109 results on '"Jaeki Min"'
Search Results
2. Discovery of novel, orally bioavailable, antileishmanial compounds using phenotypic screening.
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Diana Ortiz, W Armand Guiguemde, Jared T Hammill, Angela K Carrillo, Yizhe Chen, Michele Connelly, Kayla Stalheim, Carolyn Elya, Alex Johnson, Jaeki Min, Anang Shelat, David C Smithson, Lei Yang, Fangyi Zhu, R Kiplin Guy, and Scott M Landfear
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Arctic medicine. Tropical medicine ,RC955-962 ,Public aspects of medicine ,RA1-1270 - Abstract
Leishmaniasis is a parasitic infection that afflicts approximately 12 million people worldwide. There are several limitations to the approved drug therapies for leishmaniasis, including moderate to severe toxicity, growing drug resistance, and the need for extended dosing. Moreover, miltefosine is currently the only orally available drug therapy for this infection. We addressed the pressing need for new therapies by pursuing a two-step phenotypic screen to discover novel, potent, and orally bioavailable antileishmanials. First, we conducted a high-throughput screen (HTS) of roughly 600,000 small molecules for growth inhibition against the promastigote form of the parasite life cycle using the nucleic acid binding dye SYBR Green I. This screen identified approximately 2,700 compounds that inhibited growth by over 65% at a single point concentration of 10 μM. We next used this 2700 compound focused library to identify compounds that were highly potent against the disease-causing intra-macrophage amastigote form and exhibited limited toxicity toward the host macrophages. This two-step screening strategy uncovered nine unique chemical scaffolds within our collection, including two previously described antileishmanials. We further profiled two of the novel compounds for in vitro absorption, distribution, metabolism, excretion, and in vivo pharmacokinetics. Both compounds proved orally bioavailable, affording plasma exposures above the half-maximal effective concentration (EC50) concentration for at least 12 hours. Both compounds were efficacious when administered orally in a murine model of cutaneous leishmaniasis. One of the two compounds exerted potent activity against trypanosomes, which are kinetoplastid parasites related to Leishmania species. Therefore, this compound could help control multiple parasitic diseases. The promising pharmacokinetic profile and significant in vivo efficacy observed from our HTS hits highlight the utility of our two-step phenotypic screening strategy and strongly suggest that medicinal chemistry optimization of these newly identified scaffolds will lead to promising candidates for an orally available anti-parasitic drug.
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- 2017
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3. 8-Triazolylpurines: Towards Fluorescent Inhibitors of the MDM2/p53 Interaction.
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Mariell Pettersson, David Bliman, Jimmy Jacobsson, Jesper R Nilsson, Jaeki Min, Luigi Iconaru, R Kiplin Guy, Richard W Kriwacki, Joakim Andréasson, and Morten Grøtli
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Medicine ,Science - Abstract
Small molecule nonpeptidic mimics of α-helices are widely recognised as protein-protein interaction (PPIs) inhibitors. Protein-protein interactions mediate virtually all important regulatory pathways in a cell, and the ability to control and modulate PPIs is therefore of great significance to basic biology, where controlled disruption of protein networks is key to understanding network connectivity and function. We have designed and synthesised two series of 2,6,9-substituted 8-triazolylpurines as α-helix mimetics. The first series was designed based on low energy conformations but did not display any biological activity in a biochemical fluorescence polarisation assay targeting MDM2/p53. Although solution NMR conformation studies demonstrated that such molecules could mimic the topography of an α-helix, docking studies indicated that the same compounds were not optimal as inhibitors for the MDM2/p53 interaction. A new series of 8-triazolylpurines was designed based on a combination of docking studies and analysis of recently published inhibitors. The best compound displayed low micromolar inhibitory activity towards MDM2/p53 in a biochemical fluorescence polarisation assay. In order to evaluate the applicability of these compounds as biologically active and intrinsically fluorescent probes, their absorption/emission properties were measured. The compounds display fluorescent properties with quantum yields up to 50%.
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- 2015
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4. Identification of Selective Inhibitors of the Plasmodium falciparum Hexose Transporter PfHT by Screening Focused Libraries of Anti-Malarial Compounds.
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Diana Ortiz, W Armand Guiguemde, Alex Johnson, Carolyn Elya, Johanna Anderson, Julie Clark, Michele Connelly, Lei Yang, Jaeki Min, Yuko Sato, R Kiplin Guy, and Scott M Landfear
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Medicine ,Science - Abstract
Development of resistance against current antimalarial drugs necessitates the search for novel drugs that interact with different targets and have distinct mechanisms of action. Malaria parasites depend upon high levels of glucose uptake followed by inefficient metabolic utilization via the glycolytic pathway, and the Plasmodium falciparum hexose transporter PfHT, which mediates uptake of glucose, has thus been recognized as a promising drug target. This transporter is highly divergent from mammalian hexose transporters, and it appears to be a permease that is essential for parasite viability in intra-erythrocytic, mosquito, and liver stages of the parasite life cycle. An assay was developed that is appropriate for high throughput screening against PfHT based upon heterologous expression of PfHT in Leishmania mexicana parasites that are null mutants for their endogenous hexose transporters. Screening of two focused libraries of antimalarial compounds identified two such compounds that are high potency selective inhibitors of PfHT compared to human GLUT1. Additionally, 7 other compounds were identified that are lower potency and lower specificity PfHT inhibitors but might nonetheless serve as starting points for identification of analogs with more selective properties. These results further support the potential of PfHT as a novel drug target.
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- 2015
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5. Design, Synthesis and Evaluation of 2,5-Diketopiperazines as Inhibitors of the MDM2-p53 Interaction.
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Mariell Pettersson, Maria Quant, Jaeki Min, Luigi Iconaru, Richard W Kriwacki, M Brett Waddell, R Kiplin Guy, Kristina Luthman, and Morten Grøtli
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Medicine ,Science - Abstract
The transcription factor p53 is the main tumour suppressor in cells and many cancer types have p53 mutations resulting in a loss of its function. In tumours that retain wild-type p53 function, p53 activity is down-regulated by MDM2 (human murine double minute 2) via a direct protein-protein interaction. We have designed and synthesised two series of 2,5-diketopiperazines as inhibitors of the MDM2-p53 interaction. The first set was designed to directly mimic the α-helical region of the p53 peptide, containing key residues in the i, i+4 and i+7 positions of a natural α-helix. Conformational analysis indicated that 1,3,6-trisubstituted 2,5-diketopiperazines were able to place substituents in the same spatial orientation as an α-helix template. The key step of the synthesis involved the cyclisation of substituted dipeptides. The other set of tetrasubstituted 2,5-diketopiperazines were designed based on structure-based docking studies and the Ugi multicomponent reaction was used for the synthesis. This latter set comprised the most potent inhibitors which displayed micromolar IC50-values in a biochemical fluorescence polarisation assay.
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- 2015
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6. On the mechanism of action of SJ-172550 in inhibiting the interaction of MDM4 and p53.
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Michal Bista, David Smithson, Aleksandra Pecak, Gabriella Salinas, Katarzyna Pustelny, Jaeki Min, Artur Pirog, Kristin Finch, Michal Zdzalik, Brett Waddell, Benedykt Wladyka, Sylwia Kedracka-Krok, Michael A Dyer, Grzegorz Dubin, and R Kiplin Guy
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Medicine ,Science - Abstract
SJ-172550 (1) was previously discovered in a biochemical high throughput screen for inhibitors of the interaction of MDMX and p53 and characterized as a reversible inhibitor (J. Biol. Chem. 2010; 285:10786). Further study of the biochemical mode of action of 1 has shown that it acts through a complicated mechanism in which the compound forms a covalent but reversible complex with MDMX and locks MDMX into a conformation that is unable to bind p53. The relative stability of this complex is influenced by many factors including the reducing potential of the media, the presence of aggregates, and other factors that influence the conformational stability of the protein. This complex mechanism of action hinders the further development of compound 1 as a selective MDMX inhibitor.
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- 2012
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7. Protein–protein interfaces in molecular glue-induced ternary complexes: classification, characterization, and prediction
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Huan Rui, Kate S. Ashton, Jaeki Min, Connie Wang, and Patrick Ryan Potts
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Chemistry (miscellaneous) ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Molecular Biology ,Biochemistry - Abstract
This review surveys molecular glue-induced ternary complexes in the PDB and provides an overview of computational methods that can be utilized to predict them.
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- 2023
8. Combination of Ribociclib with BET-Bromodomain and PI3K/mTOR Inhibitors for Medulloblastoma Treatment In Vitro and In Vivo
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Barbara Jonchere, Justin Williams, Frederique Zindy, Jingjing Liu, Sarah Robinson, Dana M. Farmer, Jaeki Min, Lei Yang, Jennifer L. Stripay, Yingzhe Wang, Burgess B. Freeman, Jiyang Yu, Anang A. Shelat, Zoran Rankovic, and Martine F. Roussel
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Cancer Research ,Oncology ,Article - Abstract
Despite improvement in the treatment of medulloblastoma over the last years, numerous patients with MYC- and MYCN-driven tumors still fail current therapies. Medulloblastomas have an intact retinoblastoma protein RB, suggesting that CDK4/6 inhibition might represent a therapeutic strategy for which drug combination remains understudied. We conducted high-throughput drug combination screens in a Group3 (G3) medulloblastoma line using the CDK4/6 inhibitor (CDK4/6i) ribociclib at IC20, referred to as an anchor, and 87 oncology drugs approved by FDA or in clinical trials. Bromodomain and extra terminal (BET) and PI3K/mTOR inhibitors potentiated ribociclib inhibition of proliferation in an established cell line and freshly dissociated tumor cells from intracranial xenografts of G3 and Sonic hedgehog (SHH) medulloblastomas in vitro. A reverse combination screen using the BET inhibitor JQ1 as anchor, revealed CDK4/6i as the most potentiating drugs. In vivo, ribociclib showed single-agent activity in medulloblastoma models whereas JQ1 failed to show efficacy due to high clearance and insufficient free brain concentration. Despite in vitro synergy, combination of ribociclib with the PI3K/mTOR inhibitor paxalisib did not significantly improve the survival of G3 and SHH medulloblastoma-bearing mice compared with ribociclib alone. Molecular analysis of ribociclib and paxalisib-treated tumors revealed that E2F targets and PI3K/AKT/MTORC1 signaling genes were depleted, as expected. Importantly, in one untreated G3MB model HD-MB03, the PI3K/AKT/MTORC1 gene set was enriched in vitro compared with in vivo suggesting that the pathway displayed increased activity in vitro. Our data illustrate the difficulty in translating in vitro findings in vivo. See related article in Mol Cancer Ther (2022) 21(8):1306–1317.
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- 2022
9. Supplementary Table S4 from Combination of Ribociclib with BET-Bromodomain and PI3K/mTOR Inhibitors for Medulloblastoma Treatment In Vitro and In Vivo
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Martine F. Roussel, Zoran Rankovic, Anang A. Shelat, Jiyang Yu, Burgess B. Freeman, Yingzhe Wang, Jennifer L. Stripay, Lei Yang, Jaeki Min, Dana M. Farmer, Sarah Robinson, Jingjing Liu, Frederique Zindy, Justin Williams, and Barbara Jonchere
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Supplementary Table S4
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- 2023
10. Supplementary Methods, Figures and Tables from Combination of Ribociclib with BET-Bromodomain and PI3K/mTOR Inhibitors for Medulloblastoma Treatment In Vitro and In Vivo
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Martine F. Roussel, Zoran Rankovic, Anang A. Shelat, Jiyang Yu, Burgess B. Freeman, Yingzhe Wang, Jennifer L. Stripay, Lei Yang, Jaeki Min, Dana M. Farmer, Sarah Robinson, Jingjing Liu, Frederique Zindy, Justin Williams, and Barbara Jonchere
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Supplementary Methods, Figures S1-S9 and Tables S3, S5-S7
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- 2023
11. Ca2+-mediated mitochondrial inner membrane permeabilization induces cell death independently of Bax and Bak
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Giovanni Quarato, Fabien Llambi, Cliff S. Guy, Jaeki Min, Marisa Actis, Huan Sun, Shilpa Narina, Shondra M. Pruett-Miller, Junmin Peng, Zoran Rankovic, and Douglas R. Green
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Cell Biology ,Molecular Biology ,Article - Abstract
The ability of mitochondria to buffer a rapid rise in cytosolic Ca(2+) is a hallmark of proper cell homeostasis. Here, we employed m-3M3FBS, a putative phospholipase C (PLC) agonist, to explore the relationships between intracellular Ca(2+) imbalance, mitochondrial physiology, and cell death. m-3M3FBS induced a potent dose-dependent Ca(2+) release from the endoplasmic reticulum (ER), followed by a rise in intra-mitochondrial Ca(2+). When the latter exceeded the organelle buffering capacity, an abrupt mitochondrial inner membrane permeabilization (MIMP) occurred, releasing matrix contents into the cytosol. MIMP was followed by cell death that was independent of Bcl-2 family members and inhibitable by the intracellular Ca(2+) chelator BAPTA-AM. Cyclosporin A (CsA), capable of blocking the mitochondrial permeability transition (MPT), completely prevented cell death induced by m-3M3FBS. However, CsA acted upstream of mitochondria by preventing Ca(2+) release from ER stores. Therefore, loss of Ca(2+) intracellular balance and mitochondrial Ca(2+) overload followed by MIMP induced a cell death process that is distinct from Bcl-2 family-regulated mitochondrial outer membrane permeabilization (MOMP). Further, the inhibition of cell death by CsA or its analogues can be independent of effects on the MPT.
