23 results on '"Natalia Matassova"'
Search Results
2. PAC-FragmentDEL – photoactivated covalent capture of DNA-encoded fragments for hit discovery
- Author
-
Huiyong Ma, James B. Murray, Huadong Luo, Xuemin Cheng, Qiuxia Chen, Chao Song, Cong Duan, Ping Tan, Lifang Zhang, Jian Liu, Barry A. Morgan, Jin Li, Jinqiao Wan, Lisa M. Baker, William Finnie, Lucie Guetzoyan, Richard Harris, Nicole Hendrickson, Natalia Matassova, Heather Simmonite, Julia Smith, Roderick E. Hubbard, and Guansai Liu
- Subjects
Pharmacology ,Organic Chemistry ,Drug Discovery ,Pharmaceutical Science ,Molecular Medicine ,Biochemistry - Abstract
We describe a novel approach for screening fragments against a protein that combines the sensitivity of DNA-encoded library technology with the ability of fragments to explore what will bind. Each of the members of the library consists of a fragment which is linked to a photoactivatable diazirine moiety. Split and pool synthesis combines each fragment with a set of linkers with the version of the library reported here containing some 70k different compounds, each with an individual DNA code. Incubation of the library with a protein sample is followed by photoactivation, washing and subsequent PCR and sequencing which allows the individual fragment hits to be identified. We illustrate how the approach allows successful hit fragment identification using only microgram quantities of material for two targets. PAK4 is a kinase for which conventional fragment screening has generated many advance leads. The as yet undrugged target, 2-epimerase, presents a more challenging active site for identification of hit compounds. In both cases, PAC-FragmentDEL identified fragments validated as hits by ligand-observed NMR measurements and crystal structure determination of off-DNA sample binding to the proteins.
- Published
- 2022
3. Establishing Drug Discovery and Identification of Hit Series for the Anti-apoptotic Proteins, Bcl-2 and Mcl-1
- Author
-
Pawel Dokurno, R. Harris, Patrick Casara, James Edward Paul Davidson, Olivier Geneste, Julia Smith, Douglas S. Williamson, Natalia Matassova, Yikang Wang, Allan M. Jordan, Stephen D. Roughley, András Kotschy, Roderick E. Hubbard, Jerome Stark, John A. Hickman, Chen I-Jen, Ben Davis, James Brooke Murray, C. Pedder, Walmsley Claire, Thierry Le Diguarher, Neil Whitehead, Stuart C. Ray, and Christopher John Graham
- Subjects
Series (mathematics) ,Chemistry ,Drug discovery ,General Chemical Engineering ,Isothermal titration calorimetry ,General Chemistry ,Computational biology ,Small molecule ,Article ,Anti-Apoptotic Proteins ,lcsh:Chemistry ,Heteronuclear molecule ,lcsh:QD1-999 ,Surface plasmon resonance - Abstract
We describe our work to establish structure- and fragment-based drug discovery to identify small molecules that inhibit the anti-apoptotic activity of the proteins Mcl-1 and Bcl-2. This identified hit series of compounds, some of which were subsequently optimized to clinical candidates in trials for treating various cancers. Many protein constructs were designed to identify protein with suitable properties for different biophysical assays and structural methods. Fragment screening using ligand-observed NMR experiments identified several series of compounds for each protein. The series were assessed for their potential for subsequent optimization using 1H and 15N heteronuclear single-quantum correlation NMR, surface plasmon resonance, and isothermal titration calorimetry measurements to characterize and validate binding. Crystal structures could not be determined for the early hits, so NMR methods were developed to provide models of compound binding to guide compound optimization. For Mcl-1, a benzodioxane/benzoxazine series was optimized to a Kd of 40 μM before a thienopyrimidine hit series was identified which subsequently led to the lead series from which the clinical candidate S 64315 (MIK 665) was identified. For Bcl-2, the fragment-derived series were difficult to progress, and a compound derived from a published tetrahydroquinone compound was taken forward as the hit from which the clinical candidate (S 55746) was obtained. For both the proteins, the work to establish a portfolio of assays gave confidence for identification of compounds suitable for optimization.
- Published
- 2019
4. The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models
- Author
-
James A. H. Murray, Fabien Melchiore, Natalia Matassova, C. Pedder, Frédéric Colland, Catherine Chang, Levente Ondi, Laura C. A. Galbraith, Pawel Dokurno, Andrew W. Roberts, Chris D. Riffkin, Nolwen Guigal-Stephan, Proszenyák Ágnes, John A. Hickman, Andreas Strasser, Aurélie Studeny, Gaëlle Lysiak-Auvity, András Kotschy, Gemma L. Kelly, Fabienne Gravé, Ghislaine Guasconi, Brandon J. Aubrey, Attila Paczal, Julia Smith, Szlávik Zoltán, Gabor Radics, Brian Lockhart, David C.S. Huang, Maïa Chanrion, Gaëtane Le Toumelin-Braizat, Jia-Nan Gong, Nicolas Cauquil, Szabolcs Sipos, Alan P. Robertson, Giovanna Pomilio, James Edward Paul Davidson, Zoltán B. Szabó, Ana Leticia Maragno, Chris Graham, Marco J Herold, Gábor Blasko, Margs S. Brennan, Anne-Marie Girard, Allan E. Surgenor, Andrew H. Wei, Csékei Márton, Olivier Geneste, Chen I-Jen, Alain Bruno, David J. Segal, Guillaume Lessene, Balázs Bálint, and Donia M Moujalled
- Subjects
Male ,Models, Molecular ,0301 basic medicine ,Lymphoma ,Antineoplastic Agents ,Apoptosis ,Bcl-xL ,Thiophenes ,Pharmacology ,Models, Biological ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Bcl-2-associated X protein ,Cell Line, Tumor ,Neoplasms ,medicine ,Animals ,Humans ,MCL1 ,bcl-2-Associated X Protein ,Leukemia ,Multidisciplinary ,biology ,Cancer ,medicine.disease ,Xenograft Model Antitumor Assays ,Myeloid Cell Leukemia Sequence 1 Protein ,Pyrimidines ,bcl-2 Homologous Antagonist-Killer Protein ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer cell ,biology.protein ,Female ,Multiple Myeloma ,Bcl-2 Homologous Antagonist-Killer Protein - Abstract
Avoidance of apoptosis is critical for the development and sustained growth of tumours. The pro-survival protein myeloid cell leukemia 1 (MCL1) is overexpressed in many cancers, but the development of small molecules targeting this protein that are amenable for clinical testing has been challenging. Here we describe S63845, a small molecule that specifically binds with high affinity to the BH3-binding groove of MCL1. Our mechanistic studies demonstrate that S63845 potently kills MCL1-dependent cancer cells, including multiple myeloma, leukaemia and lymphoma cells, by activating the BAX/BAK-dependent mitochondrial apoptotic pathway. In vivo, S63845 shows potent anti-tumour activity with an acceptable safety margin as a single agent in several cancers. Moreover, MCL1 inhibition, either alone or in combination with other anti-cancer drugs, proved effective against several solid cancer-derived cell lines. These results point towards MCL1 as a target for the treatment of a wide range of tumours.
