Discovery of dihydrothieno- and dihydrofuropyrimidines as potent pan Akt inhibitors

Bencsik, Josef R.; Xiao, Duan; Blake, James F.; Kallan, Nicholas C.; Mitchell, Ian S.; Spencer, Keith L.; Xu, Rui; Gloor, Susan L.; Martinson, Matthew; Risom, Tyler; Woessner, Richard; Dizon, Faith P.; Wu, Wen-I; Vigers, Guy; Brandhuber, Barbara J.; Skelton, Nicholas J.; Prior, Wei Wei; Murray, Lesley J.

doi:10.1016/j.bmcl.2010.09.112

Cited by 34 publications

(32 citation statements)

References 25 publications

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“…However, the baculovirus expression vector system (BEVS) can enable production of post-translationally-modified recombinant proteins. To date, a number of structural studies on the kinase domain of PKB1314151617 and of its complex structure with inhibitors18192021222324 have been performed using recombinant proteins derived from an Sf9 expression system. Unfortunately, the commercially available Sf9 expression systems using Autographa californica nucleopolyhedrovirus (AcNPV) BEVS have the major drawbacks of requiring large-scale culture of insect cell lines, a high-titer recombinant virus, and proficiency in time-consuming virus-handling techniques.…”

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confidence: 99%

Efficient and cost effective production of active-form human PKB using silkworm larvae

Maesaki

Satoh

Taoka

et al. 2014

Sci Rep

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Protein kinase B (PKB) also known as Akt is involved in many signal transduction pathways. As alterations of the PKB pathway are found in a number of human malignancies, PKB is considered an important drug target for cancer therapy. However, production of sufficient amounts of active PKB for biochemical and structural studies is very costly because of the necessity of using a higher organism expression system to obtain phosphorylated PKB. Here, we report efficient production of active PKBα using the BmNPV bacmid expression system with silkworm larvae. Following direct injection of bacmid DNA, recombinant PKBα protein was highly expressed in the fat bodies of larvae, and could be purified using a GST-tag and then cleaved. A final yield of approximately 1 mg PKBα/20 larvae was recorded. Kinase assays showed that the recombinant PKBα possessed high phosphorylation activity. We further confirmed phosphorylation on the activation loop by mass spectrometric analysis. Our results indicate that the silkworm expression system is of value for preparation of active-form PKBα with phosphorylation on the activation loop. This efficient production of the active protein will facilitate further biochemical and structural studies and stimulate subsequent drug development.

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confidence: 99%

Efficient and cost effective production of active-form human PKB using silkworm larvae

Maesaki

Satoh

Taoka

et al. 2014

Sci Rep

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“…The structure and bioactivity (IC 50 ) of the molecules were extracted from the literature . The bioactivity IC 50 (n m ) was determined with enzymatic assay by the previous work and converted to the logarithmic scale pIC 50 (n m ).…”

Section: Methodsmentioning

confidence: 99%

Selectivity Mechanism of ATP‐Competitive Inhibitors for PKB and PKA

Pang

Dong

et al. 2014

Chem Biol Drug Des

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Protein kinase B (PKB) acts as a central node on the PI3K kinase pathway. Constitutive activation and overexpression of PKB have been identified to involve in various cancers. However, protein kinase A (PKA) sharing high homology with PKB is essential for metabolic regulation. Therefore, specific targeting on PKB is crucial strategy in drug design and development for antitumor. Here, we had revealed the selectivity mechanism for PKB inhibitors with molecular dynamics simulation and 3D-QSAR methods. Selective inhibitors of PKB could form more hydrogen bonds and hydrophobic contacts with PKB than those with PKA. This could explain that selective inhibitor M128 is more potent to PKB than to PKA. Then, 3D-QSAR models were constructed for these selective inhibitors and evaluated by test set compounds. 3D-QSAR model comparison of PKB inhibitors and PKA inhibitors reveals possible methods to improve the selectivity of inhibitors. These models can be used to design new chemical entities and make quantitative prediction of the specific selective inhibitors before resorting to in vitro and in vivo experiment.