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- 2022
12. Development of Potent and Selective Janus Kinase 2/3 Directing PG–PROTACs
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Lisa J. Alcock, Yunchao Chang, Jamie A. Jarusiewicz, Marisa Actis, Stanley Nithianantham, Anand Mayasundari, Jaeki Min, Dylan Maxwell, Jeremy Hunt, Brandon Smart, Jun J. Yang, Gisele Nishiguchi, Marcus Fischer, Charles G. Mullighan, and Zoran Rankovic
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Organic Chemistry ,Drug Discovery ,Biochemistry - Abstract
[Image: see text] Aberrant activation of the JAK-STAT signaling pathway has been implicated in the pathogenesis of a range of hematological malignancies and autoimmune disorders. Here we describe the design, synthesis, and characterization of JAK2/3 PROTACs utilizing a phenyl glutarimide (PG) ligand as the cereblon (CRBN) recruiter. SJ10542 displayed high selectivity over GSPT1 and other members of the JAK family and potency in patient-derived ALL cells containing both JAK2 fusions and CRLF2 rearrangements.
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- 2022
13. Phenyl‐Glutarimides: Alternative Cereblon Binders for the Design of PROTACs
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Richard E. Lee, Jake Slavish, Brandon Young, Lei Yang, Sergio C. Chai, Marcus Fischer, Taosheng Chen, Zhenmei Li, Stephen W. White, Stanley Nithianantham, Jaeki Min, Gisele Nishiguchi, Seung Wook Yang, Patrick Ryan Potts, Marisa Actis, Shalandus H. Garrett, Jamie Jarusiewicz, Sourav Das, Martine F. Roussel, Barbara Jonchere, Anand Mayasundari, Yong Li, Mi-Kyung Yun, Xiang Fu, Fatemeh Keramatnia, Anang A. Shelat, and Zoran Rankovic
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BRD4 ,Ligand efficiency ,Hydrolysis ,Ubiquitin-Protein Ligases ,Cereblon ,Myeloid leukemia ,Glutarimide ,General Medicine ,General Chemistry ,Article ,Catalysis ,Bromodomain ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Proteolysis ,Humans ,Imide ,IC50 ,Piperidones ,Adaptor Proteins, Signal Transducing - Abstract
Targeting cereblon (CRBN) is currently one of the most frequently reported proteolysis-targeting chimera (PROTAC) approaches, owing to favorable drug-like properties of CRBN ligands, immunomodulatory imide drugs (IMiDs). However, IMiDs are known to be inherently unstable, readily undergoing hydrolysis in body fluids. Here we show that IMiDs and IMiD-based PROTACs rapidly hydrolyze in commonly utilized cell media, which significantly affects their cell efficacy. We designed novel CRBN binders, phenyl glutarimide (PG) analogues, and showed that they retained affinity for CRBN with high ligand efficiency (LE >0.48) and displayed improved chemical stability. Our efforts led to the discovery of PG PROTAC 4c (SJ995973), a uniquely potent degrader of bromodomain and extra-terminal (BET) proteins that inhibited the viability of human acute myeloid leukemia MV4–11 cells at low picomolar concentrations (IC(50) = 3 pM; BRD4 DC(50) = 0.87 nM). These findings strongly support the utility of PG derivatives in the design of CRBN-directed PROTACs.
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- 2021
14. Chemical scaffold recycling: Structure-guided conversion of an HIV integrase inhibitor into a potent influenza virus RNA-dependent RNA polymerase inhibitor designed to minimize resistance potential
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Peter J. Slavish, Maxime G. Cuypers, Mary Ashley Rimmer, Alireza Abdolvahabi, Trushar Jeevan, Gyanendra Kumar, Jamie A. Jarusiewicz, Sivaraja Vaithiyalingam, Jeremy C. Jones, John J. Bowling, Jeanine E. Price, Rebecca M. DuBois, Jaeki Min, Richard J. Webby, Zoran Rankovic, and Stephen W. White
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Pharmacology ,Organic Chemistry ,Drug Discovery ,General Medicine - Abstract
Influenza is one of the leading causes of disease-related mortalities worldwide. Several strategies have been implemented during the past decades to hinder the replication cycle of influenza viruses, all of which have resulted in the emergence of resistant virus strains. The most recent example is baloxavir marboxil, where a single mutation in the active site of the target endonuclease domain of the RNA-dependent-RNA polymerase renders the recent FDA approved compound ∼1000-fold less effective. Raltegravir is a first-in-class HIV inhibitor that shows modest activity to the endonuclease. Here, we have used structure-guided approaches to create rationally designed derivative molecules that efficiently engage the endonuclease active site. The design strategy was driven by our previously published structures of endonuclease-substrate complexes, which allowed us to target functionally conserved residues and reduce the likelihood of resistance mutations. We succeeded in developing low nanomolar equipotent inhibitors of both wild-type and baloxavir-resistant endonuclease. We also developed macrocyclic versions of these inhibitors that engage the active site in the same manner as their 'open' counterparts but with reduced affinity. Structural analyses provide clear avenues for how to increase the affinity of these cyclic compounds.
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- 2022
15. Preclinical Evaluation of Proteolytic Targeting of LCK as a Therapeutic Approach in T-cell Acute Lymphoblastic Leukemia
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Jianzhong Hu, Jamie Jarusiewicz, Guoqing Du, Gisele Nishiguchi, Satoshi Yoshimura, John C. Panetta, Zhenhua Li, Jaeki Min, Lei Yang, Divyabharathi Chepyala, Marisa Actis, Noemi Reyes, Brandon Smart, Ching-Hon Pui, David T. Teachey, Zoran Rankovic, and Jun J. Yang
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Lymphocyte Specific Protein Tyrosine Kinase p56(lck) ,Cell Line, Tumor ,T-Lymphocytes ,Ubiquitin-Protein Ligases ,Proteolysis ,Dasatinib ,Humans ,General Medicine ,Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ,Article - Abstract
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy and there is an unmet need for targeted therapies, especially for patients with relapsed disease. We have recently identified pre-T cell receptor (preTCR) and lymphocyte-specific protein tyrosine kinase (LCK) signaling as a common therapeutic vulnerability in T-ALL. LCK inhibitor dasatinib showed efficacy against T-ALL in preclinical studies and also in patients with T-ALL, however, this is transient in most cases. Leveraging the Proteolysis Targeting Chimera (PROTAC) approach, we developed a series of LCK degraders using dasatinib as an LCK ligand and phenyl-glutarimide as a cereblon-directing moiety. Our lead compound SJ11646 exhibited marked efficiency in cereblon-mediated LCK degradation in T-ALL cells. Relative to dasatinib, SJ11646 showed up to three orders of magnitude higher cytotoxicity in LCK-activated T-ALL cell lines and primary leukemia samples in vitro, with drastically prolonged suppression of LCK signaling. In vivo pharmacokinetic and pharmacodynamic profiling indicated a 630% increase in the duration of LCK suppression by SJ11646 over dasatinib in patient-derived xenograft models of T-ALL, which translated into its extended leukemia-free survival over dasatinib in vivo. Lastly, SJ11646 retained a high binding affinity to 51 human kinases, particularly ABL1, KIT, and DDR1, all of which are known drug targets in other cancers. Taken together, our dasatinib-based phenyl-glutarimide PROTACs are promising therapeutic agents in T-ALL and also valuable tools for developing degradation-based therapeutics for other cancers.
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- 2022
16. Identification of Potent, Selective, and Orally Bioavailable Small-Molecule GSPT1/2 Degraders from a Focused Library of Cereblon Modulators
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Zoran Rankovic, Barbara Jonchere, Junmin Peng, Martine F. Roussel, Anand Mayasundari, Sourav Das, Gisele Nishiguchi, Shalandus H. Garrett, Shondra Miller, Jeanine E. Price, Kiran Kodali, Fatemeh Keramatnia, Divyabharathi Chepyala, Lei Yang, James B Papizan, Jaeki Min, Marcus Fischer, P. Jake Slavish, Marisa Actis, Kevin McGowan, Charles G. Mullighan, Yong Li, Xiang Fu, Jamie Jarusiewicz, Brandon Young, and Yunchao Chang
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Ubiquitin-Protein Ligases ,Phenotypic screening ,Administration, Oral ,Computational biology ,Molecular Dynamics Simulation ,01 natural sciences ,Article ,Small Molecule Libraries ,Ikaros Transcription Factor ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,In vivo ,Cell Line, Tumor ,Drug Discovery ,Animals ,Humans ,Adaptor Proteins, Signal Transducing ,Retrospective Studies ,030304 developmental biology ,0303 health sciences ,Binding Sites ,Chemistry ,Cereblon ,Rational design ,Small molecule ,Thalidomide ,0104 chemical sciences ,Bioavailability ,010404 medicinal & biomolecular chemistry ,Proteolysis ,Molecular Medicine ,Identification (biology) ,Target protein ,Half-Life ,Peptide Termination Factors - Abstract
Whereas the PROTAC approach to target protein degradation greatly benefits from rational design, the discovery of small-molecule degraders relies mostly on phenotypic screening and retrospective target identification efforts. Here, we describe the design, synthesis, and screening of a large diverse library of thalidomide analogues against a panel of patient-derived leukemia and medulloblastoma cell lines. These efforts led to the discovery of potent and novel GSPT1/2 degraders displaying selectivity over classical IMiD neosubstrates, such as IKZF1/3, and high oral bioavailability in mice. Taken together, this study offers compound 6 (SJ6986) as a valuable chemical probe for studying the role of GSPT1/2 in vitro and in vivo, and it supports the utility of a diverse library of CRBN binders in the pursuit of targeting undruggable oncoproteins.
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- 2021
17. Target and tissue selectivity of PROTAC degraders
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Robert G. Guenette, Seung Wook Yang, Jaeki Min, Baikang Pei, and Patrick Ryan Potts
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Ubiquitin-Protein Ligases ,Proteolysis ,General Chemistry ,Ligands - Abstract
Targeted protein degradation (TPD) strategies have revolutionized how scientists tackle challenging protein targets deemed undruggable with traditional small molecule inhibitors. Many promising campaigns to inhibit proteins have failed due to factors surrounding inhibition selectivity and targeting of compounds to specific tissues and cell types. One of the major improvements that PROTAC (proteolysis targeting chimera) and molecular glue technology can exert is highly selective control of target inhibition. Multiple studies have shown that PROTACs can gain selectivity for their protein targets beyond that of their parent ligands
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- 2022
18. Development of BODIPY FL Thalidomide As a High-Affinity Fluorescent Probe for Cereblon in a Time-Resolved Fluorescence Resonance Energy Transfer Assay
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Jaeki Min, Wenwei Lin, Lei Yang, Jiuyu Liu, Taosheng Chen, Richard E. Lee, and Yongtao Li
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Boron Compounds ,Ubiquitin-Protein Ligases ,Proteolysis ,Biomedical Engineering ,Pharmaceutical Science ,Bioengineering ,Ligands ,Article ,chemistry.chemical_compound ,Fluorescence Resonance Energy Transfer ,otorhinolaryngologic diseases ,medicine ,Adaptor Proteins, Signal Transducing ,Fluorescent Dyes ,Pharmacology ,medicine.diagnostic_test ,biology ,Chemistry ,Cereblon ,Organic Chemistry ,Resonance (chemistry) ,Fluorescence ,Thalidomide ,Ubiquitin ligase ,Biophysics ,biology.protein ,Time-resolved spectroscopy ,BODIPY ,Biotechnology ,medicine.drug - Abstract
Ligands for cereblon, a component of a functional E3 ligase complex that targets proteins for proteolysis, are critical for developing molecular glues and proteolysis-targeting chimeras (PROTACs), which have therapeutic implications for various diseases. However, the lack of sensitivity of previously reported assays limits characterization of cereblon ligands. To address this shortcoming, we developed BODIPY FL Thalidomide (10) as a high-affinity fluorescent probe for the human cereblon protein, with a K(d) value of 3.6 nM. We then used BODIPY FL Thalidomide (10) to develop a cereblon time-resolved fluorescence resonance energy transfer (TR-FRET) binding assay. The IC(50) values of the cereblon ligand pomalidomide (8) were 6.4 nM in our cereblon TR-FRET binding assay, 264.8 nM in a previously reported Cy5-conjugated thalidomide (7)-mediated fluorescence polarization (FP) assay, and 1.2 μM in a previously reported Cy5-conjugated cereblon modulator (compound 7) (9)-mediated TR-FRET assay, indicating that our cereblon TR-FRET binding assay is 41- and 187-fold more sensitive than these two previously published assays. With our cereblon TR-FRET binding assay, we detected binding of cereblon ligands but not binding of bromodomain-containing protein 4 or von Hippel–Lindau ligands, thereby demonstrating its selectivity. Our cereblon TR-FRET binding assay was very stable and detected changes in phthalimide activity due to thalidomide isomerization. Therefore, the BODIPY FL Thalidomide (10)-mediated cereblon TR-FRET binding assay we designed is highly sensitive, selective, and stable and will aid the development and characterization of novel cereblon ligands.