- Published
- 2016
5. S55746 is a novel orally active BCL-2 selective and potent inhibitor that impairs hematological tumor growth
- Author
-
Patrick Casara, James Davidson, Audrey Claperon, Gaëtane Le Toumelin-Braizat, Meike Vogler, Alain Bruno, Maïa Chanrion, Gaëlle Lysiak-Auvity, Thierry Le Diguarher, Jérôme-Benoît Starck, Ijen Chen, Neil Whitehead, Christopher Graham, Natalia Matassova, Pawel Dokurno, Christopher Pedder, Youzhen Wang, Shumei Qiu, Anne-Marie Girard, Emilie Schneider, Fabienne Gravé, Aurélie Studeny, Ghislaine Guasconi, Francesca Rocchetti, Sophie Maïga, Jean-Michel Henlin, Frédéric Colland, Laurence Kraus-Berthier, Steven Le Gouill, Martin J.S. Dyer, Roderick Hubbard, Mike Wood, Martine Amiot, Gerald M Cohen, John A. Hickman, Erick Morris, James Murray, Olivier Geneste, Institut de Recherches Servier, Vernalis (R&D) Ltd, Institute for Experimental Cancer Research in Pediatrics, Goethe-Universität Frankfurt am Main, Institut de Recherches Internationales Servier [Suresnes] (IRIS), Novartis Institute of Biomedical Research, Oncology Drug Discovery, Regulation of Bcl2 and p53 Networks in Multiple Myeloma and Mantle Cell Lymphoma (CRCINA-ÉQUIPE 10), Centre de Recherche en Cancérologie et Immunologie Nantes-Angers (CRCINA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Centre National de la Recherche Scientifique (CNRS)-Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Centre National de la Recherche Scientifique (CNRS)-Université d'Angers (UA), Ernest and Helen Scott Haematological Research Institute, University of Leicester, Institute of Translational Medicine, University of Liverpool, Goethe-University Frankfurt, Institut de Recherches Internationales Servier [Suresnes] ( IRIS ), Regulation of Bcl2 and p53 networks in Multiple Myeloma and Mantle Cell Lymphoma ( CRCINA - Département NOHMAD - Equipe 10 ), Centre de recherche de Cancérologie et d'Immunologie / Nantes - Angers ( CRCINA ), Université d'Angers ( UA ) -Université de Nantes ( UN ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche en Santé de l'Université de Nantes ( IRS-UN ) -Centre hospitalier universitaire de Nantes ( CHU Nantes ) -Université d'Angers ( UA ) -Université de Nantes ( UN ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche en Santé de l'Université de Nantes ( IRS-UN ) -Centre hospitalier universitaire de Nantes ( CHU Nantes ), Université d'Angers (UA)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université d'Angers (UA)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre hospitalier universitaire de Nantes (CHU Nantes), and Bernardo, Elizabeth
- Subjects
0301 basic medicine ,apoptosis ,BCL-2 ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,BH3-mimetics ,[ SDV.CAN ] Life Sciences [q-bio]/Cancer ,3. Good health ,inhibitor ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,[SDV.CAN] Life Sciences [q-bio]/Cancer ,Oncology ,030220 oncology & carcinogenesis ,hematological malignancies ,Research Paper - Abstract
International audience; Escape from apoptosis is one of the major hallmarks of cancer cells. The B-cell Lymphoma 2 (BCL-2) gene family encodes pro-apoptotic and anti-apoptotic proteins that are key regulators of the apoptotic process. Overexpression of the pro-survival member BCL-2 is a well-established mechanism contributing to oncogenesis and chemoresistance in several cancers, including lymphoma and leukemia. Thus, BCL-2 has become an attractive target for therapeutic strategy in cancer, as demonstrated by the recent approval of ABT-199 (Venclexta™) in relapsed or refractory Chronic Lymphocytic Leukemia with 17p deletion. Here, we describe a novel orally bioavailable BCL-2 selective and potent inhibitor called S55746 (also known as BCL201). S55746 occupies the hydrophobic groove of BCL-2. Its selectivity profile demonstrates no significant binding to MCL-1, BFL-1 (BCL2A1/A1) and poor affinity for BCL-XL. Accordingly, S55746 has no cytotoxic activity on BCL-XL-dependent cells, such as platelets. In a panel of hematological cell lines, S55746 induces hallmarks of apoptosis including externalization of phosphatidylserine, caspase-3 activation and PARP cleavage. Ex vivo, S55746 induces apoptosis in the low nanomolar range in primary Chronic Lymphocytic Leukemia and Mantle Cell Lymphoma patient samples. Finally, S55746 administered by oral route daily in mice demonstrated robust anti-tumor efficacy in two hematological xenograft models with no weight lost and no change in behavior. Taken together, these data demonstrate that S55746 is a novel, welltolerated BH3-mimetic targeting selectively and potently the BCL-2 protein.
