - J. Med. Chem.
VL  - 61
IS  - 24
PB  - Amer Chemical Soc
PY  - 2018
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - 17β-HSD14 is a SDR enzyme, able to oxidize estradiol and 5-androstenediol using NAD(+). We determined the crystal structure of this human enzyme as the holo form and as ternary complexes with estrone and with the first potent, non-steroidal inhibitor. The structures reveal a conical, rather large and lipophilic binding site and are the starting point for structure-based inhibitor design. The two natural variants (S205 and T205) were characterized and adopt a similar structure.
AU  - Bertoletti, N.*
AU  - Braun, F.*
AU  - Lepage, M.*
AU  - Möller, G.
AU  - Adamski, J.
AU  - Heine, A.*
AU  - Klebe, G.*
AU  - Marchais-Oberwinkler, S.*
C1  - 48993
C2  - 41560
SP  - 6961-6967
TI  - New insights into human 17β-hydroxysteroid dehydrogenase type 14: First crystal structures in complex with a steroidal ligand and with a potent non-steroidal inhibitor.
JO  - J. Med. Chem.
VL  - 59
IS  - 14
PY  - 2016
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - 17β-HSD14 belongs to the SDR family and oxidizes the hydroxyl group at position 17 of estradiol and 5-androstenediol using NAD+ as cofactor. The goal of this study was to identify and optimize 17β-HSD14 nonsteroidal inhibitors as well as to disclose their structure-activity relationship. In a first screen, a library of 17β-HSD1 and 17β-HSD2 inhibitors, selected with respect to scaffold diversity, was tested for 17β-HSD14 inhibition. The most interesting hit was taken as starting point for chemical modification applying a ligand-based approach. The designed compounds were synthesized and tested for 17β-HSD14 inhibitory activity. The two best inhibitors identified in this study have a very high affinity to the enzyme with a Ki equal to 7 nM. The strong affinity of these inhibitors to the enzyme active site could be explained by crystallographic structure analysis, which highlighted the role of an extended H-bonding network in the stabilization process. The selectivity of the most potent compounds with respect to 17β-HSD1 and 17β-HSD2 is also addressed.
AU  - Braun, F.*
AU  - Bertoletti, N.*
AU  - Möller, G.
AU  - Adamski, J.
AU  - Steinmetzer, T.*
AU  - Salah, M.*
AU  - Abdelsamie, A.S.*
AU  - Van Koppen, C.J.*
AU  - Heine, A.*
AU  - Klebe, G.*
AU  - Marchais-Oberwinkler, S.*
C1  - 50170
C2  - 42054
CY  - Washington
SP  - 10719-10737
TI  - First structure-activity relationship of 17β-hydroxysteroid dehydrogenase type 14 nonsteroidal inhibitors and crystal structures in complex with the enzyme.
JO  - J. Med. Chem.
VL  - 59
IS  - 23
PB  - Amer Chemical Soc
PY  - 2016
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - U2AF homology motifs (UHMs) are atypical RNA recognition motif domains that mediate critical protein-protein interactions during the regulation of alternative pre-mRNA splicing and other processes. The recognition of UHM domains by UHM ligand motif (Ulm) peptide sequences plays important roles during early steps of spliceosome assembly. Splicing factor 45 kDa (SPF45) is an alternative splicing factor implicated in breast and lung cancers, and splicing regulation of apoptosis-linked pre-mRNAs by SPF45 was shown to depend on interactions between its UHM domain and Ulm motifs in constitutive splicing factors. We have developed cyclic peptide inhibitors that target UHM domains. By screening a focused library of linear and cyclic peptides and performing structure-activity relationship analysis, we designed cyclic peptides with 4-fold improved binding affinity for the SPF45 UHM domain compared to native Ulm ligands and 270-fold selectivity to discriminate UHM domains from alternative and constitutive splicing factors. These inhibitors are useful tools to modulate and dissect mechanisms of alternative splicing regulation.
AU  - Jagtap, P.K.
AU  - Garg, D.
AU  - Kapp, T.G.*
AU  - Will, C.L.*
AU  - Demmer, O.*
AU  - Lührmann, R.*
AU  - Kessler, H.*
AU  - Sattler, M.
C1  - 50098
C2  - 42107
CY  - Washington
SP  - 10190-10197
TI  - Rational design of cyclic peptide inhibitors of U2AF homology motif (UHM) domains to modulate pre-mRNA splicing.
JO  - J. Med. Chem.
VL  - 59
IS  - 22
PB  - Amer Chemical Soc
PY  - 2016
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Kinase inhibition is considered to be an important therapeutic target for LRRK2 mediated Parkinson's disease (PD). Many LRRK2 kinase inhibitors have been reported but have yet to be optimized in order to qualify as drug candidates for the treatment of the disease. In order to start a structure-function analysis of such inhibitors, we mutated the active site of Dictyostelium Roco4 kinase to resemble LRRK2. Here, we show saturation transfer difference (STD) NMR and the first cocrystal structures of two potent in vitro inhibitors, LRRK2-IN-1 and compound 19, with mutated Roco4. Our data demonstrate that this system can serve as an excellent tool for the structural characterization and optimization of LRRK2 inhibitors using X-ray crystallography and NMR spectroscopy.