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- 2020
19. Structural basis for substrate recognition and chemical inhibition of oncogenic MAGE ubiquitin ligases
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Elizabeth C. Griffith, Anand Mayasundari, Darcie J. Miller, Patrick Rodrigues, Zoran Rankovic, Jaeki Min, Patrick Ryan Potts, Clifford T. Gee, Richard E. Lee, Seung Wook Yang, Xin Huang, Wenwei Lin, Taosheng Chen, Lei Li, and Wei Li
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0301 basic medicine ,Carcinogenesis ,Amino Acid Motifs ,General Physics and Astronomy ,medicine.disease_cause ,law.invention ,Substrate Specificity ,0302 clinical medicine ,Ubiquitin ,law ,Neoplasms ,Protein Interaction Mapping ,lcsh:Science ,Substrate Interaction ,Multidisciplinary ,biology ,Chemistry ,Cell biology ,Neoplasm Proteins ,030220 oncology & carcinogenesis ,Aminoquinolines ,Protein Binding ,endocrine system ,Ubiquitin-Protein Ligases ,Science ,Drug development ,Antineoplastic Agents ,Proof of Concept Study ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Structure-Activity Relationship ,Antigen ,Protein Domains ,Antigens, Neoplasm ,medicine ,Structure–activity relationship ,Humans ,neoplasms ,mRNA Cleavage and Polyadenylation Factors ,HEK 293 cells ,Ubiquitination ,General Chemistry ,High-Throughput Screening Assays ,030104 developmental biology ,HEK293 Cells ,Ubiquitin ligases ,Mutagenesis ,biology.protein ,Suppressor ,lcsh:Q ,Degron ,Drug Screening Assays, Antitumor ,HeLa Cells - Abstract
Testis-restricted melanoma antigen (MAGE) proteins are frequently hijacked in cancer and play a critical role in tumorigenesis. MAGEs assemble with E3 ubiquitin ligases and function as substrate adaptors that direct the ubiquitination of novel targets, including key tumor suppressors. However, how MAGEs recognize their targets is unknown and has impeded the development of MAGE-directed therapeutics. Here, we report the structural basis for substrate recognition by MAGE ubiquitin ligases. Biochemical analysis of the degron motif recognized by MAGE-A11 and the crystal structure of MAGE-A11 bound to the PCF11 substrate uncovered a conserved substrate binding cleft (SBC) in MAGEs. Mutation of the SBC disrupted substrate recognition by MAGEs and blocked MAGE-A11 oncogenic activity. A chemical screen for inhibitors of MAGE-A11:substrate interaction identified 4-Aminoquinolines as potent inhibitors of MAGE-A11 that show selective cytotoxicity. These findings provide important insights into the large family of MAGE ubiquitin ligases and identify approaches for developing cancer-specific therapeutics., Testis-restricted melanoma antigen (MAGE) proteins function as substrate adapters for E3 ubiquitin ligases. Biochemical and structural analyses of MAGE-A11 provide insight into the substrate binding mode of MAGE proteins and enable discovery of potent, cytotoxic inhibitors of MAGE-A11:substrate interaction.
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- 2020
20. Ca
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Giovanni, Quarato, Fabien, Llambi, Cliff S, Guy, Jaeki, Min, Marisa, Actis, Huan, Sun, Shilpa, Narina, Shondra M, Pruett-Miller, Junmin, Peng, Zoran, Rankovic, and Douglas R, Green
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Editorial ,Mitochondrial Membranes ,Apoptosis ,Calcium ,Mitochondria ,bcl-2-Associated X Protein - Abstract
The ability of mitochondria to buffer a rapid rise in cytosolic Ca
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- 2021
21. Discovery of Novel Pyrazolo-pyridone DCN1 Inhibitors Controlling Cullin Neddylation
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Bhuvanesh Singh, Jonah Rector, Jaeki Min, Brenda A. Schulman, R. Kiplin Guy, Jared T. Hammill, Ho Shin Kim, Daniel C. Scott, and Yizhe Chen
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NEDD8 Protein ,Pyridones ,Glycine ,Cyclopentanes ,01 natural sciences ,NEDD8 ,Article ,Structure-Activity Relationship ,03 medical and health sciences ,Protein Domains ,Cell Line, Tumor ,Protein Interaction Mapping ,Drug Discovery ,Humans ,Structure–activity relationship ,Cell Proliferation ,030304 developmental biology ,0303 health sciences ,biology ,Chemistry ,Intracellular Signaling Peptides and Proteins ,Fibroblasts ,Cullin Proteins ,Amides ,Small molecule ,0104 chemical sciences ,Squamous carcinoma ,Cell biology ,010404 medicinal & biomolecular chemistry ,Pyrimidines ,Pevonedistat ,Drug Design ,Ubiquitin-Conjugating Enzymes ,Carcinoma, Squamous Cell ,biology.protein ,Pyrazoles ,Molecular Medicine ,CUL1 ,Neddylation ,Reactive Oxygen Species ,Cullin - Abstract
Chemical control of cullin neddylation is attracting increased attention based largely on the successes of the NEDD8-activating enzyme (E1) inhibitor Pevonedistat. Recently reported chemical probes enable selective and time-dependent inhibition of downstream members of the neddylation tri-enzymatic cascade including the co-E3, DCN1. In this work, we report the optimization of a novel class of small molecule inhibitors of the DCN1-UBE2M interaction. Rational X-ray co-structure enabled optimization afforded a 25-fold improvement in potency relative to the initial screening hit. The potency gains are largely attributed to additional hydrophobic interactions mimicking the N-terminal acetyl group that drives binding of UBE2M to DCN1. The compounds inhibit the protein-protein interaction, block NEDD8 transfer in biochemical assays, engage DCN1 in cells, and selectively reduce the steady-state neddylation of Cul1 and Cul3 in two squamous carcinoma cell lines harboring DCN1 amplification.
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- 2019
22. Mechanistic Insight on the Mode of Action of Colletoic Acid
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Gustavo Palacios, Ikbale El Ayachi, Darcie J. Miller, Adaris Rodriguez-Cortes, Jaeki Min, Fatima Rivas, Richard E. Lee, Taotao Ling, Walter H. Lang, Gustavo A. Miranda-Carboni, and Elizabeth C. Griffith
- Subjects
Chemistry ,Hep G2 Cells ,Crystallography, X-Ray ,Article ,Mitochondria ,Protein Structure, Tertiary ,Mice ,HEK293 Cells ,Action (philosophy) ,Computational chemistry ,3T3-L1 Cells ,11-beta-Hydroxysteroid Dehydrogenase Type 1 ,Drug Discovery ,Animals ,Humans ,Molecular Medicine ,Enzyme Inhibitors ,Sesquiterpenes ,hormones, hormone substitutes, and hormone antagonists - Abstract
The natural product colletoic acid (CA) is a selective inhibitor of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which primarily converts cortisone to the active glucocorticoid (GC) cortisol. Here, CA's mode of action and its potential as a chemical tool to study intracellular GC signaling in adipogenesis are disclosed. 11β-HSD1 biochemical studies of CA indicated that its functional groups at C-1, C-4, and C-9 were important for enzymatic activity; an X-ray crystal structure of 11β-HSD1 bound to CA at 2.6 Å resolution revealed the nature of those interactions, namely, a close-fitting and favorable interactions between the constrained CA spirocycle and the catalytic triad of 11β-HSD1. Structure-activity relationship studies culminated in the development of a superior CA analogue with improved target engagement. Furthermore, we demonstrate that CA selectively inhibits preadipocyte differentiation through 11β-HSD1 inhibition, suppressing other relevant key drivers of adipogenesis (i.e., PPARγ, PGC-1α), presumably by negatively modulating the glucocorticoid signaling pathway. The combined findings provide an in-depth evaluation of the mode of action of CA and its potential as a tool compound to study adipose tissue and its implications in metabolic syndrome.
- Published
- 2019
23. Abstract 423: Induced proximity and proteolytic targeting of LCK as a novel therapeutic approach in T-cell leukemia
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Jianzhong Hu, Jamie Jarusiewicz, Jaeki Min, Lei Yang, Divyabharathi Chepyala, Marisa Actis, Guoqing Du, Brandon Smart, Dylan Maxwell, Boer Xie, Satoshi Yoshimura, Junming Peng, David T. Teachey, Zoran Rankovic, and Jun J. Yang
- Subjects
Cancer Research ,Oncology - Abstract
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive cancer, with a particularly dismal prognosis in patients with relapsed diseases. Our group recently discovered LCK dependency as a therapeutic vulnerability in 33% of T-ALL and LCK inhibitor dasatinib exhibited strong efficacy in vitro and in vivo (Nat Cancer 2, 284, 2021). However, the transient LCK inhibition by dasatinib resulted in only partial responses, and novel agents are needed to suppress LCK signaling in T-ALL in a sustained fashion to achieve long-term efficacy. We synthesized a set of proteolytic targeting chimeras (PROTACs) that bring LCK to cereblon (CRBN) E3 ligase for ubiquitination and degradation. LCK-PROTACs showed extremely high anti-leukemia sensitivity, i.e., up to 1,561.3 fold increase over dasatinib in LCK-activated T-ALL cell line. Complete LCK degradation was induced by most LCK-PROTACs. Based on solubility, permeability, stability, and anti-leukemia effects in vitro, we prioritized SJ001011646 for further evaluations. SJ001011646 induced 50% LCK degradation at the concentration at ~0.001nM, with superior cytotoxic effects in patient-derived primary T-ALL samples compared to dasatinib. In addition, we confirmed the formation of ternary complex of LCK, SJ001011646, and CRBN, using the AlphaLISA assay. In a wash-out experiment, LCK-dependent T-ALL cells were exposed to SJ001011646 or dasatinib for 18 hours and then monitored for viability recovery. Both dasatinib and SJ001011646 led to dramatic growth inhibition within 2 days. While dasatinib-treated cells started to recover 96 hours after drug removal, cells treated with SJ001011646 failed to recover even after 240 hours. To systematically identify therapeutic targets of SJ001011646, we performed proteomic profiling of T-ALL cells before and after drug treatment in vitro. Overall, 126,670 unique peptides were identified and mapped to 10,158 proteins, of which LCK was most significantly reduced by PROTAC. In parallel, kinome-binding profiling confirmed LCK as a primary target of SJ001011646. Finally, we explored pharmacokinetic and pharmacodynamic properties of SJ001011646 in vivo, using patient-derived xenograft models of T-ALL. Given at the same dosage level as a single injection, SJ001011646 showed 367% increase in the duration of LCK suppression compared to dasatinib. Taken together, we described the development and preclinical evaluation of LCK-targeting PROTACs in T-ALL. Inducing proximity to CRBN and therefore degradation, these agents can produce sustained LCK suppression with much improved anti-leukemia efficacy than small molecule inhibitors, and are therefore promising novel therapeutic agents in T-ALL. Citation Format: Jianzhong Hu, Jamie Jarusiewicz, Jaeki Min, Lei Yang, Divyabharathi Chepyala, Marisa Actis, Guoqing Du, Brandon Smart, Dylan Maxwell, Boer Xie, Satoshi Yoshimura, Junming Peng, David T. Teachey, Zoran Rankovic, Jun J. Yang. Induced proximity and proteolytic targeting of LCK as a novel therapeutic approach in T-cell leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 423.