- Published
- 2018
6. VER-246608, a novel pan-isoform ATP competitive inhibitor of pyruvate dehydrogenase kinase, disrupts Warburg metabolism and induces context-dependent cytostasis in cancer cells
- Author
-
Anna Staniszewska, Lisa Baker, Jonathan D. Moore, James Murray, Patrick C. Mahon, Terence Shaw, Paul Brough, Alan Surgenor, Michael Wood, Ben Davis, Jalanie D’Alessandro, Natalia Matassova, and Macias Alba
- Subjects
Models, Molecular ,Pyruvate decarboxylation ,Pyruvate dehydrogenase lipoamide kinase isozyme 1 ,Pyruvate dehydrogenase kinase ,Nov3r ,Protein Serine-Threonine Kinases ,Pyruvate dehydrogenase phosphatase ,Biology ,Binding, Competitive ,chemistry.chemical_compound ,Adenosine Triphosphate ,Tumor Cells, Cultured ,Humans ,Neoplasm Invasiveness ,Glycolysis ,Enzyme Inhibitors ,Molecular Structure ,Pyruvate Dehydrogenase Acetyl-Transferring Kinase ,glycolysis ,Pyruvate dehydrogenase complex ,Cytostasis ,Isoenzymes ,Pyrimidines ,Oncology ,chemistry ,Biochemistry ,Benzamides ,Warburg metabolism ,K562 Cells ,Adenosine triphosphate ,Research Paper - Abstract
Pyruvate dehydrogenase kinase (PDK) is a pivotal enzyme in cellular energy metabolism that has previously been implicated in cancer through both RNAi based studies and clinical correlations with poor prognosis in several cancer types. Here, we report the discovery of a novel and selective ATP competitive pan-isoform inhibitor of PDK, VER-246608. Consistent with a PDK mediated MOA, VER-246608 increased pyruvate dehydrogenase complex (PDC) activity, oxygen consumption and attenuated glycolytic activity. However, these effects were only observed under D-glucose-depleted conditions and required almost complete ablation of PDC E1α subunit phosphorylation. VER-246608 was weakly anti-proliferative to cancer cells in standard culture media; however, depletion of either serum or combined D-glucose/L-glutamine resulted in enhanced cellular potency. Furthermore, this condition-selective cytostatic effect correlated with reduced intracellular pyruvate levels and an attenuated compensatory response involving deamination of L-alanine. In addition, VER-246608 was found to potentiate the activity of doxorubicin. In contrast, the lipoamide site inhibitor, Nov3r, demonstrated sub-maximal inhibition of PDK activity and no evidence of cellular activity. These studies suggest that PDK inhibition may be effective under the nutrient-depleted conditions found in the tumour microenvironment and that combination treatments should be explored to reveal the full potential of this therapeutic strategy.
- Published
- 2014
7. Application of Off-Rate Screening in the Identification of Novel Pan-Isoform Inhibitors of Pyruvate Dehydrogenase Kinase
- Author
-
Stephen Stokes, Patrick C. Mahon, Alan D. Robertson, Jalanie D’Alessandro, Paul Webb, Charles Parry, Lisa Baker, Natalia Matassova, Nicholas G. M. Davies, Sean McKenna, Andrew Massey, Rachel Parsons, Yikang Wang, Macias Alba, Paul Brough, Ben Davis, Michael Wood, Simon Bedford, Jonathan D. Moore, Christopher J. Northfield, Loic le Strat, Stephen D. Roughley, Daniel Maddox, Seema Chavda, James Brooke Murray, Allan E. Surgenor, Johannes W. G. Meissner, Terry Shaw, Heather Simmonite, Stefaniak Emma Jayne, Victoria Chell, Neil Whitehead, and Kirsten Brown
- Subjects
0301 basic medicine ,Male ,Models, Molecular ,Pyruvate dehydrogenase lipoamide kinase isozyme 1 ,Pyruvate dehydrogenase kinase ,Pyruvate dehydrogenase phosphatase ,Protein Serine-Threonine Kinases ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Adenosine Triphosphate ,Cell Line, Tumor ,Drug Discovery ,Transferase ,Humans ,Protein Isoforms ,HSP90 Heat-Shock Proteins ,Phosphorylation ,Protein Kinase Inhibitors ,chemistry.chemical_classification ,Prostatic Neoplasms ,Pyruvate Dehydrogenase Acetyl-Transferring Kinase ,Hit to lead ,Molecular biology ,030104 developmental biology ,Enzyme ,chemistry ,Biochemistry ,030220 oncology & carcinogenesis ,Drug Design ,Molecular Medicine ,Adenosine triphosphate - Abstract
Libraries of nonpurified resorcinol amide derivatives were screened by surface plasmon resonance (SPR) to determine the binding dissociation constant (off-rate, kd) for compounds binding to the pyruvate dehydrogenase kinase (PDHK) enzyme. Parallel off-rate measurements against HSP90 and application of structure-based drug design enabled rapid hit to lead progression in a program to identify pan-isoform ATP-competitive inhibitors of PDHK. Lead optimization identified selective sub-100-nM inhibitors of the enzyme which significantly reduced phosphorylation of the E1α subunit in the PC3 cancer cell line in vitro.