AU  - Gilsbach, B.K.*
AU  - Messias, A.C.
AU  - Ito, G.*
AU  - Sattler, M.
AU  - Alessi, D.R.*
AU  - Wittinghofer, A.*
AU  - Kortholt, A.*
C1  - 44966
C2  - 37123
CY  - Washington
SP  - 3751-3756
TI  - Structural characterization of LRRK2 inhibitors.
JO  - J. Med. Chem.
VL  - 58
IS  - 9
PB  - Amer Chemical Soc
PY  - 2015
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Inhibition of 17β-HSD2 is an attractive mechanism for the treatment of osteoporosis. We report here the optimization of human 17β-HSD2 inhibitors in the 2,5-thiophene amide class by varying the size of the linker (n equals 0 and 2) between the amide moiety and the phenyl group. While none of the phenethylamides (n = 2) were active, most of the anilides (n = 0) turned out to moderately or strongly inhibit 17β-HSD2. The four most active compounds showed an IC(50) of around 60 nM and a very good selectivity toward 17β-HSD1, 17β-HSD4, 17β-HSD5, 11β-HSD1, 11β-HSD2 and the estrogen receptors α and β. The investigated compounds inhibited monkey 17β-HSD2 moderately, and one of them showed good inhibitory activity on mouse 17β-HSD2. SAR studies allowed a first characterization of the human 17β-HSD2 active site, which is predicted to be considerably larger than that of 17β-HSD1.
AU  - Marchais-Oberwinkler, S.*
AU  - Xu, K.*
AU  - Wetzel, M.*
AU  - Perspicace, E.*
AU  - Negri, M.*
AU  - Odermatt, A.*
AU  - Möller, G.
AU  - Adamski, J.
AU  - Hartmann, R.W.*
C1  - 11863
C2  - 30837
SP  - 167-181
TI  - Structural optimization of 2,5-tiophene amides as highly potent and selective 17β-hydroxysteroid dehydrogenase type 2 inhibitors for the treatment of osteoporosis.
JO  - J. Med. Chem.
VL  - 56
IS  - 1
PB  - American Chemical Society
PY  - 2013
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Estrogen deficiency in postmenopausal women or elderly men is often associated with the skeletal disease osteoporosis. The supplementation of estradiol (E2) in osteoporotic patients is known to prevent bone fracture but cannot be administered because of adverse effect. As 17β-hydroxysteroid dehydrogenase type 2 (17β-HSD2) oxidizes E2 to its inactive form estrone (E1) and has been found in osteoblastic cells, it is an attractive target for the treatment of osteoporosis. Twenty-one novel, naphthalene-derived compounds have been synthesized and evaluated for their 17β-HSD2 inhibition and their selectivity toward 17β-HSD1 and the estrogen receptors (ERs) α and β. Compound 19 turned out to be the most potent and selective inhibitor of 17β-HSD2 in cell-free assays and had a very good cellular activity in MDA-MB-231 cells, expressing naturally 17β-HSD2. It also showed marked inhibition of the E1-formation by the rat and mouse orthologous enzymes and strong inhibition of monkey 17β-HSD2. It is thus an appropriate candidate to be further evaluated in a disease-oriented model.
AU  - Wetzel, M.*
AU  - Marchais-Oberwinkler, S.*
AU  - Perspicace, E.*
AU  - Möller, G.
AU  - Adamski, J.
AU  - Hartmann, R.W.*
C1  - 6749
C2  - 29205
CY  - Washington, DC
SP  - 7547-7557
TI  - Introduction of an electron withdrawing group on the hydroxyphenylnaphthol scaffold improves the potency of 17β-hydroxysteroid dehydrogenase type 2 (17β-HSD2) inhibitors.
JO  - J. Med. Chem.
VL  - 54
IS  - 21
PB  - American Chemical Society
PY  - 2011
SN  - 0022-2623
ER  - 

TY  - JOUR
AU  - Garg, D.
AU  - Henrich, S.
AU  - Salo-Ahen, O.M.H.
AU  - Myllykallio, H.
AU  - Costi, M.P.
AU  - Wade, R.C.
C1  - 5133
C2  - 28180
SP  - 6539-6549
TI  - Novel approaches for targeting thymidylate synthase to overcome the resistance and toxicity of anticancer drugs.
JO  - J. Med. Chem.