- Published
- 2022
24. BRAF inhibition protects against hearing loss in mice
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Huizhan Liu, Lauryn E. Caster, Marisa Zallocchi, Jian Zuo, Zhenhang Xu, Jaeki Min, Eva M. Holland, David Z.Z. He, Matthew A. Ingersoll, Duane G. Currier, Tal Teitz, Taosheng Chen, and Emma A. Malloy
- Subjects
MAPK/ERK pathway ,Proto-Oncogene Proteins B-raf ,Hearing loss ,medicine.medical_treatment ,Antineoplastic Agents ,Deafness ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Ototoxicity ,Hair Cells, Auditory ,otorhinolaryngologic diseases ,Medicine ,Animals ,Humans ,Health and Medicine ,Hearing Loss ,Protein Kinase Inhibitors ,Research Articles ,030304 developmental biology ,Cisplatin ,0303 health sciences ,Chemotherapy ,Multidisciplinary ,biology ,business.industry ,Kinase ,Cyclin-dependent kinase 2 ,SciAdv r-articles ,Dabrafenib ,medicine.disease ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,medicine.symptom ,business ,medicine.drug ,Research Article - Abstract
Dabrafenib, an FDA-approved oral anticancer drug, prevents cisplatin- and noise-induced hearing loss in animal models., Hearing loss caused by noise, aging, antibiotics, and chemotherapy affects 10% of the world population, yet there are no Food and Drug Administration (FDA)-approved drugs to prevent it. Here, we screened 162 small-molecule kinase-specific inhibitors for reduction of cisplatin toxicity in an inner ear cell line and identified dabrafenib (TAFINLAR), a BRAF kinase inhibitor FDA-approved for cancer treatment. Dabrafenib and six additional kinase inhibitors in the BRAF/MEK/ERK cellular pathway mitigated cisplatin-induced hair cell death in the cell line and mouse cochlear explants. In adult mice, oral delivery of dabrafenib repressed ERK phosphorylation in cochlear cells, and protected from cisplatin- and noise-induced hearing loss. Full protection was achieved in mice with co-treatment with oral AZD5438, a CDK2 kinase inhibitor. Our study explores a previously unidentified cellular pathway and molecular target BRAF kinase for otoprotection and may advance dabrafenib into clinics to benefit patients with cisplatin- and noise-induced ototoxicity.
- Published
- 2020
25. Degradation of Janus kinases in CRLF2-rearranged acute lymphoblastic leukemia
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Kathryn G. Roberts, Lisa J Alcock, Lauren Rowland, Divyabharathi Chepyala, Mark R. Litzow, Elisabeth Paietta, Jamie Jarusiewicz, Baranda S Hansen, Lei Yang, Yunchao Chang, Shondra M. Pruett-Miller, Randolph Larsen, Toshihiko Imamura, Aman Seth, Zoran Rankovic, Koshi Akahane, Dylan Maxwell, Jaeki Min, Shanshan Yu-Chen Bradford, Jun J. Yang, Marisa Actis, Chunxu Qu, Charles G. Mullighan, Stanley Nithianantham, Marcus Fischer, Anand Mayasundari, and Hiroaki Goto
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Models, Molecular ,Ruxolitinib ,Immunology ,Plenary Paper ,Context (language use) ,Protein degradation ,Biochemistry ,Mice, Inbred NOD ,Cell Line, Tumor ,Nitriles ,Drug Discovery ,medicine ,Animals ,Humans ,Receptors, Cytokine ,Protein Kinase Inhibitors ,Janus Kinases ,Chemistry ,Gene Expression Regulation, Leukemic ,Cereblon ,Cell Biology ,Hematology ,Janus Kinase 2 ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,medicine.disease ,Leukemia ,Pyrimidines ,Proteolysis ,Cancer research ,Intercellular Signaling Peptides and Proteins ,Pyrazoles ,Female ,Signal transduction ,Janus kinase ,Tyrosine kinase ,medicine.drug - Abstract
CRLF2-rearranged (CRLF2r) acute lymphoblastic leukemia (ALL) accounts for more than half of Philadelphia chromosome-like (Ph-like) ALL and is associated with a poor outcome in children and adults. Overexpression of CRLF2 results in activation of Janus kinase (JAK)-STAT and parallel signaling pathways in experimental models, but existing small molecule inhibitors of JAKs show variable and limited efficacy. Here, we evaluated the efficacy of proteolysis-targeting chimeras (PROTACs) directed against JAKs. Solving the structure of type I JAK inhibitors ruxolitinib and baricitinib bound to the JAK2 tyrosine kinase domain enabled the rational design and optimization of a series of cereblon (CRBN)-directed JAK PROTACs utilizing derivatives of JAK inhibitors, linkers, and CRBN-specific molecular glues. The resulting JAK PROTACs were evaluated for target degradation, and activity was tested in a panel of leukemia/lymphoma cell lines and xenograft models of kinase-driven ALL. Multiple PROTACs were developed that degraded JAKs and potently killed CRLF2r cell lines, the most active of which also degraded the known CRBN neosubstrate GSPT1 and suppressed proliferation of CRLF2r ALL in vivo, e.g. compound 7 (SJ988497). Although dual JAK/GSPT1-degrading PROTACs were the most potent, the development and evaluation of multiple PROTACs in an extended panel of xenografts identified a potent JAK2-degrading, GSPT1-sparing PROTAC that demonstrated efficacy in the majority of kinase-driven xenografts that were otherwise unresponsive to type I JAK inhibitors, e.g. compound 8 (SJ1008030). Together, these data show the potential of JAK-directed protein degradation as a therapeutic approach in JAK-STAT–driven ALL and highlight the interplay of JAK and GSPT1 degradation activity in this context.
- Published
- 2020
26. Development of Second-Generation CDK2 Inhibitors for the Prevention of Cisplatin-Induced Hearing Loss
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Robert A. Hazlitt, Lei Yang, Jie Fang, Tal Teitz, Justine D. Bonga, Taosheng Chen, Jian Zuo, Asli N. Goktug, Duane G. Currier, Zoran Rankovic, Shiyong Diao, Luigi I. Iconaru, and Jaeki Min
- Subjects
0301 basic medicine ,Hearing loss ,Drug Evaluation, Preclinical ,Antineoplastic Agents ,Mice, Inbred Strains ,Pharmacology ,Protective Agents ,Article ,Kenpaullone ,Structure-Activity Relationship ,03 medical and health sciences ,Organ Culture Techniques ,Therapeutic index ,Ototoxicity ,In vivo ,Drug Discovery ,medicine ,Animals ,Humans ,Hearing Loss ,Protein Kinase Inhibitors ,Cisplatin ,biology ,Chemistry ,Cyclin-Dependent Kinase 2 ,Cyclin-dependent kinase 2 ,Imidazoles ,medicine.disease ,Cochlea ,Pyrimidines ,030104 developmental biology ,biology.protein ,Molecular Medicine ,medicine.symptom ,Immortalised cell line ,medicine.drug - Abstract
There are currently no FDA-approved therapies to prevent the hearing loss associated with the usage of cisplatin in chemotherapeutic regimens. We recently demonstrated that the pharmacologic inhibition with kenpaullone or genetic deletion of CDK2 preserved hearing function in animal models treated with cisplatin, which suggests that CDK2 is a promising therapeutic target to prevent cisplatin-induced ototoxicity. In this study, we identified two lead compounds, AT7519 and AZD5438, from a focused library screen of 187 CDK2 inhibitors, performed in an immortalized cell line derived from neonatal mouse cochleae treated with cisplatin. Moreover, we screened 36 analogues of AT7519 and identified analogue 7, which exhibited an improved therapeutic index. When delivered locally, analogue 7 and AZD5438 both provided significant protection against cisplatin-induced ototoxicity in mice. Thus, we have identified two additional compounds that prevent cisplatin-induced ototoxicity in vivo and provided further evidence that CDK2 is a druggable target for treating cisplatin-induced ototoxicity.
- Published
- 2018
27. Piperidinyl Ureas Chemically Control Defective in Cullin Neddylation 1 (DCN1)-Mediated Cullin Neddylation
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Bhuvanesh Singh, Lei Yang, Brenda A. Schulman, Jared T. Hammill, R.K. Guy, Jaeki Min, Fangyi Zhu, Gloria Holbrook, Amy Matheny, Michele Connelly, and Daniel C. Scott
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Models, Molecular ,0301 basic medicine ,Lung Neoplasms ,NEDD8 Protein ,Molecular Conformation ,Antineoplastic Agents ,Crystallography, X-Ray ,NEDD8 ,Article ,Ligase inhibitor ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Proto-Oncogene Proteins ,Drug Discovery ,Humans ,Basal cell ,biology ,Chemistry ,Ligand binding assay ,Intracellular Signaling Peptides and Proteins ,Proteins ,Cullin Proteins ,High-Throughput Screening Assays ,Cell biology ,030104 developmental biology ,Cell culture ,030220 oncology & carcinogenesis ,Ubiquitin-Conjugating Enzymes ,Carcinoma, Squamous Cell ,biology.protein ,Molecular Medicine ,CUL1 ,Neddylation ,Drug Screening Assays, Antitumor ,Cullin ,Protein Binding ,Signal Transduction - Abstract
We previously discovered and validated a class of piperidinyl ureas that regulate defective in cullin neddylation 1 (DCN1)-dependent neddylation of cullins. Here, we report preliminary structure-activity relationship studies aimed at advancing our high-throughput screen hit into a tractable tool compound for dissecting the effects of acute DCN1-UBE2M inhibition on the NEDD8/cullin pathway. Structure-enabled optimization led to a 100-fold increase in biochemical potency and modestly increased solubility and permeability as compared to our initial hit. The optimized compounds inhibit the DCN1-UBE2M protein-protein interaction in our TR-FRET binding assay and inhibit cullin neddylation in our pulse-chase NEDD8 transfer assay. The optimized compounds bind to DCN1 and selectively reduce steady-state levels of neddylated CUL1 and CUL3 in a squamous cell carcinoma cell line. Ultimately, we anticipate that these studies will identify early lead compounds for clinical development for the treatment of lung squamous cell carcinomas and other cancers.
- Published
- 2018
28. Progress in the Development of Preventative Drugs for Cisplatin-Induced Hearing Loss
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Jaeki Min, Robert A. Hazlitt, and Jian Zuo
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0301 basic medicine ,Cisplatin ,medicine.medical_specialty ,Hearing loss ,Chemistry ,Extramural ,Severe hearing loss ,3. Good health ,Clinical trial ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Drug development ,Protective Agents ,030220 oncology & carcinogenesis ,Drug Discovery ,medicine ,Molecular Medicine ,Effective treatment ,medicine.symptom ,Intensive care medicine ,medicine.drug - Abstract
Cisplatin is a highly effective treatment for malignant cancers and has become a cornerstone in chemotherapeutic regimens. Unfortunately, its use in the clinic is often coupled with a high incidence of severe hearing loss. Over the past few decades, enormous effort has been put forth to find protective agents that selectively protect against the ototoxic side effects of cisplatin and do not interfere with its antitumoral activity. Many therapies have been successful in preclinical work, but only a few have shown any protection in the clinic, and none have been approved by the FDA. This review summarizes the clinical and preclinical studies of the most effective small-molecule candidates currently in clinical trials, while also detailing their molecular mechanisms of action, to gain insight for future drug development in the field.
- Published
- 2018
29. AZD5438-PROTAC: A selective CDK2 degrader that protects against cisplatin- and noise-induced hearing loss
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Sarath Vijayakumar, Zoran Rankovic, Jaeki Min, Marisa Zallocchi, Santanu Hati, Yuju Li, Jian Zuo, Sándor Lovas, and Robert A. Hazlitt
- Subjects
Proteolysis ,Excitotoxicity ,Antineoplastic Agents ,Molecular Dynamics Simulation ,Protective Agents ,medicine.disease_cause ,Article ,Cell Line ,Structure-Activity Relationship ,Ototoxicity ,Drug Discovery ,medicine ,Animals ,Humans ,Protein Kinase Inhibitors ,Zebrafish ,Pharmacology ,Cisplatin ,Dose-Response Relationship, Drug ,Molecular Structure ,medicine.diagnostic_test ,biology ,Chemistry ,Kinase ,Cyclin-Dependent Kinase 2 ,Organic Chemistry ,Cyclin-dependent kinase 2 ,Imidazoles ,General Medicine ,medicine.disease ,Small molecule ,Cell biology ,Pyrimidines ,Hearing Loss, Noise-Induced ,biology.protein ,Noise-induced hearing loss ,medicine.drug - Abstract
Cyclin-dependent kinase 2 (CDK2) is a potential therapeutic target for the treatment of hearing loss and cancer. Previously, we identified AZD5438 and AT7519-7 as potent inhibitors of CDK2, however, they also targeted additional kinases, leading to unwanted toxicities. Proteolysis Targeting Chimeras (PROTACs) are a new promising class of small molecules that can effectively direct specific proteins to proteasomal degradation. Herein we report the design, synthesis, and characterization of PROTACs of AT7519-7 and AZD5438 and the identification of PROTAC-8, an AZD5438-PROTAC, that exhibits selective, partial CDK2 degradation. Furthermore, PROTAC-8 protects against cisplatin ototoxicity and kainic acid excitotoxicity in zebrafish. Molecular dynamics simulations reveal the structural requirements for CDK2 degradation. Together, PROTAC-8 is among the first-in-class PROTACs with in vivo therapeutic activities and represents a new lead compound that can be further developed for better efficacy and selectivity for CDK2 degradation against hearing loss and cancer.