- Published
- 2017
8. Abstract 4482: S64315 (MIK665) is a potent and selective Mcl1 inhibitor with strong antitumor activity across a diverse range of hematologic tumor models
- Author
-
Audrey Clapéron, Balázs Bálint, Chen I-Jen, Alain Bruno, Attila Paczal, Szlávik Zoltán, Zoltán B. Szabó, Ensar Halilovic, Heiko Maacke, Alix Derreal, James Edward Paul Davidson, Maïa Chanrion, Ana Leticia Maragno, Szabolcs Sipos, Fabienne Grave, Olivier Geneste, James Murray, Proszenyák Ágnes, Youzhen Wang, Natalia Matassova, Pawel Dokurno, Allan E. Surgenor, Csékei Márton, Prakash Mistry, András Kotschy, Gaëtane Le Toumelin-Braizat, Erick Morris, Frédéric Colland, Anne-Marie Girard, and Gaëlle Lysiak-Auvity
- Subjects
0301 basic medicine ,Cancer Research ,business.industry ,Venetoclax ,Cancer ,Myeloid leukemia ,medicine.disease ,Lymphoma ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,Cell killing ,Oncology ,chemistry ,Apoptosis ,In vivo ,030220 oncology & carcinogenesis ,Cancer cell ,medicine ,Cancer research ,business - Abstract
Mcl-1 is highly expressed in a variety of human cancers (including those of hematopoietic and lymphoid origin) and is exploited by cancer cells to evade cell death and to develop resistance to diverse chemotherapeutic agents. We disclose, for the first time, the structure of S64315 (also named MIK665) a highly potent and selective inhibitor of Mcl-1 with improved potency over its predecessor S63845 (Kotschy et al, Nature, 2016). S64315/MIK665 is currently in phase 1 in AML (Acute Myeloid Leukemia) and MDS (Myelodysplastic Syndrome) (EudraCT 2016-003768-38, NCT 02979366) and in MM (Multiple Myeloma) and lymphoma (NCT02992483). A fragment-based, structure-guided drug discovery effort led to the identification of S64315/MIK665 that binds to human Mcl-1 with a sub-nanomolar affinity (Ki 0.048 nM) and selectively over other anti-apoptotic Bcl-2 family members. It has similar affinity for human, rat, dog and monkey Mcl-1 but about a ten-fold lower affinity for mouse Mcl-1. S64315/MIK665 causes dose-dependent activation of the intrinsic apoptosis pathway in a Bax/Bak-dependent manner, as measured by increased caspase activity and cleaved PARP. S64315/MIK665 shows strong cell killing activity in a diverse panel of human hematological tumor cell lines, including AML, lymphoma and MM. The activity profile of S64315/MIK665 is distinct from that of venetoclax, a selective Bcl2 inhibitor. In vivo, S64315 as single agent demonstrated potent and dose-dependent apoptotic and antitumor response after intravenous administration in several human hematological tumor models grafted in immuno-compromised mice and rats. Complete regression of established tumors, at well tolerated doses, was achieved using different intravenous dosing regimens in rats as well as in mice. Finally, dual BH3-mimetic targeting approach combining S64315/MIK665 with BCL2 inhibitors showed strong and durable antitumor responses in several hematological tumor models both in vitro and in vivo. Citation Format: Ana Leticia Maragno, Prakash Mistry, András Kotschy, Zoltán Szlavik, James Murray, James Davidson, Gaëtane Le Toumelin-Braizat, Maïa Chanrion, Alain Bruno, Audrey Claperon, Heiko Maacke, Erick Morris, Youzhen Wang, Alix Derreal, Márton Csekei, Attila Paczal, Zoltán Szabo, Szabolcs Sipos, Agnes Proszenyak, Balázs Balint, Allan Surgenor, Pawel Dokurno, Natalia Matassova, Ijen Chen, Gaëlle Lysiak-Auvity, Anne-Marie Girard, Fabienne Grave, Frédéric Colland, Ensar Halilovic, Olivier Geneste. S64315 (MIK665) is a potent and selective Mcl1 inhibitor with strong antitumor activity across a diverse range of hematologic tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4482.
- Published
- 2019
9. Targeting conserved water molecules: Design of 4-aryl-5-cyanopyrrolo[2,3-d]pyrimidine Hsp90 inhibitors using fragment-based screening and structure-based optimization
- Author
-
Andrew Massey, Stephanie Geoffrey, Stuart C. Ray, Hart Terance William, Joseph Schoepfer, Michael Wood, Nicholas G. M. Davies, James A. H. Murray, Paul Webb, Michael Rugaard Jensen, Roderick E. Hubbard, Nicolas Foloppe, Jonathan D. Moore, Lisa Wright, Allan E. Surgenor, Robert M. Pratt, Kirsten Scriven, Alan D. Robertson, Stephen Stokes, Paul Brough, Howard Langh Am Mansell, Martin J. Drysdale, Natalia Matassova, Stephen D. Roughley, Ben Davis, Helen Browne, Heather Simmonite, and Ben Gibbons
- Subjects
Male ,Pyrimidine ,Molecular model ,Stereochemistry ,Clinical Biochemistry ,Administration, Oral ,Pharmaceutical Science ,Biochemistry ,Mice ,Structure-Activity Relationship ,chemistry.chemical_compound ,Cell Line, Tumor ,Drug Discovery ,Animals ,Humans ,Pyrroles ,HSP90 Heat-Shock Proteins ,Surface plasmon resonance ,Molecular Biology ,Cell Proliferation ,Mice, Inbred BALB C ,Binding Sites ,biology ,Organic Chemistry ,Water ,Isothermal titration calorimetry ,HCT116 Cells ,Hsp90 ,In vitro ,Molecular Docking Simulation ,Pyrimidines ,chemistry ,Drug Design ,Chaperone (protein) ,biology.protein ,Molecular Medicine ,Fluorescence anisotropy ,Half-Life - Abstract
Inhibitors of the Hsp90 molecular chaperone are showing promise as anti-cancer agents. Here we describe a series of 4-aryl-5-cyanopyrrolo[2,3-d]pyrimidine ATP competitive Hsp90 inhibitors that were identified following structure-driven optimization of purine hits revealed by NMR based screening of a proprietary fragment library. Ligand-Hsp90 X-ray structures combined with molecular modeling led to the rational displacement of a conserved water molecule leading to enhanced affinity for Hsp90 as measured by fluorescence polarization, isothermal titration calorimetry and surface plasmon resonance assays. This displacement was achieved with a nitrile group, presenting an example of efficient gain in binding affinity with minimal increase in molecular weight. Some compounds in this chemical series inhibit the proliferation of human cancer cell lines in vitro and cause depletion of oncogenic Hsp90 client proteins and concomitant elevation of the co-chaperone Hsp70. In addition, one compound was demonstrated to be orally bioavailable in the mouse. This work demonstrates the power of structure-based design for the rapid evolution of potent Hsp90 inhibitors and the importance of considering conserved water molecules in drug design.
- Published
- 2012
10. Dynamic undocking and the quasi-bound state as tools for drug discovery
- Author
-
F. Javier Luque, Natalia Matassova, Xavier Barril, James Murray, Rod E. Hubbard, Peter Schmidtke, Ben Davis, Stephen D. Roughley, Sergio Ruiz-Carmona, and Lisa Baker
- Subjects
0301 basic medicine ,Virtual screening ,Molecular Structure ,Chemistry ,Drug discovery ,General Chemical Engineering ,General Chemistry ,Ligand (biochemistry) ,Ligands ,Small molecule ,Molecular Docking Simulation ,Combinatorial chemistry ,03 medical and health sciences ,030104 developmental biology ,Pharmaceutical Preparations ,Docking (molecular) ,Drug Discovery ,Hit rate ,Humans ,Thermodynamics ,Biochemical engineering ,HSP90 Heat-Shock Proteins ,Native contact - Abstract
There is a pressing need for new technologies that improve the efficacy and efficiency of drug discovery. Structure-based methods have contributed towards this goal but they focus on predicting the binding affinity of protein-ligand complexes, which is notoriously difficult. We adopt an alternative approach that evaluates structural, rather than thermodynamic, stability. As bioactive molecules present a static binding mode, we devised dynamic undocking (DUck), a fast computational method to calculate the work necessary to reach a quasi-bound state at which the ligand has just broken the most important native contact with the receptor. This non-equilibrium property is surprisingly effective in virtual screening because true ligands form more-resilient interactions than decoys. Notably, DUck is orthogonal to docking and other 'thermodynamic' methods. We demonstrate the potential of the docking-undocking combination in a fragment screening against the molecular chaperone and oncology target Hsp90, for which we obtain novel chemotypes and a hit rate that approaches 40%.
