I03-Macromolecular Crystallography
I04-1-Macromolecular Crystallography (fixed wavelength)
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Olivier A.
Pierrat
,
Manjuan
Liu
,
Gavin W.
Collie
,
Kartika N.
Shetty
,
Matthew J.
Rodrigues
,
Yann-Vai
Le Bihan
,
Emma A.
Gunnell
,
P. Craig
Mcandrew
,
Mark
Stubbs
,
Martin G.
Rowlands
,
Norhakim
Yahya
,
Erald
Shehu
,
Rachel
Talbot
,
Lisa
Pickard
,
Benjamin R.
Bellenie
,
Kwai-Ming J.
Cheung
,
Ludovic
Drouin
,
Paolo
Innocenti
,
Hannah
Woodward
,
Owen A.
Davis
,
Matthew G.
Lloyd
,
Ana
Varela
,
Rosemary
Huckvale
,
Fabio
Broccatelli
,
Michael
Carter
,
David
Galiwango
,
Angela
Hayes
,
Florence I.
Raynaud
,
Christopher
Bryant
,
Steven
Whittaker
,
Olivia W.
Rossanese
,
Swen
Hoelder
,
Rosemary
Burke
,
Rob L. M.
Van Montfort
Open Access
Abstract: By suppressing gene transcription through the recruitment of corepressor proteins, B-cell lymphoma 6 (BCL6) protein controls a transcriptional network required for the formation and maintenance of B-cell germinal centres. As BCL6 deregulation is implicated in the development of Diffuse Large B-Cell Lymphoma, we sought to discover novel small molecule inhibitors that disrupt the BCL6-corepressor protein–protein interaction (PPI). Here we report our hit finding and compound optimisation strategies, which provide insight into the multi-faceted orthogonal approaches that are needed to tackle this challenging PPI with small molecule inhibitors. Using a 1536-well plate fluorescence polarisation high throughput screen we identified multiple hit series, which were followed up by hit confirmation using a thermal shift assay, surface plasmon resonance and ligand-observed NMR. We determined X-ray structures of BCL6 bound to compounds from nine different series, enabling a structure-based drug design approach to improve their weak biochemical potency. We developed a time-resolved fluorescence energy transfer biochemical assay and a nano bioluminescence resonance energy transfer cellular assay to monitor cellular activity during compound optimisation. This workflow led to the discovery of novel inhibitors with respective biochemical and cellular potencies (IC50s) in the sub-micromolar and low micromolar range.
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Nov 2022
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I03-Macromolecular Crystallography
I04-1-Macromolecular Crystallography (fixed wavelength)
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Owen A.
Davis
,
Kwai-Ming J.
Cheung
,
Alfie
Brennan
,
Matthew G.
Lloyd
,
Matthew J.
Rodrigues
,
Olivier A.
Pierrat
,
Gavin W.
Collie
,
Yann-Vai
Le Bihan
,
Rosemary
Huckvale
,
Alice C.
Harnden
,
Ana
Varela
,
Michael D.
Bright
,
Paul
Eve
,
Angela
Hayes
,
Alan T.
Henley
,
Michael D.
Carter
,
P. Craig
Mcandrew
,
Rachel
Talbot
,
Rosemary
Burke
,
Rob
Van Montfort
,
Florence I.
Raynaud
,
Olivia W.
Rossanese
,
Mirco
Meniconi
,
Benjamin R.
Bellenie
,
Swen
Hoelder
Open Access
Abstract: To identify new chemical series with enhanced binding affinity to the BTB domain of B-cell lymphoma 6 protein, we targeted a subpocket adjacent to Val18. With no opportunities for strong polar interactions, we focused on attaining close shape complementarity by ring fusion onto our quinolinone lead series. Following exploration of different sized rings, we identified a conformationally restricted core which optimally filled the available space, leading to potent BCL6 inhibitors. Through X-ray structure-guided design, combined with efficient synthetic chemistry to make the resulting novel core structures, a >300-fold improvement in activity was obtained by the addition of seven heavy atoms.
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Jun 2022
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I03-Macromolecular Crystallography
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Rosemary
Huckvale
,
Alice C.
Harnden
,
Kwai-Ming J.
Cheung
,
Olivier A.
Pierrat
,
Rachel
Talbot
,
Gary M.
Box
,
Alan T.
Henley
,
Alexis K.
De Haven Brandon
,
Albert E.
Hallsworth
,
Michael D.
Bright
,
Hafize Aysin
Akpinar
,
Daniel S. J.
Miller
,
Dalia
Tarantino
,
Sharon
Gowan
,
Angela
Hayes
,
Emma A.
Gunnell
,
Alfie
Brennan
,
Owen A.
Davis
,
Louise D.
Johnson
,
Selby
De Klerk
,
Craig
Mcandrew
,
Yann-Vai
Le Bihan
,
Mirco
Meniconi
,
Rosemary
Burke
,
Vladimir
Kirkin
,
Rob L. M.