- Published
- 2021
30. Abstract LBA002: Targeting GSPT1 by a novel cereblon E3 ligase modulator for the treatment of Acute Lymphoblastic Leukemia
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Fatemeh Keramatnia, Yunchao Chang, Gisele Nishiguchi, Jaeki Min, Charles Mullighan, Marcus Fischer, and Zoran Rankovic
- Subjects
Cancer Research ,Oncology - Abstract
Acute Lymphoblastic Leukemia (ALL), the most common childhood cancer and the second most common acute leukemia in adults, arises from clonal expansion of undifferentiated lymphoid precursor cells in bone marrow. Although the majority of childhood ALL cases harbor clonal genetic alterations of transcription factor (TF) gene mutations or rearrangements, TF alterations remain difficult therapeutic targets. Small molecule induced protein degradation is a novel strategy that can be applied to currently undruggable targets such as TF and fusion oncoproteins. In this paradigm, small molecule degraders (either PROTAC or Molecular Glue (MG)) redirect the cell’s endogenous ubiquitin proteasome system and induce ubiquitination of the target protein or non-native substrate of an E3 ligase (neosubstrate) and its subsequent proteasomal degradation. Recently, the CRBN E3 ligase modulator, CC90009 was reported to show potent antitumor activity in acute myeloid leukemia, leading to the identification of GSPT1 (G1 to S phase transition factor) as a CRBN neosubstrate. These findings suggest the potential of MGs to target unanticipated vulnerabilities in different malignancies. Here, using a structurally diverse and unique set of MGs (Molecular Glue Library (MGL)) with confirmed CRBN binding affinity, we sought to identify novel CRBN modulators through phenotypic and proteomic methods. Our screening of the MGL in a panel of representative acute leukemia cell lines, including the CRLF2-rearranged ALL cell line MHH-CALL-4 identified several active MGs with EC50 Citation Format: Fatemeh Keramatnia, Yunchao Chang, Gisele Nishiguchi, Jaeki Min, Charles Mullighan, Marcus Fischer, Zoran Rankovic, Fatemeh Keramatnia. Targeting GSPT1 by a novel cereblon E3 ligase modulator for the treatment of Acute Lymphoblastic Leukemia [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2021 Oct 7-10. Philadelphia (PA): AACR; Mol Cancer Ther 2021;20(12 Suppl):Abstract nr LBA002.
- Published
- 2021
31. Development of Proteolytic Targeting Chimeras to Target Lck in T-Cell Acute Lymphoblastic Leukemia
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Jianzhong Hu, Jaeki Min, Marisa Actis, Satoshi Yoshimura, Divyabharathi Chepyala, Zoran Rankovic, Jamie Jarusiewicz, Jun J. Yang, Dylan Maxwell, David T. Teachey, Lauren Rowland, Guoqing Du, Junmin Peng, Boer Xie, Lei Yang, and Brandon Smart
- Subjects
medicine.anatomical_structure ,Lymphoblastic Leukemia ,T cell ,Immunology ,Cancer research ,medicine ,Cell Biology ,Hematology ,Biology ,Biochemistry - Abstract
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy for which novel therapies are much needed especially in patients with relapsed diseases. By combining large-scale ex vivo pharmacotype profiling with network-based systems biology analyses, our group recently identified LCK dependency as a therapeutic vulnerability in 44% of T-ALL in children (Nat Cancer 2, 284-299, 2021). LCK inhibitors such as dasatinib exhibit striking anti-leukemia effects in this T-ALL subset. However, the transient LCK inhibition by dasatinib only resulted in incomplete response to monotherapy unless the drug was delivered continuously at a high level. Therefore, it is imperative to develop novel agents that produce sustained suppression of LCK signaling in T-ALL. To this end, we synthesized a set of proteolytic targeting chimeras (PROTACs) that target LCK and cereblon (CRBN) E3 ligase. These PROTACs bind and recruit LCK to CRBN E3 ligase, rendering LCK susceptible to ubiquitination and ultimately proteasomal degradation. Cell viability assay was performed in an LCK-dependent T-ALL cell line KOPT-K1 to determine its sensitivity to this panel of PROTACs. PROTACs showed up to 6.9-fold improvement in cytotoxicity relative to dasatinib. To validate PROTAC-induced LCK degradation, KOPT-K1 cells were treated with PROTACs or dasatinib at 100 nM for 24 hours and LCK protein was quantified by western blotting. LCK degradation occurred rapidly in the presence of PROTAC agents whereas dasatinib treatment did not affect LCK abundance. Furthermore, PROTAC-induced apoptosis of T-ALL cells was abolished by lenalidomide, a CRBN binder, suggesting that their cytotoxic effects were mediated by CRBN-driven LCK degradation. We also validated these findings in patient-derived T-ALL samples. In addition, we determined solubility, permeability, and stability of these PROTACs in vitro. Based on anti-leukemia effects as well as physical chemical properties, we prioritized PROTACs SJ001011646 and SJ001011447 as our top candidates for further evaluations. We hypothesized that the catalytic protein degradation by PROTACs will produce a more sustained suppression of the LCK signaling compared to transient LCK inhibition by dasatinib. To test this, we performed a wash-out assay comparing anti-leukemic effects of top PROTACs with dasatinib. KOPT-K1 cells were treated with vehicle, dasatinib, SJ001011646 and SJ001011447, respectively at 100 nM for 18 hours. Cells were then washed and placed in drug-free culture with viability monitored daily. The vehicle treated cells exhibited an exponential growth while drug treated groups showed dramatic growth inhibition within 2 days. Notably, dasatinib-treated cells continued to undergo apoptosis for 96 hours after drug removal before started to recover. By contrast, SJ001011447 treatment repressed cell growth for 144 hours post wash-out. Most impressively, there was no evidence of growth recovery in cells treated SJ001011646 even 240 hours after drug removal. In addition, we confirmed the formation of ternary complex of LCK, PROTAC, and E3 ligase, using the AlphaLISA assay. To systematically identify therapeutic targets of PROTACs, we performed proteomic profiling of KOPT-K1 cells before and after drug treatment in vitro. Cells were treated with vehicle, SJ001011447 or SJ001011646 at 100 nM for 24 hours before harvested for Tandem Mass Tag-based proteomic profiling. Overall, 126,670 unique peptides were identified and mapped to 10,158 proteins, of which 34 and 35 were significantly changed by SJ001011447 and SJ001011646, respectively (p2 or In conclusion, we developed LCK-targeting PROTACs with potent anti-leukemia effects. Highly effective in degrading LCK protein, these agents produced sustained LCK suppression superior to small molecule inhibitors, pointing to novel strategies to therapeutically target LCK in T-ALL. Disclosures No relevant conflicts of interest to declare.
- Published
- 2021
32. Development of Potent and Selective Janus Kinase 2/3 Directing PG-PROTACs.
- Author
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Alcock, Lisa J., Yunchao Chang, Jarusiewicz, Jamie A., Actis, Marisa, Nithianantham, Stanley, Mayasundari, Anand, Jaeki Min, Maxwell, Dylan, Hunt, Jeremy, Smart, Brandon, Yang, Jun J., Nishiguchi, Gisele, Fischer, Marcus, Mullighan, Charles G., and Rankovic, Zoran
- Published
- 2022
- Full Text
- View/download PDF
33. Ventromorphins: A New Class of Small Molecule Activators of the Canonical BMP Signaling Pathway
- Author
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David B. Finkelstein, Karen Vrijens, Taosheng Chen, Jose Grenet, Jaeki Min, Wilson K. Clements, R. Kiplin Guy, Anang A. Shelat, Wenwei Lin, Martine F. Roussel, Dana Farmer, and Jamie R. Genthe
- Subjects
0301 basic medicine ,animal structures ,Cellular differentiation ,Smad Proteins ,Biology ,Bone morphogenetic protein ,Biochemistry ,Article ,Cell Line ,Myoblasts ,Small Molecule Libraries ,Mice ,03 medical and health sciences ,Chalcones ,Osteogenesis ,Cell Line, Tumor ,Drug Discovery ,Animals ,Humans ,BMP signaling pathway ,Zebrafish ,Osteoblasts ,Activator (genetics) ,Gene Expression Profiling ,Cell Differentiation ,General Medicine ,biology.organism_classification ,Molecular biology ,BMPR2 ,030104 developmental biology ,Gene Expression Regulation ,Bone Morphogenetic Proteins ,embryonic structures ,Molecular Medicine ,Signal transduction ,C2C12 ,Signal Transduction - Abstract
Here we describe three new small-molecule activators of BMP signaling found by high throughput screening of a library of ~600,000 small molecules. Using a cell-based luciferase assay in the BMP4-responsive human cervical carcinoma clonal cell line, C33A-2D2, we identified three compounds with similar chemotypes that each ventralize zebrafish embryos and stimulate increased expression of the BMP target genes, bmp2b and szl. Because these compounds ventralize zebrafish embryos, we have termed them “ventromorphins.” As expected for BMP pathway activators, they induce the differentiation of C2C12 myoblasts to osteoblasts. Affymetrix RNA analysis confirmed the differentiation results and showed that ventromorphins treatment elicits a response similar to BMP4 treatment. Unlike Isoliquiritigenin (SJ000286237), a flavone that maximally activates the pathway after 24 hours of treatment, all three ventromorphins induced SMAD1/5/8 phosphorylation within 30 minutes of treatment and achieved peak activity within 1 hour, indicating that their responses are consistent with directly activated BMP signaling.
- Published
- 2017
34. Performance of a docking/molecular dynamics protocol for virtual screening of nutlin-class inhibitors of Mdmx
- Author
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Kristin Finch, Jaeki Min, R. Kiplin Guy, Nagakumar Bharatham, Donald Bashford, Michael A. Dyer, and Anand Mayasundari
- Subjects
0301 basic medicine ,MDMX ,Antineoplastic Agents ,Cell Cycle Proteins ,Computational biology ,Molecular Dynamics Simulation ,01 natural sciences ,Molecular Docking Simulation ,Article ,Piperazines ,03 medical and health sciences ,chemistry.chemical_compound ,Computational chemistry ,Proto-Oncogene Proteins ,Materials Chemistry ,Humans ,Physical and Theoretical Chemistry ,Spectroscopy ,Virtual screening ,Binding Sites ,Imidazoles ,Nuclear Proteins ,Nutlin ,AutoDock ,Computer Graphics and Computer-Aided Design ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,Improved performance ,030104 developmental biology ,chemistry ,Docking (molecular) ,Drug Screening Assays, Antitumor ,Hydrophobic and Hydrophilic Interactions ,Protein Binding - Abstract
A virtual screening protocol involving docking and molecular dynamics has been tested against the results of fluorescence polarization assays testing the potency of a series of compounds of the nutlin class for inhibition of the interaction between p53 and Mdmx, an interaction identified as a driver of certain cancers. The protocol uses a standard docking method (AutoDock) with a cutoff based on the AutoDock score (ADscore), followed by molecular dynamics simulation with a cutoff based on root-mean-square-deviation (RMSD) from the docked pose. An analysis of the experimental and computational results shows modest performance of ADscore alone, but dramatically improved performance when RMSD is also used.
- Published
- 2017
35. Optimization of a Novel Series of Ataxia-Telangiectasia Mutated Kinase Inhibitors as Potential Radiosensitizing Agents
- Author
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Clementine Feau, Brandon Young, Yizhe Chen, Kexiao Guo, Gloria Holbrook, Jaeki Min, Michele Connelly, R. Kiplin Guy, Andrew Lemoff, Fangyi Zhu, Praveen Kumar Suryadevara, and Michael B. Kastan
- Subjects
0301 basic medicine ,Radiation-Sensitizing Agents ,Ataxia Telangiectasia Mutated Proteins ,In Vitro Techniques ,Pharmacology ,Substrate Specificity ,Colony-Forming Units Assay ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Drug Delivery Systems ,Pharmacokinetics ,Drug Discovery ,Quinazoline ,Animals ,Humans ,Structure–activity relationship ,Potency ,Kinase ,Chemistry ,Mice, Inbred C57BL ,030104 developmental biology ,Biochemistry ,Drug Design ,MCF-7 Cells ,Microsomes, Liver ,Quinazolines ,Molecular Medicine ,Female ,Radiosensitizing Agent ,Selectivity - Abstract
We previously reported a novel inhibitor of the ataxia-telangiectasia mutated (ATM) kinase, which is a target for novel radiosensitizing drugs. While our initial lead, compound 4, was relatively potent and nontoxic, it exhibited poor stability to oxidative metabolism and relatively poor selectivity against other kinases. The current study focused on balancing potency and selectivity with metabolic stability through structural modification to the metabolized site on the quinazoline core. We performed extensive structure-activity and structure-property relationship studies on this quinazoline ATM kinase inhibitor in order to identify structural variants with enhanced selectivity and metabolic stability. We show that, while the C-7-methoxy group is essential for potency, replacing the C-6-methoxy group considerably improves metabolic stability without affecting potency. Promising analogues 20, 27g, and 27n were selected based on in vitro pharmacology and evaluated in murine pharmacokinetic and tolerability studies. Compound 27g possessed significantly improve pharmacokinetics relative to that of 4. Compound 27g was also significantly more selective against other kinases than 4. Therefore, 27g is a good candidate for further development as a potential radiosensitizer.