- Published
- 2016
11. A novel, small molecule inhibitor of Hsc70/Hsp70 potentiates Hsp90 inhibitor induced apoptosis in HCT116 colon carcinoma cells
- Author
-
Stephanie Geoffroy, Martin J. Drysdale, Pawel Dokurno, Zoe Daniels, James Brooke Murray, Paul Lavan, Geraint L. Francis, Natalia Matassova, Rachel Parsons, Douglas S. Williamson, Melanie Dopson, Terry Shaw, Yikang Wang, Antony Padfield, Mike Comer, Andrew Massey, Macias Alba, Michael Wood, Christopher John Graham, Helen Browne, and Vernalis (R&D) Ltd
- Subjects
Cancer Research ,Programmed cell death ,Breast Neoplasms ,Hsp90 ,Apoptosis ,Protein degradation ,Biology ,Toxicology ,Combination studies ,Hsp70 ,Hsp90 inhibitor ,Drug Delivery Systems ,Hsc70 ,Cell Line, Tumor ,Heat shock protein ,Antineoplastic Combined Chemotherapy Protocols ,Humans ,HSP70 Heat-Shock Proteins ,Pharmacokinetics ,Pharmacology (medical) ,HSP90 Heat-Shock Proteins ,ComputingMilieux_MISCELLANEOUS ,Cell Proliferation ,Caspase 7 ,Pharmacology ,Inhibitor of apoptosis domain ,Caspase 3 ,Cell growth ,HSC70 Heat-Shock Proteins ,Drug Synergism ,Purine Nucleosides ,Small molecule ,Oncology ,Colonic Neoplasms ,Immunology ,Cancer research ,Female ,Small molecule inhibitor - Abstract
The anti-apoptotic function of the 70 kDa family of heat shock proteins and their role in cancer is well documented. Dual targeting of Hsc70 and Hsp70 with siRNA induces proteasome-dependent degradation of Hsp90 client proteins and extensive tumor specific apoptosis as well as the potentiation of tumor cell apoptosis following pharmacological Hsp90 inhibition.We have previously described the discovery and synthesis of novel adenosine-derived inhibitors of the 70 kDa family of heat shock proteins; the first inhibitors described to target the ATPase binding domain. The in vitro activity of VER-155008 was evaluated in HCT116, HT29, BT474 and MDA-MB-468 carcinoma cell lines. Cell proliferation, cell apoptosis and caspase 3/7 activity was determined for VER-155008 in the absence or presence of small molecule Hsp90 inhibitors.VER-155008 inhibited the proliferation of human breast and colon cancer cell lines with GI(50)s in the range 5.3-14.4 microM, and induced Hsp90 client protein degradation in both HCT116 and BT474 cells. As a single agent, VER-155008 induced caspase-3/7 dependent apoptosis in BT474 cells and non-caspase dependent cell death in HCT116 cells. VER-155008 potentiated the apoptotic potential of a small molecule Hsp90 inhibitor in HCT116 but not HT29 or MDA-MB-468 cells. In vivo, VER-155008 demonstrated rapid metabolism and clearance, along with tumor levels below the predicted pharmacologically active level.These data suggest that small molecule inhibitors of Hsc70/Hsp70 phenotypically mimic the cellular mode of action of a small molecule Hsp90 inhibitor and can potentiate the apoptotic potential of a small molecule Hsp90 inhibitor in certain cell lines. The factors determining whether or not cells apoptose in response to Hsp90 inhibition or the combination of Hsp90 plus Hsc70/Hsp70 inhibition remain to be determined.
- Published
- 2009
12. Towards the discovery of drug-like RNA ligands?
- Author
-
Nicolas Foloppe, Fareed Aboul-ela, and Natalia Matassova
- Subjects
Models, Molecular ,Drug ,RNA, Untranslated ,media_common.quotation_subject ,Computational biology ,Biology ,Ligands ,Ribosome ,Structure-Activity Relationship ,Drug Discovery ,Humans ,Structure–activity relationship ,Binding site ,Protein Kinase Inhibitors ,media_common ,Antibacterial agent ,Pharmacology ,Binding Sites ,Molecular Structure ,RNA ,Ribosomal RNA ,Small molecule ,Anti-Bacterial Agents ,RNA, Bacterial ,Biochemistry ,RNA, Ribosomal ,Drug Design ,Computer-Aided Design ,Nucleic Acid Conformation ,RNA, Viral ,Ribosomes - Abstract
Targeting RNA with small molecule drugs is an area of great potential for therapeutic treatment of infections and possibly genetic and autoimmune diseases. However, a mature set of precedents and established methodology is lacking. The physicochemical properties of RNA raise specific issues and obstacles to development, and contribute to explain the distinct characteristics of natural RNA ligands, including antibiotics. Yet, RNA-targeting strategies are being implemented to reinvigorate antibacterial discovery by using the ribosomal X-ray structures to modify known antibiotics. To exploit further these structures, we suggest the use of existing protein kinase-directed libraries of drug-like compounds to target the A-site of the bacterial ribosome, on the basis of a specific structural hypothesis.
