Catherine A. Kettleborough
Medical Research Council
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Featured researches published by Catherine A. Kettleborough.
Cell | 2007
Sharon Yeoh; Rebecca A. O'donnell; Konstantinos Koussis; Anton R. Dluzewski; Keith H. Ansell; Simon A. Osborne; Fiona Hackett; Chrislaine Withers-Martinez; G. H. Mitchell; L. H. Bannister; Justin S. Bryans; Catherine A. Kettleborough; Michael J. Blackman
The most virulent form of malaria is caused by waves of replication of blood stages of the protozoan pathogen Plasmodium falciparum. The parasite divides within an intraerythrocytic parasitophorous vacuole until rupture of the vacuole and host-cell membranes releases merozoites that invade fresh erythrocytes to repeat the cycle. Despite the importance of merozoite egress for disease progression, none of the molecular factors involved are known. We report that, just prior to egress, an essential serine protease called PfSUB1 is discharged from previously unrecognized parasite organelles (termed exonemes) into the parasitophorous vacuole space. There, PfSUB1 mediates the proteolytic maturation of at least two essential members of another enzyme family called SERA. Pharmacological blockade of PfSUB1 inhibits egress and ablates the invasive capacity of released merozoites. Our findings reveal the presence in the malarial parasitophorous vacuole of a regulated, PfSUB1-mediated proteolytic processing event required for release of viable parasites from the host erythrocyte.
Journal of Biomolecular Screening | 2013
Craig Southern; Jennifer M. Cook; Zaynab Neetoo-Isseljee; Debra Taylor; Catherine A. Kettleborough; Andy Merritt; Daniel Bassoni; William Raab; Elizabeth Quinn; Tom Wehrman; Anthony P. Davenport; Andrew J. Brown; Andrew Green; Mark J. Wigglesworth; Steve Rees
A variety of G-protein–coupled receptor (GPCR) screening technologies have successfully partnered a number of GPCRs with their cognate ligands. GPCR-mediated β-arrestin recruitment is now recognized as a distinct intracellular signaling pathway, and ligand-receptor interactions may show a bias toward β-arrestin over classical GPCR signaling pathways. We hypothesized that the failure to identify native ligands for the remaining orphan GPCRs may be a consequence of biased β-arrestin signaling. To investigate this, we assembled 10 500 candidate ligands and screened 82 GPCRs using PathHunter β-arrestin recruitment technology. High-quality screening assays were validated by the inclusion of liganded receptors and the detection and confirmation of these established ligand-receptor pairings. We describe a candidate endogenous orphan GPCR ligand and a number of novel surrogate ligands. However, for the majority of orphan receptors studied, measurement of β-arrestin recruitment did not lead to the identification of cognate ligands from our screening sets. β-Arrestin recruitment represents a robust GPCR screening technology, and ligand-biased signaling is emerging as a therapeutically exploitable feature of GPCR biology. The identification of cognate ligands for the orphan GPCRs and the extent to which receptors may exist to preferentially signal through β-arrestin in response to their native ligand remain to be determined.
Tuberculosis | 2011
Kathryn E.A. Lougheed; Simon A. Osborne; Barbara Saxty; David Whalley; Tim Chapman; Nathalie Bouloc; Jasveen Chugh; Timothy J. Nott; Dony Patel; Vicky L. Spivey; Catherine A. Kettleborough; Justin S. Bryans; Debra L. Taylor; Stephen J. Smerdon; Roger S. Buxton
Summary PknB is an essential serine/threonine kinase of Mycobacterium tuberculosis with possible roles in a number of signalling pathways involved in cell division and metabolism. We screened a library of >50,000 compounds for inhibitors of the in vitro phosphorylation of GarA (Rv1827) by PknB and identified a number of inhibitors. A program of synthetic medicinal chemistry was subsequently conducted around one class of inhibitors and was successful in generating ATP competitive inhibitors with potency in the nanomolar range. Compounds in this class showed cross-reactivity with the related M. tuberculosis kinase, PknF, but not with PknG in an in vitro autophosphorylation assay. These synthesised inhibitors were able to prevent the growth of M. tuberculosis in an Alamar blue assay and in an intracellular model of infection, but only in the micromolar range. We attempted to determine if cell wall permeability was an explanation for the discrepancy between the potent in vitro compared with relatively poor in vivo activity, but found no evidence that the activity of the inhibitors could be improved by weakening the cell wall. Despite a number of drug discovery efforts attempting to develop inhibitors against PknB, it is yet to be reported that any such inhibitors prevent mycobacterial growth at submicromolar concentrations.