- Published
- 2016
36. Identification of Toll-like receptor signaling inhibitors based on selective activation of hierarchically acting signaling proteins
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Hans Häcker, Julie A. Pollock, Wenwei Lin, Taosheng Chen, Vanessa Redecke, Yizhe Chen, R. Kiplin Guy, Sirish K. Ippagunta, Naina Sharma, Kazuki Tawaratsumida, Jaeki Min, Anthony A. High, and John A. Katzenellenbogen
- Subjects
0301 basic medicine ,TIRAP ,Phenotypic screening ,Quantitative proteomics ,Drug Evaluation, Preclinical ,Biochemistry ,Article ,Small Molecule Libraries ,Inhibitory Concentration 50 ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Piperidines ,Animals ,Humans ,Protein Interaction Maps ,Receptor ,Molecular Biology ,Cell Proliferation ,Toll-like receptor ,Dose-Response Relationship, Drug ,biology ,Toll-Like Receptors ,HEK 293 cells ,Signal transducing adaptor protein ,Cell Biology ,Ubiquitin ligase ,Cell biology ,HEK293 Cells ,RAW 264.7 Cells ,030104 developmental biology ,030220 oncology & carcinogenesis ,biology.protein ,Pyrazoles ,Protein Binding ,Signal Transduction - Abstract
Toll-like receptors (TLRs) recognize various pathogen- and host tissue-derived molecules and initiate inflammatory immune responses. Exaggerated or prolonged TLR activation, however, can promote etiologically diverse diseases, such as bacterial sepsis, metabolic and autoimmune diseases, or stroke. Despite the apparent medical need, no small molecule drugs against TLR-pathways are clinically available. This may be because of the complex signaling mechanisms of TLRs, which are governed by a series of protein interactions initiated by Toll-interleukine-1 receptor (TIR)-domains found in TLRs and the cytoplasmic adapter proteins TIRAP and MyD88. Activation/oligomerization of TLRs and consecutive oligomerization of MyD88 and/or TIRAP leads to the recruitment of IRAK family members and the ubiquitin ligase TRAF6. Here, we developed a phenotypic drug screening system based on inducible dimerization of TIRAP, MyD88 and TRAF6, that ranked hits according to the position of action in the hierarchy of the TLR signaling cascade. From a bioactive compound library, we identified methyl-piperidino-pyrazole (MPP) as a TLR-specific inhibitor. Structure-activity relationship analysis, quantitative proteomics, protein interaction assays, and cellular thermal shift assays strongly suggest that MPP targets the TIR domain of MyD88. Chemical evolution of the original MPP-scaffold generated compounds with selectivity for distinct TLRs that interfered with specific TIR interactions. Administration of a MPP-analog to mice protected against LPS-mediated TNFα release. These results validate this phenotypic screening approach and implicate the MPP scaffold as starting point for anti-inflammatory drug development.
- Published
- 2018
37. PDTM-02. TARGETING THE RB PATHWAY IN MEDULLOBLASTOMA
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Jaeki Min, Zoran Rankovic, Burgess B. Freeman, Allison Pribnow, Martine F. Roussel, Anang A. Shelat, Jennifer L. Stripay, Frederique Zindy, and Barbara Jonchere
- Subjects
Medulloblastoma ,Cancer Research ,Oncology ,Pediatric Tumors ,business.industry ,Cancer research ,medicine ,Neurology (clinical) ,medicine.disease ,business - Abstract
Medulloblastoma (MB), the most common malignant pediatric brain tumor, is classified into four major molecularly and histopathologically distinct subgroups, among which MYC-driven Group 3 MBs confer a poor prognosis. The Cyclin D/CDK4/CDK6/RB pathway is frequently deregulated in MB leading to uncontrolled cell proliferation, but tumors express an intact RB protein (Northcott et al., Nature, 2017). Therefore, CDK4/6 inhibitor drugs offer a possible therapeutic approach to treat MBs. Because single agent therapy ultimately leads to drug resistance, we initiated in vitro combination drug screens to identify drug classes potentiating CDK4/6 inhibitors. We used Group 3 MB patient-derived orthotopic xenografts (PDOXs), a human cell line (HDMB03), and freshly dissociated tumor cells propagated only in the mouse brain. The drug screen included 90 compounds comprising targeted and cytotoxic drugs that are FDA approved or under active clinical investigation. Using a bioluminescence-based assay that measures ATP consumption (CellTiter-Glo) to evaluate the number of viable cells, these 90 compounds were screened in combination with a fixed concentration of ribociclib, one of the three FDA approved CDK4/6 inhibitors. The primary screen, carried out in HDMB03 cells, revealed several drugs with additive or synergistic potential when combined with ribociclib, including BET inhibitors, MEK inhibitors, PI3K/mTOR inhibitors and gemcitabine. We are currently evaluating the combination of brain penetrant compounds in Group 3 MB PDOXs. The identification of potent drug combinations should provide new therapeutic options for the treatment of Group 3 MB, one of the most difficult to treat.
- Published
- 2019
38. Discovery of an Orally Bioavailable Inhibitor of Defective in Cullin Neddylation 1 (DCN1)-Mediated Cullin Neddylation
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Bhuvanesh Singh, Yizhe Chen, R. Kiplin Guy, Brenda A. Schulman, Daniel C. Scott, Yan Lu, Cynthia Jeffries, Jared T. Hammill, Michele Connelly, Deepak Bhasin, Gloria Holbrook, Lei Yang, Jaeki Min, Courtney Hammill, and Ho Shin Kim
- Subjects
0301 basic medicine ,Lung Neoplasms ,NEDD8 Protein ,Biological Availability ,Antineoplastic Agents ,Pharmacology ,NEDD8 ,Article ,03 medical and health sciences ,Mice ,Structure-Activity Relationship ,0302 clinical medicine ,In vivo ,Cell Line, Tumor ,Proto-Oncogene Proteins ,Drug Discovery ,Structure–activity relationship ,Animals ,Humans ,Urea ,biology ,Drug discovery ,Chemistry ,Intracellular Signaling Peptides and Proteins ,Proteins ,Cullin Proteins ,In vitro ,Squamous carcinoma ,030104 developmental biology ,030220 oncology & carcinogenesis ,Ubiquitin-Conjugating Enzymes ,biology.protein ,Carcinoma, Squamous Cell ,Microsomes, Liver ,Molecular Medicine ,Female ,Neddylation ,Drug Screening Assays, Antitumor ,Cullin - Abstract
We previously reported the discovery, validation, and structure–activity relationships of a series of piperidinyl ureas that potently inhibit the DCN1–UBE2M interaction. We demonstrated that compound 7 inhibits both the DCN1–UBE2M protein–protein interaction and DCN1-mediated cullin neddylation in biochemical assays and reduces levels of steady-state cullin neddylation in a squamous carcinoma cell line harboring DCN1 amplification. Although compound 7 exhibits good solubility and permeability, it is rapidly metabolized in microsomal models (CLint = 170 mL/min/kg). This work lays out the discovery of an orally bioavailable analogue, NAcM-OPT (67). Compound 67 retains the favorable biochemical and cellular activity of compound 7 but is significantly more stable both in vitro and in vivo. Compound 67 is orally bioavailable, well tolerated in mice, and currently used to study the effects of acute pharmacologic inhibition of the DCN1–UBE2M interaction on the NEDD8/CUL pathway.
- Published
- 2018
39. Progress in the Development of Preventative Drugs for Cisplatin-Induced Hearing Loss
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Robert A, Hazlitt, Jaeki, Min, and Jian, Zuo
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Drug Discovery ,Perspective ,Animals ,Humans ,Cisplatin ,Hearing Loss - Abstract
Cisplatin is a highly effective treatment for malignant cancers and has become a cornerstone in chemotherapeutic regimens. Unfortunately, its use in the clinic is often coupled with a high incidence of severe hearing loss. Over the past few decades, enormous effort has been put forth to find protective agents that selectively protect against the ototoxic side effects of cisplatin and do not interfere with its antitumoral activity. Many therapies have been successful in preclinical work, but only a few have shown any protection in the clinic, and none have been approved by the FDA. This review summarizes the clinical and preclinical studies of the most effective small-molecule candidates currently in clinical trials, while also detailing their molecular mechanisms of action, to gain insight for future drug development in the field.
- Published
- 2018
40. CDK2 inhibitors as candidate therapeutics for cisplatin- and noise-induced hearing loss
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Jie Fang, R. Kiplin Guy, Robert A. Hazlitt, Robyn A. Umans, Burgess B. Freeman, Junmin Peng, Jaeki Min, Cheng Cheng, Asli N. Goktug, Duane G. Currier, Richard W. Kriwacki, Shiyong Diao, Luigi I. Iconaru, Marie Morfouace, Justine D. Bonga, Jian Zuo, Martine F. Roussel, Michael R. Taylor, Yinmei Zhou, Tal Teitz, and Taosheng Chen
- Subjects
0301 basic medicine ,Indoles ,Drug Resistance ,Immunology and Allergy ,Zebrafish ,Research Articles ,chemistry.chemical_classification ,Mice, Knockout ,biology ,Cell Death ,Kinase ,3. Good health ,Mitochondria ,Toxicity ,medicine.symptom ,Noise-induced hearing loss ,medicine.drug ,Hearing loss ,Immunology ,Article ,Small Molecule Libraries ,03 medical and health sciences ,Cell Line, Tumor ,Hair Cells, Auditory ,medicine ,otorhinolaryngologic diseases ,Animals ,Kinase activity ,Protein Kinase Inhibitors ,Cisplatin ,Reactive oxygen species ,business.industry ,Cyclin-dependent kinase 2 ,Cyclin-Dependent Kinase 2 ,Benzazepines ,medicine.disease ,Lateral Line System ,Rats ,Mice, Inbred C57BL ,030104 developmental biology ,Germ Cells ,chemistry ,Hearing Loss, Noise-Induced ,Cytoprotection ,biology.protein ,Cancer research ,business ,Reactive Oxygen Species - Abstract
Using a high-throughput small molecule screen, Teitz et al. identify kenpaullone, a cyclin-dependent kinase 2 inhibitor, which when delivered locally confers protection against cisplatin- and noise-induced hearing loss in zebrafish, mice, and rats and reduces mitochondrial ROS production and cochlear cell death., Hearing loss caused by aging, noise, cisplatin toxicity, or other insults affects 360 million people worldwide, but there are no Food and Drug Administration–approved drugs to prevent or treat it. We screened 4,385 small molecules in a cochlear cell line and identified 10 compounds that protected against cisplatin toxicity in mouse cochlear explants. Among them, kenpaullone, an inhibitor of multiple kinases, including cyclin-dependent kinase 2 (CDK2), protected zebrafish lateral-line neuromasts from cisplatin toxicity and, when delivered locally, protected adult mice and rats against cisplatin- and noise-induced hearing loss. CDK2-deficient mice displayed enhanced resistance to cisplatin toxicity in cochlear explants and to cisplatin- and noise-induced hearing loss in vivo. Mechanistically, we showed that kenpaullone directly inhibits CDK2 kinase activity and reduces cisplatin-induced mitochondrial production of reactive oxygen species, thereby enhancing cell survival. Our experiments have revealed the proapoptotic function of CDK2 in postmitotic cochlear cells and have identified promising therapeutics for preventing hearing loss.