- Published
- 2006
13. Structural Basis for Contrasting Activities of Ribosome Binding Thiazole Antibiotics
- Author
-
Fareed Aboul-ela, Georg Lentzen, Alastair I.H. Murchie, Natalia Matassova, and Roscoe Klinck
- Subjects
Models, Molecular ,Magnetic Resonance Spectroscopy ,Molecular Sequence Data ,Clinical Biochemistry ,Biology ,Methylation ,Biochemistry ,Ribosome ,Thiostrepton ,chemistry.chemical_compound ,Bacteriocins ,23S ribosomal RNA ,Drug Discovery ,Molecular Biology ,Pharmacology ,Binding Sites ,Base Sequence ,Hydrolysis ,RNA ,Methyltransferases ,General Medicine ,Ribosomal RNA ,Anti-Bacterial Agents ,Elongation factor ,RNA, Ribosomal, 23S ,Thiazoles ,chemistry ,Molecular Medicine ,Guanosine Triphosphate ,Peptides ,Ribosomes ,Nosiheptide ,Protein Binding ,Binding domain - Abstract
Thiostrepton and micrococcin inhibit protein synthesis by binding to the L11 binding domain (L11BD) of 23S ribosomal RNA. The two compounds are structurally related, yet they produce different effects on ribosomal RNA in footprinting experiments and on elongation factor-G (EF-G)-dependent GTP hydrolysis. Using NMR and an assay based on A1067 methylation by thiostrepton-resistance methyltransferase, we show that the related thiazoles, nosiheptide and siomycin, also bind to this region. The effect of all four antibiotics on EF-G-dependent GTP hydrolysis and EF-G-GDP-ribosome complex formation was studied. Our NMR and biochemical data demonstrate that thiostrepton, nosiheptide, and siomycin share a common profile, which differs from that of micrococcin. We have generated a three-dimensional (3D) model for the interaction of thiostrepton with L11BD RNA. The model rationalizes the differences between micrococcin and the thiostrepton-like antibiotics interacting with L11BD.
- Published
- 2003
14. The bacterial ribosome, a promising focus for structure-based drug design
- Author
-
David Justin Charles Knowles, Natalia Matassova, Alastair I H Murchie, and Nicolas Foloppe
- Subjects
Pharmacology ,Drug ,Molecular interactions ,Virtual screening ,Bacteria ,medicine.drug_class ,media_common.quotation_subject ,Bacterial ribosome ,Antibiotics ,Computational Biology ,Drug design ,Computational biology ,Biology ,Ribosome ,Combinatorial chemistry ,Anti-Bacterial Agents ,Drug Design ,Drug Discovery ,medicine ,Structure based ,Ribosomes ,media_common - Abstract
Recent crystal structures of the bacterial ribosome have identified the complex molecular interactions involved in antibiotic-ribosome recognition. Insights into the binding of aminoglycosides, macrolides, tetracyclines and other antibiotics provide opportunities for computational, structure-based approaches to be used in the design of appropriate modifications to existing antibiotics as well as in the discovery of completely new drug classes.
- Published
- 2002
15. Off-rate screening (ORS) by surface plasmon resonance. An efficient method to kinetically sample hit to lead chemical space from unpurified reaction products
- Author
-
James Brooke Murray, Stephen D. Roughley, Paul Brough, and Natalia Matassova
- Subjects
Empirical data ,Drug discovery ,Chemistry ,Drug Evaluation, Preclinical ,Hit to lead ,Biosensing Techniques ,Surface Plasmon Resonance ,Off rate ,Combinatorial chemistry ,Chemical space ,Small Molecule Libraries ,Dissociation rate constant ,Kinetics ,Pharmaceutical Preparations ,Phase (matter) ,Drug Discovery ,Molecular Medicine ,Humans ,Surface plasmon resonance - Abstract
The dissociation rate constant kd (off-rate) is the component of ligand–protein binding with the most significant potential to enhance compound potency. Here we provide theoretical and empirical data to show that this parameter can be determined accurately from unpurified reaction products containing designed test compounds. This screening protocol is amenable to parallel chemistry, provides efficiencies of time and materials, and complements existing methodologies for the hit-to-lead phase in fragment-based drug discovery.
- Published
- 2014
16. Fatty acid amide hydrolase inhibitors. 3: tetra-substituted azetidine ureas with in vivo activity
- Author
-
Hart Terance William, Rachel Parsons, Ben Gibbons, Helen Browne, Macias Alba, Heather Simmonite, Sarah Dugdale, Sean Lightowler, Howard Langh Am Mansell, Guy A. Kennett, Anthony Padfield, Geraint L. Francis, Robert M. Pratt, Teresa Brooks, Timothy Haymes, Jalanie D’Alessandro, Natalia Matassova, K Benwell, Melanie Wong, Steven B. Walls, Stephen D. Roughley, A Misra, Zoe Daniels, Angela Merrett, Alan D. Robertson, and Kiri Tan
- Subjects
Models, Molecular ,ERG1 Potassium Channel ,Stereochemistry ,Clinical Biochemistry ,Azetidine ,Pharmaceutical Science ,Stereoisomerism ,Biochemistry ,Amidohydrolases ,chemistry.chemical_compound ,Structure-Activity Relationship ,Cytochrome P-450 Enzyme System ,Fatty acid amide hydrolase ,In vivo ,Drug Discovery ,Structure–activity relationship ,Animals ,Cytochrome P-450 Enzyme Inhibitors ,Humans ,Urea ,Enzyme Inhibitors ,Molecular Biology ,Dose-Response Relationship, Drug ,Molecular Structure ,Organic Chemistry ,Anandamide ,Ether-A-Go-Go Potassium Channels ,Rats ,nervous system ,chemistry ,Molecular Medicine ,Azetidines ,lipids (amino acids, peptides, and proteins) ,psychological phenomena and processes - Abstract
We describe here our attempts to optimise the human fatty acid amide hydrolase (FAAH) inhibition and physicochemical properties of our previously reported tetrasubstituted azetidine urea FAAH inhibitor, VER-156084. We describe the SAR of a series of analogues and conclude with the demonstration of in vivo dose-dependant FAAH inhibition in an anandamide-loading study in rats.