VL  - 53
IS  - 18
PB  - Amer. Chemical Society
PY  - 2010
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - Porphycene photosensitizers bearing two or four methoxyethyl side chains were synthesized in nine steps from commercially available starting materials. Ether cleavage led to (hydroxyethyl)- and (bromoethyl)porphycenes that were converted to vinyl and benzo derivatives. Five of the side chain-functionalized porphycenes were biologically studied in comparison with two tetra-n-propylporphycenes. Porphycenes were incorporated in small unilamellar liposomes and incubated with cultivated SSK2 murine fibrosarcoma cells. Cellular uptake and phototoxicity 24 h after 5 J/cm2 laser light treatment were determined. The porphycenes tested were between 17 and 220 times more photodynamically active than the currently clinically used sensitizer Photofrin, although extinction coefficients of the porphycenes' irradiated bands are only approximately 10-fold higher. The LD50 concentration for SSK2 cells in the incubation medium was as low as (8.5 ± 2.8) × 10-9 M for tetrakis(methoxyethyl)porphycene. Two methoxy or hydroxy groups enhanced cellular uptake, three or four methoxy groups both enhanced and accelerated cellular uptake of tetraalkylporphycenes. Half-life times of the uptake processes varied between (0.14 ± 0.04) and (14 ± 4) h and cellular saturation levels between (1.2 ± 0.2) and (26 ± 3) pmol/105 cells. When individual uptake rates were accounted for, all porphycenes had a similar "cellular" phototoxicity, pointing toward a common mechanism of action. Evidence is presented for the assumption that cell membranes are the primary targets of the tested porphycenes and that membrane solubility may play a critical role in their photodynamic efficiency. The results show that nonionic polar side chain functionalities can strongly enhance cellular uptake and antitumor activity of lipophilic porphyrinoids and thus that the known lipophilicity/activity relationship can be reversed for very hydrophobic sensitizers.
AU  - Richert, C.*
AU  - Wessels, J.M.
AU  - Müller, M.D.*
AU  - Kisters, M.*
AU  - Benninghaus, T.*
AU  - Goetz, A.E.*
C1  - 40038
C2  - 37860
SP  - 2797-2807
TI  - Photodynamic antitumor agents: β-Methoxyethyl groups give access to functionalized porphycenes and enhance cellular uptake and activity.
JO  - J. Med. Chem.
VL  - 37
IS  - 17
PY  - 1994
SN  - 0022-2623
ER  - 

TY  - JOUR
AU  - Strobl, G.R.
AU  - Kruedener, S. von
AU  - Stöckigt, J.
AU  - Guengerich, F.P.
AU  - Wolff, T.
C1  - 20329
C2  - 13520
SP  - 1136-1145
TI  - Development of a Pharmacophore for Inhibition of Human Liver Cytochrome P-450 2D6: Molecular Modeling and Inhibition Studies.
JO  - J. Med. Chem.
VL  - 36
PY  - 1993
SN  - 0022-2623
ER  - 

TY  - JOUR
AB  - To gain insight into the specificity of cytochrome P-450 2D6 toward inhibitors, a preliminary pharmacophore model was built up using strong competitive inhibitors. Ajmalicine (1), the strongest inhibitor known (Ki = 3 nM) was selected as template because of its rigid structure. The preliminary pharmacophore model was validated by performing inhibition studies with derivatives of ajmalicine (1) and quinidine (9). Bufuralol (18) was chosen as substrate and the metabolite 1′-hydroxybufuralol (19) was separated by high performance liquid chromatography. All incubations were carried out using human liver microsomes after demonstration that the Ki values obtained with microsomes were in accordance with those obtained with a reconstituted monooxygenase system containing purified cytochrome P-450 2D6. Large differences of Ki values ranging between 0.005 and 100 μM were observed. Low-energy conformers of tested compounds were fit within the preliminary pharmacophore model. The analysis of steric and electronic properties of these compounds led to the definition of a final pharmacophore model. Characteristic properties are a positive charge on a nitrogen atom and a flat hydrophobic region, the plane of which is almost perpendicular to the N-H axis and maximally extends up to a distance of 7.5 Å from the nitrogen atom. Compounds with high inhibitory potency had additional functional groups with negative molecular electrostatic potential and hydrogen bond acceptor properties on the opposite side at respective distances of 4.8-5.5 Å and 6.6-7.5 Å from the nitrogen atom. The superposition of strong and weak inhibitors led to the definition of an excluded volume map. Compounds that required additional space were not inhibitors. This is apparently the first pharmacophore model for inhibitors of a cytochrome P-450 enzyme and offers the opportunity to classify compounds according to their potency of inhibition. Adverse drug interactions which occur when both substrates and inhibitors of cytochrome P-450 2D6 are applied may be predicted.
AU  - Strobl, G.R.
AU  - von Kruedener, S.*
AU  - Stöckigt, J.H.H.*
AU  - Guengerich, F.P.*
AU  - Wolff, T.
C1  - 40385
C2  - 0
SP  - 1136-1145
TI  - Development of a pharmacophore for inhibition of human liver cytochrome P-450 2D6: Molecular modeling and inhibition studies.
JO  - J. Med. Chem.
VL  - 36
IS  - 9
PY  - 1993
SN  - 0022-2623
ER  -