- Published
- 2017
41. Discovery of novel, orally bioavailable, antileishmanial compounds using phenotypic screening
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Yizhe Chen, W. Armand Guiguemde, Diana Ortiz, Fangyi Zhu, Kayla Stalheim, Anang A. Shelat, Michele Connelly, Lei Yang, David C. Smithson, Jaeki Min, Alex G. Johnson, Angela K. Carrillo, Carolyn Elya, R. Kiplin Guy, Scott M. Landfear, and Jared T. Hammill
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0301 basic medicine ,Life Cycles ,Chemistry, Pharmaceutical ,Leishmania mexicana ,Drug Evaluation, Preclinical ,Administration, Oral ,Drug resistance ,Protozoology ,Pharmacology ,Mice ,White Blood Cells ,Animal Cells ,Zoonoses ,Drug Discovery ,Medicine and Health Sciences ,Leishmaniasis ,Mice, Inbred BALB C ,Drug discovery ,lcsh:Public aspects of medicine ,Animal Models ,3. Good health ,Phenotype ,Infectious Diseases ,Experimental Organism Systems ,Female ,Protozoan Life Cycles ,Cellular Types ,Research Article ,Amastigotes ,Neglected Tropical Diseases ,medicine.drug ,lcsh:Arctic medicine. Tropical medicine ,lcsh:RC955-962 ,Immune Cells ,Phenotypic screening ,Immunology ,030106 microbiology ,Antiprotozoal Agents ,Leishmaniasis, Cutaneous ,Mouse Models ,Library Screening ,Biology ,Research and Analysis Methods ,Microbiology ,Cell Line ,03 medical and health sciences ,Model Organisms ,Cutaneous leishmaniasis ,In vivo ,Parasitic Diseases ,medicine ,Animals ,Humans ,Pharmacokinetics ,Molecular Biology Techniques ,Amastigote ,Molecular Biology ,Molecular Biology Assays and Analysis Techniques ,Miltefosine ,Blood Cells ,Protozoan Infections ,Macrophages ,Promastigotes ,Public Health, Environmental and Occupational Health ,Biology and Life Sciences ,lcsh:RA1-1270 ,Cell Biology ,Tropical Diseases ,medicine.disease ,biology.organism_classification ,030104 developmental biology ,Developmental Biology - Abstract
Leishmaniasis is a parasitic infection that afflicts approximately 12 million people worldwide. There are several limitations to the approved drug therapies for leishmaniasis, including moderate to severe toxicity, growing drug resistance, and the need for extended dosing. Moreover, miltefosine is currently the only orally available drug therapy for this infection. We addressed the pressing need for new therapies by pursuing a two-step phenotypic screen to discover novel, potent, and orally bioavailable antileishmanials. First, we conducted a high-throughput screen (HTS) of roughly 600,000 small molecules for growth inhibition against the promastigote form of the parasite life cycle using the nucleic acid binding dye SYBR Green I. This screen identified approximately 2,700 compounds that inhibited growth by over 65% at a single point concentration of 10 μM. We next used this 2700 compound focused library to identify compounds that were highly potent against the disease-causing intra-macrophage amastigote form and exhibited limited toxicity toward the host macrophages. This two-step screening strategy uncovered nine unique chemical scaffolds within our collection, including two previously described antileishmanials. We further profiled two of the novel compounds for in vitro absorption, distribution, metabolism, excretion, and in vivo pharmacokinetics. Both compounds proved orally bioavailable, affording plasma exposures above the half-maximal effective concentration (EC50) concentration for at least 12 hours. Both compounds were efficacious when administered orally in a murine model of cutaneous leishmaniasis. One of the two compounds exerted potent activity against trypanosomes, which are kinetoplastid parasites related to Leishmania species. Therefore, this compound could help control multiple parasitic diseases. The promising pharmacokinetic profile and significant in vivo efficacy observed from our HTS hits highlight the utility of our two-step phenotypic screening strategy and strongly suggest that medicinal chemistry optimization of these newly identified scaffolds will lead to promising candidates for an orally available anti-parasitic drug., Author summary Leishmaniasis, caused by the protozoa of the Leishmania species, represents a spectrum of diseases that afflicts roughly 12 million individuals worldwide. Current drug therapies for this parasitic disease are suboptimal because they are toxic, expensive, difficult to administer, and subject to drug resistance. In order to identify new and improved drug candidates, we screened a large library of small molecules for compounds that inhibit parasitic growth inside mammalian host macrophages, and have low toxicity toward the macrophages. We discovered two compounds that significantly impaired disease progression when administered orally in an animal model of cutaneous leishmaniasis. The promising pharmacokinetic and in vivo efficacy profile of the compounds make them attractive starting points for pharmaceutical development.
- Published
- 2017
42. Organocatalytic, Diastereo- and Enantioselective Synthesis of Nonsymmetric cis-Stilbene Diamines: A Platform for the Preparation of Single-Enantiomer cis-Imidazolines for Protein–Protein Inhibition
- Author
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Brandon A. Vara, Jaeki Min, Jeffrey N. Johnston, R. Kiplin Guy, Tyler A. Davis, Vanessa Arredondo, Michael W. Danneman, John C. Tellis, Kristin Finch, and Anand Mayasundari
- Subjects
Aldimine ,Stereochemistry ,Amidines ,Apoptosis ,Fluorescence Polarization ,Stereoisomerism ,Diamines ,010402 general chemistry ,01 natural sciences ,Article ,Catalysis ,Amidine ,chemistry.chemical_compound ,Cell Line, Tumor ,Stilbenes ,Humans ,Protein Interaction Domains and Motifs ,Imidazolines ,Chromatography, High Pressure Liquid ,chemistry.chemical_classification ,Molecular Structure ,010405 organic chemistry ,Aryl ,Organic Chemistry ,Enantioselective synthesis ,Substrate (chemistry) ,Small molecule ,Organic Chemistry Phenomena ,0104 chemical sciences ,chemistry ,Enantiomer - Abstract
The finding by scientists at Hoffmann-La Roche that cis-imidazolines could disrupt the protein–protein interaction between p53 and MDM2, thereby inducing apoptosis in cancer cells, raised considerable interest in this scaffold over the past decade. Initial routes to these small molecules (i.e., Nutlin-3) provided only the racemic form, with enantiomers being enriched by chromatographic separation using high-pressure liquid chromatography (HPLC) and a chiral stationary phase. Reported here is the first application of an enantioselective aza-Henry approach to nonsymmetric cis-stilbene diamines and cis-imidazolines. Two novel mono(amidine) organocatalysts (MAM) were discovered to provide high levels of enantioselection (>95% ee) across a broad range of substrate combinations. Furthermore, the versatility of the aza-Henry strategy for preparing nonsymmetric cis-imidazolines is illustrated by a comparison of the roles of aryl nitromethane and aryl aldimine in the key step, which revealed unique substrate electronic effects providing direction for aza-Henry substrate–catalyst matching. This method was used to prepare highly substituted cis-4,5-diaryl imidazolines that project unique aromatic rings, and these were evaluated for MDM2-p53 inhibition in a fluorescence polarization assay. The diversification of access to cis-stilbene diamine-derived imidazolines provided by this platform should streamline their further development as chemical tools for disrupting protein–protein interactions.
- Published
- 2014
43. Dihydroquinazolinone Inhibitors of Proliferation of Blood and Liver Stage Malaria Parasites
- Author
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W. Armand Guiguemde, Michele Connelly, R. Kiplin Guy, Julie Clark, Jaeki Min, Andreia D. Magalhães, Emily R. Derbyshire, and Jon Clardy
- Subjects
Plasmodium ,Plasmodium berghei ,Plasmodium falciparum ,Drug Evaluation, Preclinical ,Pharmacology ,Antimalarials ,Structure-Activity Relationship ,parasitic diseases ,medicine ,Humans ,Moiety ,Potency ,Structure–activity relationship ,Experimental Therapeutics ,Pharmacology (medical) ,Liver stage ,Life Cycle Stages ,biology ,medicine.disease ,biology.organism_classification ,Malaria ,Infectious Diseases ,Liver ,Toxicity ,Quinazolines - Abstract
Drugs that target both the liver and blood stages of malaria will be needed to reduce the disease's substantial worldwide morbidity and mortality. Evaluation of a 259-member library of compounds that block proliferation of the blood stage of malaria revealed several scaffolds—dihydroquinazolinones, phenyldiazenylpyridines, piperazinyl methyl quinolones, and bis-benzimidazoles—with promising activity against the liver stage. Focused structure-activity studies on the dihydroquinazolinone scaffold revealed several molecules with excellent potency against both blood and liver stages. One promising early lead with dual activity is 2-( p -bromophenyl)-3-(2-(diethylamino)ethyl)-2,3-dihydroquinazolin-4(1 H )-one with 50% effective concentrations (EC 50 s) of 0.46 μM and 0.34 μM against liver stage Plasmodium berghei ANKA and blood stage Plasmodium falciparum 3D7 parasites, respectively. Structure-activity relationships revealed that liver stage activity for this compound class requires a 3-dialkyl amino ethyl group and is abolished by substitution at the ortho -position of the phenyl moiety. These compounds have minimal toxicity to mammalian cells and are thus attractive compounds for further development.
- Published
- 2014
44. Flexibility is important for inhibition of the MDM2/p53 protein-protein interaction by cyclic β-hairpins
- Author
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R. Kiplin Guy, Jaeki Min, Morten Grøtli, M. Brett Waddell, Emma Danelius, Máté Erdélyi, Mariell Pettersson, and Matilda Bred
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Models, Molecular ,Protein Conformation, alpha-Helical ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Protein–protein interaction ,Protein structure ,Physical and Theoretical Chemistry ,Surface plasmon resonance ,Binding site ,Flexibility (engineering) ,biology ,010405 organic chemistry ,Drug discovery ,Chemistry ,Organic Chemistry ,Proto-Oncogene Proteins c-mdm2 ,3. Good health ,0104 chemical sciences ,Drug Design ,biology.protein ,Mdm2 ,Peptidomimetics ,Tumor Suppressor Protein p53 ,Fluorescence anisotropy ,Protein Binding - Abstract
Protein–protein interactions that have large, flat and featureless binding sites are difficult drug targets. In the development of their modulators conventional drug discovery strategies are often unsuccessful. Gaining a detailed understanding of the binding mode of protein–protein interaction inhibitors is therefore of vast importance for their future pharmaceutical use. The MDM2/p53 protein pair is a highly promising target for cancer treatment. Disruption of the protein complex using p53 α-helix mimetics has been shown to be a successful strategy to control p53 activity. To gain further insight into the binding of inhibitors to MDM2, the flexibility of four cyclic β-hairpins that act as α-helical mimetics and potential MDM2/p53 interaction inhibitors was investigated in relation to their inhibitory activity. MDM2-binding of the mimetics was determined using fluorescence polarization and surface plasmon resonance assays, whereas their conformation and dynamics in solution was described by the combined experimental and computational NAMFIS analysis. Molecular flexibility was shown to be important for the activity of the cyclic β-hairpin based MDM2 inhibitors.
- Published
- 2016
45. Hit-to-Lead Studies for the Antimalarial Tetrahydroisoquinolone Carboxanilides
- Author
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Steve Castro, Julie Clark, Amy Matheny, David M. Floyd, Philip D. Stein, Spencer Knapp, Gloria Holbrook, Martina Sigal, Fangyi Zhu, Senthil Krishnan, Michele Connelly, Jian Liu, Zheng Wang, Sridevi Bashyum, Rajkumar Dhinakaran, Jaeki Min, and R. Kiplin Guy
- Subjects
Erythrocytes ,Plasmodium falciparum ,Drug resistance ,Pharmacology ,010402 general chemistry ,01 natural sciences ,Article ,Antimalarials ,Mice ,Structure-Activity Relationship ,In vivo ,Drug Discovery ,Potency ,Animals ,Humans ,Anilides ,Cytotoxicity ,biology ,010405 organic chemistry ,Chemistry ,Stereoisomerism ,Hit to lead ,biology.organism_classification ,Isoquinolines ,In vitro ,Coculture Techniques ,0104 chemical sciences ,Solubility ,Microsomes, Liver ,Molecular Medicine ,Enantiomer - Abstract
Phenotypic whole-cell screening in erythrocytic cocultures of Plasmodium falciparum identified a series of dihydroisoquinolones that possessed potent antimalarial activity against multiple resistant strains of P. falciparum in vitro and show no cytotoxicity to mammalian cells. Systematic structure–activity studies revealed relationships between potency and modifications at N-2, C-3, and C-4. Careful structure–property relationship studies, coupled with studies of metabolism, addressed the poor aqueous solubility and metabolic vulnerability, as well as potential toxicological effects, inherent in the more potent primary screening hits such as 10b. Analogues 13h and 13i, with structural modifications at each site, were shown to possess excellent antimalarial activity in vivo. The (+)-(3S,4S) enantiomer of 13i and similar analogues were identified as the more potent. On the basis of these studies, we have selected (+)-13i for further study as a preclinical candidate.