Van Montfort
,
Florence I.
Raynaud
,
Olivia W.
Rossanese
,
Benjamin R.
Bellenie
,
Swen
Hoelder
Open Access
Abstract: The transcriptional repressor BCL6 is an oncogenic driver found to be deregulated in lymphoid malignancies. Herein, we report the optimization of our previously reported benzimidazolone molecular glue-type degrader CCT369260 to CCT373566, a highly potent probe suitable for sustained depletion of BCL6 in vivo. We observed a sharp degradation SAR, where subtle structural changes conveyed the ability to induce degradation of BCL6. CCT373566 showed modest in vivo efficacy in a lymphoma xenograft mouse model following oral dosing.
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Jun 2022
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B21-High Throughput SAXS
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Ami
Miller
,
Adam
Leach
,
Jemima
Thomas
,
Craig
Mcandrew
,
Emma
Bentley
,
Giada
Mattiuzzo
,
Lijo
John
,
Ali
Mirazimi
,
Gemma
Harris
,
Nadisha
Gamage
,
Stephen
Carr
,
Hanif
Ali
,
Rob
Van Montfort
,
Terence
Rabbitts
Diamond Proposal Number(s):
[27159]
Open Access
Abstract: Approaches are needed for therapy of the severe acute respiratory syndrome from SARS-CoV-2 coronavirus (COVID-19). Interfering with the interaction of viral antigens with the angiotensin converting enzyme 2 (ACE-2) receptor is a promising strategy by blocking the infection of the coronaviruses into human cells. We have implemented a novel protein engineering technology to produce a super-potent tetravalent form of ACE2, coupled to the human immunoglobulin γ1 Fc region, using a self-assembling, tetramerization domain from p53 protein. This high molecular weight Quad protein (ACE2-Fc-TD) retains binding to the SARS-CoV-2 receptor binding spike protein and can form a complex with the spike protein plus anti-viral antibodies. The ACE2-Fc-TD acts as a powerful decoy protein that out-performs soluble monomeric and dimeric ACE2 proteins and blocks both SARS-CoV-2 pseudovirus and SARS-CoV-2 virus infection with greatly enhanced efficacy. The ACE2 tetrameric protein complex promise to be important for development as decoy therapeutic proteins against COVID-19. In contrast to monoclonal antibodies, ACE2 decoy is unlikely to be affected by mutations in SARS-CoV-2 that are beginning to appear in variant forms. In addition, ACE2 multimeric proteins will be available as therapeutic proteins should new coronaviruses appear in the future because these are likely to interact with ACE2 receptor.
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May 2021
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I03-Macromolecular Crystallography
I04-1-Macromolecular Crystallography (fixed wavelength)
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Benjamin R.
Bellenie
,
Kwai-Ming J.
Cheung
,
Ana
Varela
,
Olivier A.
Pierrat
,
Gavin W.
Collie
,
Gary M.
Box
,
Michael D.
Bright
,
Sharon
Gowan
,
Angela
Hayes
,
Matthew J.
Rodrigues
,
Kartika N.
Shetty
,
Michael
Carter
,
Owen A.
Davis
,
Alan T.
Henley
,
Paolo
Innocenti
,
Louise D.
Johnson
,
Manjuan
Liu
,
Selby
De Klerk
,
Yann-Vai
Le Bihan
,
Matthew G.
Lloyd
,
P. Craig
Mcandrew
,
Erald
Shehu
,
Rachel
Talbot
,
Hannah L.
Woodward
,
Rosemary
Burke
,
Vladimir
Kirkin
,
Rob L. M.
Van Montfort
,
Florence I.
Raynaud
,
Olivia W.
Rossanese
,
Swen
Hoelder
Diamond Proposal Number(s):
[14891, 20145]
Open Access
Abstract: Deregulation of the transcriptional repressor BCL6 enables tumorigenesis of germinal center B-cells, and hence BCL6 has been proposed as a therapeutic target for the treatment of diffuse large B-cell lymphoma (DLBCL). Herein we report the discovery of a series of benzimidazolone inhibitors of the protein-protein interaction between BCL6 and its co-repressors. A subset of these inhibitors were found to cause rapid degradation of BCL6, and optimization of pharmacokinetic properties led to the discovery of 5-((5-chloro-2-((3R,5S)-4,4-difluoro-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-3-(3-hydroxy-3-methylbutyl)-1-methyl-1,3-dihydro-2H-benzo[d]imidazol-2-one (CCT369260), which reduces BCL6 levels in a lymphoma xenograft mouse model following oral dosing.