- Published
- 2011
17. Adenosine-derived inhibitors of 78 kDa glucose regulated protein (Grp78) ATPase: insights into isoform selectivity
- Author
-
Nicola Allen, Lindsey Terry, Andrew Massey, Michael Wood, Natalia Matassova, Allan E. Surgenor, Terry Shaw, Martin J. Drysdale, Geraint L. Francis, Yikang Wang, Pawel Dokurno, Christopher John Graham, Rachel Parsons, Rob Howes, Jenifer Borgognoni, Douglas S. Williamson, Zoe Daniels, James Brooke Murray, Macias Alba, and Alexandra Clay
- Subjects
Models, Molecular ,Protein Conformation ,Adenylyl Imidodiphosphate ,Calorimetry ,Crystallography, X-Ray ,Ligands ,chemistry.chemical_compound ,Structure-Activity Relationship ,Adenosine Triphosphate ,Drug Discovery ,Protein A/G ,Protein Isoforms ,HSP70 Heat-Shock Proteins ,Binding site ,Furans ,Endoplasmic Reticulum Chaperone BiP ,Heat-Shock Proteins ,Adenosine Triphosphatases ,Binding Sites ,biology ,Binding protein ,HSC70 Heat-Shock Proteins ,Isothermal titration calorimetry ,Stereoisomerism ,Surface Plasmon Resonance ,78 kDa Glucose-Regulated Protein ,chemistry ,Biochemistry ,Purines ,Chaperone (protein) ,biology.protein ,Molecular Medicine ,Thermodynamics ,Protein G ,Adenosine triphosphate ,Protein Binding - Abstract
78 kDa glucose-regulated protein (Grp78) is a heat shock protein (HSP) involved in protein folding that plays a role in cancer cell proliferation. Binding of adenosine-derived inhibitors to Grp78 was characterized by surface plasmon resonance and isothermal titration calorimetry. The most potent compounds were 13 (VER-155008) with K(D) = 80 nM and 14 with K(D) = 60 nM. X-ray crystal structures of Grp78 bound to ATP, ADPnP, and adenosine derivative 10 revealed differences in the binding site between Grp78 and homologous proteins.
- Published
- 2011
18. Fatty acid amide hydrolase inhibitors. Surprising selectivity of chiral azetidine ureas
- Author
-
Alan Robertson, Guy A. Kennett, Stephen D. Roughley, Geraint L. Francis, K Benwell, Melanie Wong, Rachel Parsons, A Misra, Timothy Haymes, Hart Terance William, Howard Langh Am Mansell, Macias Alba, Anthony Padfield, Sean Lightowler, Jalanie D’Alessandro, Natalia Matassova, Ben Gibbons, Teresa Brooks, Steven B. Walls, Robert M. Pratt, and Pawel Dokurno
- Subjects
Stereochemistry ,Chemistry, Pharmaceutical ,Clinical Biochemistry ,Azetidine ,Pharmaceutical Science ,Ether ,Stereoisomerism ,Biochemistry ,Catalysis ,Amidase ,Amidohydrolases ,chemistry.chemical_compound ,Mice ,Piperidines ,Receptor, Cannabinoid, CB1 ,Fatty acid amide hydrolase ,Drug Discovery ,Animals ,Humans ,Urea ,Enzyme Inhibitors ,Molecular Biology ,chemistry.chemical_classification ,Mice, Knockout ,biology ,Organic Chemistry ,Rats ,Enzyme ,nervous system ,chemistry ,Models, Chemical ,Enzyme inhibitor ,Drug Design ,biology.protein ,Molecular Medicine ,Azetidines ,lipids (amino acids, peptides, and proteins) ,Selectivity ,psychological phenomena and processes - Abstract
We report the discovery of a novel, chiral azetidine urea inhibitor of Fatty Acid Amide Hydrolase (FAAH,) and describe the surprising species selectivity of VER-156084 versus rat and human FAAH and also hCB1.
- Published
- 2009
19. RNA as a drug target
- Author
-
Martin J, Drysdale, Georg, Lentzen, Natalia, Matassova, Alastair I H, Murchie, Fareed, Aboul-Ela, and Mohammad, Afshar
- Subjects
Protein Synthesis Inhibitors ,RNA, Bacterial ,Structure-Activity Relationship ,Anti-HIV Agents ,RNA, Ribosomal ,Drug Design ,Animals ,Humans ,Nucleic Acid Conformation ,RNA, Viral ,Ribosomes ,Anti-Bacterial Agents - Abstract
In the antiviral and antibacterial area, increasing drug resistance means that there is an ever growing need for novel approaches towards structures and mechanisms which avoid the current problems. The huge increase in high resolution structural data is set to make a dramatic impact on targeting RNA as a drug target. The examples of the RNA binding antibiotics, particularly, the totally synthetic oxazolidinones, should help persuade the skceptics that clinically useful, selective drugs can be obtained from targeting RNA directly.
- Published
- 2003
20. 2 RNA as a Drug Target
- Author
-
Alastair I.H. Murchie, Martin J. Drysdale, Natalia Matassova, Mohammad Afshar, Georg Lentzen, and Fareed Aboul-ela
- Subjects
biology ,medicine.drug_class ,Drug target ,Antibiotics ,RNA ,Drug resistance ,Computational biology ,Pharmacology ,biology.organism_classification ,Ribosome ,Transcription (biology) ,Lentivirus ,medicine ,Antibacterial agent - Abstract
In the antiviral and antibacterial area, increasing drug resistance means that there is an ever growing need for novel approaches towards structures and mechanisms which avoid the current problems. The huge increase in high resolution structural data is set to make a dramatic impact on targeting RNA as a drug target. The examples of the RNA binding antibiotics, particularly, the totally synthetic oxazolidinones, should help persuade the skceptics that clinically useful, selective drugs can be obtained from targeting RNA directly.