- Published
- 2016
46. Monitoring ligand-induced protein ordering in drug discovery
- Author
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Jaeki Min, Kristin Finch, Richard W. Kriwacki, Michael A. Dyer, David Ban, Lie Min, R. Kiplin Guy, Lyra Griffiths, Christy R. Grace, Nagakumar Bharatham, Anand Mayasundari, and Donald Bashford
- Subjects
0301 basic medicine ,Models, Molecular ,MDMX ,Magnetic Resonance Spectroscopy ,Stereochemistry ,Protein Conformation ,Plasma protein binding ,010402 general chemistry ,01 natural sciences ,Article ,Piperazines ,03 medical and health sciences ,Structure-Activity Relationship ,Protein structure ,Structural Biology ,Cell Line, Tumor ,Drug Discovery ,Structure–activity relationship ,Humans ,Molecular Biology ,Drug discovery ,Chemistry ,Imidazoles ,Proto-Oncogene Proteins c-mdm2 ,Ligand (biochemistry) ,Small molecule ,0104 chemical sciences ,030104 developmental biology ,Protein Multimerization ,Tumor Suppressor Protein p53 ,Heteronuclear single quantum coherence spectroscopy ,Protein Binding - Abstract
While the gene for p53 is mutated in many human cancers causing loss of function, many others maintain a wild-type gene but exhibit reduced p53 tumor suppressor activity through overexpression of the negative regulators, Mdm2 and/or MdmX. For the latter mechanism of loss of function, the activity of endogenous p53 can be restored through inhibition of Mdm2 or MdmX with small molecules. We previously reported a series of compounds based upon the Nutlin-3 chemical scaffold that bind to both MdmX and Mdm2 [Vara, B. A. et al. (2014) Organocatalytic, diastereo- and enantioselective synthesis of nonsymmetric cis-stilbene diamines: A platform for the preparation of single-enantiomer cis-imidazolines for protein-protein inhibition. J. Org. Chem. 79, 6913-6938]. Here we present the first solution structures based on data from NMR spectroscopy for MdmX in complex with four of these compounds and compare them with the MdmX:p53 complex. A p53-derived peptide binds with high affinity (Kd value of 150nM) and causes the formation of an extensive network of hydrogen bonds within MdmX; this constitutes the induction of order within MdmX through ligand binding. In contrast, the compounds bind more weakly (Kd values from 600nM to 12μM) and induce an incomplete hydrogen bond network within MdmX. Despite relatively weak binding, the four compounds activated p53 and induced p21(Cip1) expression in retinoblastoma cell lines that overexpress MdmX, suggesting that they specifically target MdmX and/or Mdm2. Our results document structure-activity relationships for lead-like small molecules targeting MdmX and suggest a strategy for their further optimization in the future by using NMR spectroscopy to monitor small-molecule-induced protein order as manifested through hydrogen bond formation.
- Published
- 2016
47. Design, Synthesis and Evaluation of 2,5-Diketopiperazines as Inhibitors of the MDM2-p53 Interaction
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Jaeki Min, M. Brett Waddell, Maria Quant, R. Kiplin Guy, Richard W. Kriwacki, Mariell Pettersson, Kristina Luthman, Luigi I. Iconaru, and Morten Grøtli
- Subjects
Models, Molecular ,Stereochemistry ,lcsh:Medicine ,Peptide ,Chemistry Techniques, Synthetic ,Diketopiperazines ,Protein Structure, Secondary ,Protein–protein interaction ,chemistry.chemical_compound ,Protein structure ,Peptide synthesis ,Humans ,Binding site ,lcsh:Science ,chemistry.chemical_classification ,Multidisciplinary ,Binding Sites ,lcsh:R ,Rational design ,Proto-Oncogene Proteins c-mdm2 ,Biochemistry ,chemistry ,Docking (molecular) ,Drug Design ,lcsh:Q ,Tumor Suppressor Protein p53 ,Research Article ,Protein Binding - Abstract
The transcription factor p53 is the main tumour suppressor in cells and many cancer types have p53 mutations resulting in a loss of its function. In tumours that retain wild-type p53 function, p53 activity is down-regulated by MDM2 (human murine double minute 2) via a direct protein-protein interaction. We have designed and synthesised two series of 2,5-diketopiperazines as inhibitors of the MDM2-p53 interaction. The first set was designed to directly mimic the alpha-helical region of the p53 peptide, containing key residues in the i, i+4 and i+7 positions of a natural alpha-helix. Conformational analysis indicated that 1,3,6-trisubstituted 2,5-diketopiperazines were able to place substituents in the same spatial orientation as an alpha-helix template. The key step of the synthesis involved the cyclisation of substituted dipeptides. The other set of tetrasubstituted 2,5-diketopiperazines were designed based on structure-based docking studies and the Ugi multicomponent reaction was used for the synthesis. This latter set comprised the most potent inhibitors which displayed micromolar IC50 values in a biochemical fluorescence polarisation assay.
- Published
- 2015
48. Development of a High-Affinity Inhibitor of the Prostaglandin Transporter
- Author
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Jaeki Min, Michael P. Lisanti, Jean-Francois Jasmin, Young-Tae Chang, Victor L. Schuster, and Yuling Chi
- Subjects
Male ,medicine.medical_specialty ,Organic anion transporter 1 ,medicine.medical_treatment ,Organic Anion Transporters ,Endogeny ,Dinoprostone ,Cell Line ,Rats, Sprague-Dawley ,Inhibitory Concentration 50 ,Mice ,Structure-Activity Relationship ,Drug Discovery and Translational Medicine ,chemistry.chemical_compound ,Dogs ,Internal medicine ,para-Aminobenzoates ,medicine ,Extracellular ,Animals ,Molecular Targeted Therapy ,Benzamide ,Ductus Arteriosus, Patent ,Prostaglandin-E Synthases ,Mice, Knockout ,Pharmacology ,biology ,PROSTAGLANDIN TRANSPORTER ,Triazines ,Biological Transport ,Metabolism ,Control Groups ,High-Throughput Screening Assays ,Rats ,Intramolecular Oxidoreductases ,Endocrinology ,chemistry ,Prostaglandin-Endoperoxide Synthases ,biology.protein ,Molecular Medicine ,lipids (amino acids, peptides, and proteins) ,Signal transduction ,4-Aminobenzoic Acid ,Signal Transduction ,Prostaglandin E - Abstract
Prostaglandin E(2) (PGE(2)) triggers a vast array of biological signals and physiological events. The prostaglandin transporter (PGT) controls PGE(2) influx and is rate-limiting for PGE(2) metabolism and signaling termination. PGT global knockout mice die on postnatal day 1 from patent ductus arteriosus. A high-affinity PGT inhibitor would thus be a powerful tool for studying PGT function in adult animals. Moreover, such an inhibitor could be potentially developed into a therapeutic drug targeting PGT. Based on structure-activity relationship studies that built on recently identified inhibitors of PGT, we obtained N-(2-(2-(2-azidoethoxy)ethoxy)ethyl)-4-((4-((2-(2-(2-benzamidoethoxy)ethoxy)ethyl)amino)-6-((4-hydroxyphenyl)amino)-1,3,5-triazin-2-yl)amino)benzamide (T26A), a competitive inhibitor of PGT, with a K(i) of 378 nM. T26A seems to be highly selective for PGT, because it neither interacts with a PGT homolog in the organic anion transporter family nor affects PGE(2) synthesis. In Madin-Darby canine kidney cells stably transfected with PGT, T26A blocked PGE(2) metabolism, resulting in retention of PGE(2) in the extracellular compartment and the negligible appearance of PGE(2) metabolites in the intracellular compartment. Compared with vehicle, T26A injected intravenously into rats effectively doubled the amount of endogenous PGE(2) in the circulation and reduced the level of circulating endogenous PGE(2) metabolites to 50%. Intravenous T26A was also able to slow the metabolism of exogenously injected PGE(2). These results confirm that PGT directly regulates PGE(2) metabolism and demonstrate that a high-affinity inhibitor of PGT can effectively prevent PGE(2) metabolism and prolong the half-life of circulating PGE(2).
- Published
- 2011
49. Blocking eIF4E-eIF4G Interaction as a Strategy To Impair Coronavirus Replication
- Author
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Jaeki Min, Haian Fu, Dima Kozakov, Yuhong Du, David R. Hall, Pierre J. Talbot, Regina Cencic, Raymond Dingledine, Sandor Vajda, Marc Desforges, Jerry Pelletier, Department of Biochemistry [Montréal], McGill University = Université McGill [Montréal, Canada], Institut Armand Frappier (INRS-IAF), Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS), Department of Biomedical Engineering [Boston], Boston University [Boston] (BU), Emory Chemical Biology Discovery Center, Emory University [Atlanta, GA], Goodman Cancer Research Center [Montréal] (GCRC), and Fellowship support was from a CIHR Cancer Consortium Training Grant Award and a Cole Foundation Award to R.C. This work was supported by grants from the National Institutes of Health (1 R01 CA114475 and MH081216-01) and the Canadian Cancer Society Research Institute (17099) to J.P., a grant MT-9203 from the Institute of Infection and Immunity, Canadian Institutes of Health Research, to P.J.T., and NIH PHS 5U54 HG003918 to R.D. and H.F.
- Subjects
MESH: Antiviral Agents ,RNA Caps ,MESH: High-Throughput Screening Assays ,Human coronavirus 229E ,viruses ,Immunology ,Virus Replication ,medicine.disease_cause ,Antiviral Agents ,Microbiology ,Cell Line ,Small Molecule Libraries ,MESH: RNA Caps ,Viral Proteins ,Eukaryotic initiation factor 4F ,chemistry.chemical_compound ,MESH: Drug Discovery ,MESH: Small Molecule Libraries ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,Coronavirus 229E, Human ,Virology ,Drug Discovery ,Vaccines and Antiviral Agents ,medicine ,Humans ,Coronavirus ,Subgenomic mRNA ,MESH: Humans ,biology ,EIF4G ,MESH: Virus Replication ,EIF4E ,MESH: Eukaryotic Initiation Factor-4G ,EIF4A1 ,MESH: Coronavirus 229E, Human ,MESH: Eukaryotic Initiation Factor-4E ,biology.organism_classification ,MESH: Viral Proteins ,MESH: Cell Line ,High-Throughput Screening Assays ,Eukaryotic Initiation Factor-4E ,chemistry ,Viral replication ,MESH: Protein Biosynthesis ,Protein Biosynthesis ,Insect Science ,Eukaryotic Initiation Factor-4G - Abstract
Coronaviruses are a family of enveloped single-stranded positive-sense RNA viruses causing respiratory, enteric, and neurologic diseases in mammals and fowl. Human coronaviruses are recognized to cause up to a third of common colds and are suspected to be involved in enteric and neurologic diseases. Coronavirus replication involves the generation of nested subgenomic mRNAs (sgmRNAs) with a common capped 5′ leader sequence. The translation of most of the sgmRNAs is thought to be cap dependent and displays a requirement for eukaryotic initiation factor 4F (eIF4F), a heterotrimeric complex needed for the recruitment of 40S ribosomes. We recently reported on an ultrahigh-throughput screen to discover compounds that inhibit eIF4F activity by blocking the interaction of two of its subunits (R. Cencic et al., Proc. Natl. Acad. Sci. U. S. A. 108: 1046–1051, 2011). Herein we describe a molecule from this screen that prevents the interaction between eIF4E (the cap-binding protein) and eIF4G (a large scaffolding protein), inhibiting cap-dependent translation. This inhibitor significantly decreased human coronavirus 229E (HCoV-229E) replication, reducing the percentage of infected cells and intra- and extracellular infectious virus titers. Our results support the strategy of targeting the eIF4F complex to block coronavirus infection.
- Published
- 2011
50. Antischistosomal versus Antiandrogenic Properties of Aryl Hydantoin Ro 13-3978
- Author
-
Qingjie Zhao, R. Kiplin Guy, Chunkai Wang, Mireille Vargas, Ming-Fong Lin, Yuxiang Dong, Xiaofang Wang, Jennifer Keiser, Jonathan L. Vennerstrom, Sakthivel Muniyan, and Jaeki Min
- Subjects
Male ,medicine.medical_specialty ,Hydantoin ,Biology ,Imidazolidines ,Mice ,Schistosomicides ,chemistry.chemical_compound ,Cell Line, Tumor ,Virology ,Internal medicine ,Androgen Receptor Antagonists ,medicine ,Animals ,Humans ,Cell Proliferation ,Cell growth ,Hydantoins ,Dihydrotestosterone ,Articles ,Schistosoma mansoni ,biology.organism_classification ,Ligand (biochemistry) ,Schistosomiasis mansoni ,Androgen receptor ,Disease Models, Animal ,Infectious Diseases ,Endocrinology ,chemistry ,Nilutamide ,Parasitology ,medicine.drug - Abstract
In the early 1980s, the antischistosomal aryl hydantoin Ro 13-3978 (AH01), a close structural analogue of the androgen receptor antagonist nilutamide, was discovered. Administration of 100 mg/kg oral doses of AH01 to mice infected with adult and juvenile Schistosoma mansoni produced 95% and 64% total worm burden reductions, confirming its high activity against adult worms, and showing that AH01 is also effective against juvenile infections. AH01 had no measureable interaction with the androgen receptor in a ligand competition assay, but it did block dihydrotestosterone-induced cell proliferation in an androgen-dependent human prostate cancer cell line. For AH01, nilutamide, and three closely related aryl hydantoin derivatives, there was no correlation between antischistosomal activity and androgen receptor interaction.
- Published
- 2014
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