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“…In molecular docking simulations, Glide/XP docking protocols were applied for prediction of the topologies of compounds 3 and 8 in the active site of the target structure. 28…”

Section: Molecular Docking Studiesmentioning

confidence: 99%

Comprehensive Study on Thiadiazole-Based Anticancer Agents Inducing Cell Cycle Arrest and Apoptosis/Necrosis Through Suppression of Akt Activity in Lung Adenocarcinoma and Glioma Cells

Çiftçi

Sever

Altıntop

2019

tjps

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Amaç: Akt, antikanser ilaç keşfinde ve gelişiminde önemli bir hedef olarak düşünülmektedir ve bu nedenle Akt enzimini hedefleyen yeni potent antikanser ajanların keşfi için yoğun çaba harcanmıştır. Gereç ve Yöntemler: Tiyadiazollerin antikanser ilaç keşfindeki önemine bağlı olarak, bu çalışmada sekiz adet 1,3,4-tiyadiazol türevinin C6 sıçan glioma ve A549 insan akciğer adenokarsinoma hücre dizileri üzerindeki sitotoksik etkileri MTT deneyi kullanılarak araştırılmıştır. En etkili antikanser ajanların apoptoz, kaspaz-3 aktivasyonu, mitokondriyal membran potansiyeli, hücre döngüsü arresti üzerindeki etkileri BD FACSAria (I) akış sitometrisi ile belirlenmiştir. Akt aktivite, C6 ve A549 hücre dizilerinde ELISA kolorimetrik yöntem kullanılarak ölçülmüştür. 3 ve 8 no'lu bileşiklerin Akt enziminin (PDB kod: 3OW4) aktif bölgesindeki olası bağlanma biçimlerini araştırmak için Schrödinger's Maestro moleküler modelleme programı kullanılmıştır. Bulgular: N-(4-Klorofenil)-2-[(5-((4-nitrofenil)amino)-1,3,4-tiyadiazol-2-il)tiyo]asetamit (3) ve N-(6-nitrobenzotiyazol-2-il)-2-[(5-((4-nitrofenil) amino)-1,3,4-tiyadiazol-2-il)tiyo]asetamit (8), C6 hücre dizisinde apoptozu ve hücre döngüsü tutuklanmasını Akt aktivitesi inhibisyonu aracılığıyla Objectives: Akt is considered as an attractive target for anticancer drug discovery and development and therefore extensive efforts have been devoted to the discovery of new potent anticancer agents targeting Akt. Materials and Methods: Due to the importance of thiadiazoles for anticancer drug discovery, herein eight 1,3,4-thiadiazole derivatives were investigated for their cytotoxic effects on C6 rat glioma and A549 human lung adenocarcinoma cell lines using the MTT assay. The effects of the most promising anticancer agents on apoptosis, caspase-3 activation, mitochondrial membrane potential, and cell cycle arrest were determined on a BD FACSAria (I) flow cytometer. Akt activity was measured in the C6 and A549 cell lines using an ELISA colorimetric method. Schrödinger's Maestro molecular modeling package was used to explore the possible binding modes of compounds 3 and 8 in the active site of Akt enzyme (PDB code: 3OW4). Results: N-(4-Chlorophenyl)-2-[(5-((4-nitrophenyl)amino)-1,3,4-thiadiazol-2-yl)thio]acetamide (3) and N-(6-nitrobenzothiazol-2-yl)-2-[(5-((4nitrophenyl)amino)-1,3,4-thiadiazol-2-yl)thio]acetamide (8) induced apoptosis and cell cycle arrest in the C6 cell line through inhibition of Akt activity (92.36% and 86.52%, respectively). The docking results of compounds 3 and 8 indicated that π-π interactions, H bonds, and salt-bridge formation were responsible for the observed Akt inhibitory activity. Conclusion: According to in vitro and docking studies, compounds 3 and 8 stand out as promising antiglioma agents.

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Discovery of dihydrothieno- and dihydrofuropyrimidines as potent pan Akt inhibitors

Cited by 34 publications

References 25 publications

Efficient and cost effective production of active-form human PKB using silkworm larvae

Efficient and cost effective production of active-form human PKB using silkworm larvae

Selectivity Mechanism of ATP‐Competitive Inhibitors for PKB and PKA

Comprehensive Study on Thiadiazole-Based Anticancer Agents Inducing Cell Cycle Arrest and Apoptosis/Necrosis Through Suppression of Akt Activity in Lung Adenocarcinoma and Glioma Cells

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