Embo Molecular Medicine | 2014
Conor McCloskey; Cara C. Rada; Elizabeth Bailey; Samantha McCavera; Hugo A. van den Berg; Jolene Atia; David A. Rand; Anatoly Shmygol; Yi-Wah Chan; Siobhan Quenby; Jan J. Brosens; Manu Vatish; Jie Zhang; Jerod S. Denton; Michael J. Taggart; Catherine A. Kettleborough; David Tickle; Jeff Jerman; Paul D. Wright; Timothy Dale; Srinivasan Kanumilli; Derek J. Trezise; Steve Thornton; Pamela Brown; Roberto Catalano; Nan Lin; Sarah K. England; Andrew M. Blanks
Abnormal uterine activity in pregnancy causes a range of important clinical disorders, including preterm birth, dysfunctional labour and post‐partum haemorrhage. Uterine contractile patterns are controlled by the generation of complex electrical signals at the myometrial smooth muscle plasma membrane. To identify novel targets to treat conditions associated with uterine dysfunction, we undertook a genome‐wide screen of potassium channels that are enriched in myometrial smooth muscle. Computational modelling identified Kir7.1 as potentially important in regulating uterine excitability during pregnancy. We demonstrate Kir7.1 current hyper‐polarizes uterine myocytes and promotes quiescence during gestation. Labour is associated with a decline, but not loss, of Kir7.1 expression. Knockdown of Kir7.1 by lentiviral expression of miRNA was sufficient to increase uterine contractile force and duration significantly. Conversely, overexpression of Kir7.1 inhibited uterine contractility. Finally, we demonstrate that the Kir7.1 inhibitor VU590 as well as novel derivative compounds induces profound, long‐lasting contractions in mouse and human myometrium; the activity of these inhibitors exceeds that of other uterotonic drugs. We conclude Kir7.1 regulates the transition from quiescence to contractions in the pregnant uterus and may be a target for therapies to control uterine contractility.
Expert Opinion on Drug Discovery | 2006
Michael Dalrymple; Debbie Taylor; Catherine A. Kettleborough; Justin S. Bryans; Roberto Solari
The movement of ideas and innovation from academia into the world of business has a long and fruitful history. Ironically, it might be argued that the recent pressure put on universities and basic research organisations to protect and exploit their intellectual property has, in many ways, created a less conducive environment to successful commercialisation than existed 30 years ago. This movement has been concurrent with the drift of the Pharmaceutical industry towards a more risk-averse R&D strategy in which it has increasingly concentrated its resources on a reductionist drug discovery process and later stage clinical development. In effect, these two strategies have created a discontinuity between academic scientific output and industry at a time when academia as a source of innovation is perhaps more important to industry than ever.
Biochemical and Biophysical Research Communications | 2013
Paul D. Wright; Gregory Weir; Jamie Cartland; David Tickle; Catherine A. Kettleborough; M. Zameel Cader; Jeff Jerman
TRESK is a two-pore domain potassium channel. Loss of function mutations have been linked to typical migraine with aura and due to TRESKs expression pattern and role in neuronal excitability it represents a promising therapeutic target. We developed a cell based assay using baculovirus transduced U20S cells to screen for activators of TRESK. Using a thallium flux system to measure TRESK channel activity we identified Cloxyquin as a novel activator. Cloxyquin was shown to have an EC50 of 3.8μM in the thallium assay and displayed good selectivity against other potassium channels tested. Activity was confirmed using whole cell patch electrophysiology, with Cloxyquin causing a near two fold increase in outward current. The strategy presented here will be used to screen larger compound libraries with the aim of identifying novel chemical series which may be developed into new migraine prophylactics.
Nature Communications | 2017
David A. Baker; Lindsay B. Stewart; Jonathan M. Large; Paul W. Bowyer; Keith H. Ansell; María Belén Jiménez-Díaz; Majida El Bakkouri; Kristian Birchall; Koen J. Dechering; Nathalie Bouloc; Peter J. Coombs; David Whalley; Denise J. Harding; Ela Smiljanic-Hurley; Mary Wheldon; Eloise M. Walker; Johannes T. Dessens; María J. Lafuente; Laura Sanz; Francisco-Javier Gamo; Santiago Ferrer; Raymond Hui; Teun Bousema; Iñigo Angulo-Barturen; Andy Merritt; Simon L. Croft; Winston Gutteridge; Catherine A. Kettleborough; Simon A. Osborne
To combat drug resistance, new chemical entities are urgently required for use in next generation anti-malarial combinations. We report here the results of a medicinal chemistry programme focused on an imidazopyridine series targeting the Plasmodium falciparum cyclic GMP-dependent protein kinase (PfPKG). The most potent compound (ML10) has an IC50 of 160 pM in a PfPKG kinase assay and inhibits P. falciparum blood stage proliferation in vitro with an EC50 of 2.1 nM. Oral dosing renders blood stage parasitaemia undetectable in vivo using a P. falciparum SCID mouse model. The series targets both merozoite egress and erythrocyte invasion, but crucially, also blocks transmission of mature P. falciparum gametocytes to Anopheles stephensi mosquitoes. A co-crystal structure of PvPKG bound to ML10, reveals intimate molecular contacts that explain the high levels of potency and selectivity we have measured. The properties of this series warrant consideration for further development to produce an antimalarial drug.Protein kinases are promising drug targets for treatment of malaria. Here, starting with a medicinal chemistry approach, Baker et al. generate an imidazopyridine that selectively targets Plasmodium falciparum PKG, inhibits blood stage parasite growth in vitro and in mice and blocks transmission to mosquitoes.