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Apr 2020
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I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
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Open Access
Abstract: Screening a 3-aminopyridin-2-one based fragment library against a 26-kinase panel representative of the human kinome identified 3-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-2(1H)-one (2) and 3-amino-5-(pyridin-4-yl)pyridin-2(1H)-one (3) as ligand efficient inhibitors of the mitotic kinase Monopolar Spindle 1 (MPS1) and the Aurora kinase family. These kinases are well recognised as attractive targets for therapeutic intervention for treating cancer. Elucidation of the binding mode of these fragments and their analogues has been carried out by X-ray crystallography. Structural studies have identified key interactions with a conserved lysine residue and have highlighted potential regions of MPS1 which could be targeted to improve activity and selectivity.
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Apr 2018
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I04-Macromolecular Crystallography
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Diamond Proposal Number(s):
[14891, 6385, 8015, 10088]
Open Access
Abstract: TLE1 is an oncogenic transcriptional co-repressor that exerts its repressive effects through binding of transcription factors. Inhibition of this protein-protein interaction represents a putative cancer target but no small molecule inhibitors have been published for this challenging interface. In this manuscript, we report the structure enabled design and synthesis of a constrained peptide inhibitor of TLE1. Our design featured introduction of a four carbon atom linker into the peptide epitope found in many TLE1 binding partners. We developed a concise synthetic route to a proof of concept peptide cycFWRPW. Biophysical testing by ITC and thermal shift assays showed that whilst the constrained peptide bound potently, it had an approximately five fold higher Kd than the unconstrained peptide. Our co-crystal structure suggested that the reduced affinity is likely due to a small shift of one side-chain compared to the otherwise well conserved conformation of the acyclic peptide. Our work describes a constrained peptide inhibitor that may serve as the basis for improved inhibitors.
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Mar 2017
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I04-1-Macromolecular Crystallography (fixed wavelength)
I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
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Paolo
Innocenti
,
Hannah L.
Woodward
,
Savade
Solanki
,
Sébastien
Naud
,
Isaac
Westwood
,
Nora
Cronin
,
Angela
Hayes
,
Jennie
Roberts
,
Alan T.
Henley
,
Ross
Baker
,
Amir
Faisal
,
Grace Wing-Yan
Mak
,
Gary
Box
,
Melanie
Valenti
,
Alexis
De Haven Brandon
,
Lisa
O’ Fee
,
Harry
Saville
,
Jessica
Schmitt
,
Berry
Matijssen
,
Rosemary
Burke
,
Rob
Van Montfort
,
Florence I.
Raynaud
,
Suzanne A.
Eccles
,
Spiros
Linardopoulos
,
Julian
Blagg
,
Swen
Hoelder
Open Access
Abstract: Monopolar spindle 1 (MPS1) plays a central role in the transition of cells from metaphase to anaphase and is one of the main components of the spindle assembly checkpoint. Chromosomally unstable cancer cells rely heavily on MPS1 to cope with the stress arising from abnormal numbers of chromosomes and centrosomes and are thus more sensitive to MPS1 inhibition than normal cells. We report the discovery and optimization of a series of new pyrido[3,4-d]pyrimidine based inhibitors via a structure-based hybridization approach from our previously reported inhibitor CCT251455 and a modestly potent screening hit. Compounds in this novel series display excellent potency and selectivity for MPS1, which translates into biomarker modulation in an in vivo human tumor xenograft model.
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Apr 2016
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I02-Macromolecular Crystallography
I04-1-Macromolecular Crystallography (fixed wavelength)
I04-Macromolecular Crystallography
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M. D.
Gurden
,
Isaac
Westwood
,
A.
Faisal
,
S.
Naud
,
K. M. J.
Cheung
,
C.
Mcandrew
,
A.
Wood
,
J.
Schmitt
,
K.
Boxall
,
G.
Mak
,
P.
Workman
,
R.
Burke
,
S.
Hoelder
,
J.
Blagg
,
Rob
Van Montfort
,
S.
Linardopoulos
Diamond Proposal Number(s):
[8015, 10088]
Abstract: Acquired resistance to therapy is perhaps the greatest challenge to effective clinical management of cancer. With several inhibitors of the mitotic checkpoint kinase MPS1 in preclinical development, we sought to investigate how resistance against these inhibitors may arise so that mitigation or bypass strategies could be addressed as early as possible. Toward this end, we modeled acquired resistance to the MPS1 inhibitors AZ3146, NMS-P715, and CCT251455, identifying five point mutations in the kinase domain of MPS1 that confer resistance against multiple inhibitors. Structural studies showed how the MPS1 mutants conferred resistance by causing steric hindrance to inhibitor binding. Notably, we show that these mutations occur in nontreated cancer cell lines and primary tumor specimens, and that they also preexist in normal lymphoblast and breast tissues. In a parallel piece of work, we also show that the EGFR p.T790M mutation, the most common mutation conferring resistance to the EGFR inhibitor gefitinib, also preexists in cancer cells and normal tissue. Our results therefore suggest that mutations conferring resistance to targeted therapy occur naturally in normal and malignant cells and these mutations do not arise as a result of the increased mutagenic plasticity of cancer cells.
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Aug 2015
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