- Published
- 2002
21. Abstract 2782: Bcl-2 selective antagonists show antitumor activity without dose limiting platelet toxicity
- Author
-
Claire L. Nunns, Francisco Cruzalegui, Starck Jérôme-Benoît, John A. Hickman, Michael Wood, Patrick Casara, Stéphane Depil, James Edward Paul Davidson, Natalia Matassova, Christopher John Graham, Alain Bruno, Pawel Dokurno, Thierry Le Diguarher, Alain Pierre, James Murray, Neil Whitehead, Guillaume De Nanteuil, Olivier Geneste, and Chen I-Jen
- Subjects
Cancer Research ,Programmed cell death ,Pharmacology ,Biology ,Oncology ,Mechanism of action ,Apoptosis ,In vivo ,Cell culture ,Toxicity ,medicine ,Platelet ,medicine.symptom ,IC50 - Abstract
Proteins of the Bcl-2 family are central regulators of programmed cell death. Pro-survival Bcl-2 proteins, such as Bcl-2, Bcl-xL and Mcl-1 are often over-expressed in human tumours and participate in tumour initiation, progression and chemo-resistance. Therefore drugs targeting these pro-survival Bcl-2 proteins represent a promising therapeutic approach for cancer treatment. The most advanced drug targeting this protein family is ABT-263, a potent Bcl-2 and Bcl-xL inhibitor showing anti-tumour efficacy in preclinical models of leukaemia and small cell lung carcinoma. Survival of circulating platelets has been shown to be highly Bcl-xL dependent; consequently the dose-limiting toxicity of ABT-263 is an on-target peripheral thrombocytopenia. We have used a range of biophysical methods to guide the structure-based generation of a significant number of small molecules,* which bind with high affinity (MW < 780; KD < 1 nM) to the BH3 binding groove of Bcl-2, and with high selectivity versus other members of the Bcl-2 family (selectivity > 100 fold). In cellular assays, our lead compounds efficiently displace Bax from Bcl-2 with near complete inhibition of Bcl-2 / Bax co-immunoprecipitation at 100 nM. These compounds are strong inducers of cell death in Bcl-2 dependent cellular models such as the acute myeloblastic leukaemia (AML) cell line RS4;11, affording sub-10 nM IC50's for the most potent compounds. In vivo, in agreement with their mechanism of action, these Bcl-2 selective inhibitors, given either intravenously or orally, elicit a rapid (30 min iv, and 2 hours po) and strong apoptotic response in mouse xenografts of the RS4:11 cell line. When the most potent compounds are given orally to RS4;11 xenograft-bearing mice, apoptosis in tumor cells is induced more than 15 fold (at 25 mg/kg) and more than 20 fold (at 50 mg/kg) compared to untreated mice. Importantly, in agreement with the selectivity of the compounds for Bcl-2 versus Bcl-xL, no platelet loss was observed in mice treated with our compounds, in sharp contrast to ABT-263. Finally, we observe very robust anti-tumor activity when a lead compound is given orally at 50 mg/kg and 100 mg/kg (with complete regression at 100 mg/kg) in an RS4;11 mouse xenograft model. This anti-tumor activity was similar whether the compound was dosed daily or twice a week over two weeks. Altogether our data demonstrate that highly Bcl-2 selective antagonists show anti-tumor activity and no platelet toxicity, in contrast to Bcl-2 / Bcl-xL dual inhibitors. Such compounds represent promising drug candidates for the treatment of Bcl-2 dependent malignancies such as chronic lymphocytic leukaemia (CLL) and other leukaemias and lymphomas. * Chemical structures of compounds will not be disclosed. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2782. doi:1538-7445.AM2012-2782
- Published
- 2012
22. Corrigendum to 'Structure-guided design of α-amino acid-derived Pin1 inhibitors' [Bioorg. Med. Chem. Lett. 20 (2010) 586]
- Author
-
Claire L. Nunns, Jonathan D. Moore, Simon F. Scrace, David A. Robinson, Lisa Baker, Pawel Dokurno, James Brooke Murray, Natalia Matassova, Louisa Gueritz, Andrew J. Potter, Stuart C. Ray, Christine M. Richardson, Ben Davis, Christopher J. Bryant, and Allan E. Surgenor
- Subjects
chemistry.chemical_classification ,chemistry ,Stereochemistry ,Organic Chemistry ,Clinical Biochemistry ,Drug Discovery ,PIN1 ,Pharmaceutical Science ,Molecular Medicine ,Molecular Biology ,Biochemistry ,Amino acid - Published
- 2010
23. Abstract A212: A novel, small molecule inhibitor of Hsc70/Hsp70 potentiates Hsp90 inhibitor-induced apoptosis in HCT116 colon carcinoma cells
- Author
-
Andrew Massey, Michael Wood, Rachel Parsons, Pawel Dokurno, Yikang Wang, Zoe Daniels, Christopher John Graham, Terry Shaw, Macias Alba, Jennifer Borgognoni, Geraint L. Francis, Natalia Matassova, Douglas S. Williamson, Helen Browne, Martin J. Drysdale, and James Brooke Murray
- Subjects
Cancer Research ,Programmed cell death ,Oncology ,Apoptosis ,Cell growth ,biology.protein ,Cell cycle ,Biology ,Protein degradation ,Hsp90 ,Small molecule ,Hsp90 inhibitor ,Cell biology - Abstract
The role of the 70 kDa heat shock protein isoforms (Hsc70 and Hsp70) in cancer development and progression through their ability to inhibit apoptosis and via their role as Hsp90 co-chaperones has been well documented. Dual targeting of Hsc70 and Hsp70 with siRNA has previously been demonstrated to induce proteasome-dependent degradation of Hsp90 client proteins and extensive tumor specific apoptosis as well as the potentiation of tumor cell apoptosis following pharmacological Hsp90 inhibition. The design and synthesis of novel adenosine-derived inhibitors of Hsp70, guided by modelling and X-ray crystallographic structures of these compounds in complex with Hsc70/BAG-1, has been described.1 These were the first inhibitors described to target the ATPase binding domain of this family of chaperones. Many of these compounds exhibited submicromolar affinity for Hsp70, were highly selective over Hsp90, and displayed in vitro activity against a variety of human tumor cell lines. We further describe the in vitro mode of action of one of the most potent analogues, VER-155008 in HCT116, HT29, BT474 and MDA-MB-468 carcinoma cell lines. Cell proliferation, cell cycle, cell apoptosis and caspase 3/7 activity was determined for VER-155008 in the absence or presence of small molecule Hsp90 inhibitors. VER-155008 inhibited the proliferation of human breast and colon cancer cell lines with GI50s in the range 5.3 to 14.4 M, and induced Hsp90 client protein degradation in both HCT116 and BT474 cells. As a single agent, VER-155008 induced caspase-3/7 dependent apoptosis in BT474 cells and non-caspase dependent cell death in HCT116 and HT29 cells. VER-155008 potentiated the apoptotic potential of the small molecule Hsp90 inhibitors VER-821602 and 17-AAG in HCT116 but not HT29 or MDA-MB-468 cells. In vivo, VER-155008 demonstrated rapid metabolism and clearance, along with tumor levels below the predicted pharmacologically active level. These data suggest that as a single agent, small molecule inhibitors of Hsc70/Hsp70 phenotypically mimic the cellular mode of action of a small molecule Hsp90 inhibitor and can potentiate the apoptotic potential of a small molecule Hsp90 inhibitor in certain cell lines. The factors determining whether or not cells apoptose in response to Hsp90 inhibition or the combination of Hsp90 plus Hsc70/Hsp70 inhibition remain to be determined. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):A212.
- Published
- 2009
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.