Journal of Biomolecular Screening | 2015
Paul D. Wright; Srinivasan Kanumilli; David Tickle; Jamie Cartland; Nathalie Bouloc; Timothy Dale; Derek J. Tresize; Conor McCloskey; Samantha McCavera; Andrew M. Blanks; Catherine A. Kettleborough; Jeffrey Jerman
Kir7.1 is an inwardly rectifying potassium channel that has been implicated in controlling the resting membrane potential of the myometrium. Abnormal uterine activity in pregnancy plays an important role in postpartum hemorrhage, and novel therapies for this condition may lie in manipulation of membrane potential. This work presents an assay development and screening strategy for identifying novel inhibitors of Kir7.1. A cell-based automated patch-clamp electrophysiology assay was developed using the IonWorks Quattro (Molecular Devices, Sunnyvale, CA) system, and the iterative optimization is described. In total, 7087 compounds were tested, with a hit rate (>40% inhibition) of 3.09%. During screening, average Z′ values of 0.63 ± 0.09 were observed. After chemistry triage, lead compounds were resynthesized and activity confirmed by IC50 determinations. The most potent compound identified (MRT00200769) gave rise to an IC50 of 1.3 µM at Kir7.1. Compounds were assessed for selectivity using the inwardly rectifying potassium channel Kir1.1 (ROMK) and hERG (human Ether-à-go-go Related Gene). Pharmacological characterization of known Kir7.1 inhibitors was also carried out and analogues of VU590 tested to assess selectivity at Kir7.1.
Biochemical and Biophysical Research Communications | 2017
Paul D. Wright; Emma L. Veale; David McCoull; David Tickle; Jonathan M. Large; Emma Ococks; Gemma Gothard; Catherine A. Kettleborough; Alistair Mathie; Jeffrey Jerman
Two-pore domain potassium channels (K2Ps) are characterized by their four transmembrane domain and two-pore topology. They carry background (or leak) potassium current in a variety of cell types. Despite a number of important roles there is currently a lack of pharmacological tools with which to further probe K2P function. We have developed a cell-based thallium flux assay, using baculovirus delivered TASK3 (TWIK-related acid-sensitive K+ channel 3, KCNK9, K2P9.1) with the aim of identifying novel, selective TASK3 activators. After screening a library of 1000 compounds, including drug-like and FDA approved molecules, we identified Terbinafine as an activator of TASK3. In a thallium flux assay a pEC50 of 6.2 ( ±0.12) was observed. When Terbinafine was screened against TASK2, TREK2, THIK1, TWIK1 and TRESK no activation was observed in thallium flux assays. Several analogues of Terbinafine were also purchased and structure activity relationships examined. To confirm Terbinafines activation of TASK3 whole cell patch clamp electrophysiology was carried out and clear potentiation observed in both the wild type channel and the pathophysiological, Birk-Barel syndrome associated, G236R TASK3 mutant. No activity at TASK1 was observed in electrophysiology studies. In conclusion, we have identified the first selective activator of the two-pore domain potassium channel TASK3.
Journal of Biomolecular Screening | 2013
Rachel White; Barbara Saxty; Jonathan M. Large; Catherine A. Kettleborough; Andrew P. Jackson
Ribonuclease H2 (RNase H2) is a nuclease that specifically hydrolyzes RNA residues in RNA-DNA hybrids. Mutations in the RNase H2 enzyme complex have been identified in the genetic disorder Aicardi-Goutières syndrome (AGS), which has similarities to the autoimmune disease systemic lupus eryrthrematosis (SLE). The RNase H2 enzyme has also been recently implicated as a key genome surveillance enzyme. Therefore, small-molecule modulators of RNase H2 activity may have utility in therapeutics and as tools to investigate the cellular functions of RNase H2. A fluorescent quench assay, measuring cleavage of an RNA-DNA duplex substrate by recombinant RNase H2, was developed into a high-throughput format and used to screen a 48 560 compound library. A hit validation strategy was subsequently employed, leading to the identification of two novel inhibitor compounds with in vitro nanomolar range inhibition of RNase H2 activity and >100-fold selectivity compared with RNase H type 1. These compounds are the first small-molecule inhibitors of RNase H2 to be reported. They and their derivatives should provide the basis for the development of tool compounds investigating the cellular functions of the RNase H2 enzyme, and, potentially, for pharmacological manipulation of nucleic acid–mediated immune responses.