Davis, O.A.
Cheung, K.-.
Brennan, A.
Lloyd, M.G.
Rodrigues, M.J.
Pierrat, O.A.
Collie, G.W.
Le Bihan, Y.-.
Huckvale, R.
Harnden, A.C.
Varela, A.
Bright, M.D.
Eve, P.
Hayes, A.
Henley, A.T.
Carter, M.D.
McAndrew, P.C.
Talbot, R.
Burke, R.
van Montfort, R.L.
Raynaud, F.I.
Rossanese, O.W.
Meniconi, M.
Bellenie, B.R.
Hoelder, S.
(2022). Optimizing Shape Complementarity Enables the Discovery of Potent Tricyclic BCL6 Inhibitors. ,
Vol.65
(12),
pp. 8169-8190.
show 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..
Huckvale, R.
Harnden, A.C.
Cheung, K.-.
Pierrat, O.A.
Talbot, R.
Box, G.M.
Henley, A.T.
de Haven Brandon, A.K.
Hallsworth, A.E.
Bright, M.D.
Akpinar, H.A.
Miller, D.S.
Tarantino, D.
Gowan, S.
Hayes, A.
Gunnell, E.A.
Brennan, A.
Davis, O.A.
Johnson, L.D.
de Klerk, S.
McAndrew, C.
Le Bihan, Y.-.
Meniconi, M.
Burke, R.
Kirkin, V.
van Montfort, R.L.
Raynaud, F.I.
Rossanese, O.W.
Bellenie, B.R.
Hoelder, S.
(2022). Improved Binding Affinity and Pharmacokinetics Enable Sustained Degradation of BCL6 In Vivo. ,
Vol.65
(12),
pp. 8191-8207.
show 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..
Walters, Z.S.
Aladowicz, E.
Villarejo-Balcells, B.
Nugent, G.
Selfe, J.L.
Eve, P.
Blagg, J.
Rossanese, O.
Shipley, J.
(2021). Role for the Histone Demethylase KDM4B in Rhabdomyosarcoma via CDK6 and CCNA2: Compensation by KDM4A and Apoptotic Response of Targeting Both KDM4B and KDM4A. Cancers,
Vol.13
(7).
show abstract
Histone demethylases are epigenetic modulators that play key roles in regulating gene expression related to many critical cellular functions and are emerging as promising therapeutic targets in a number of tumor types. We previously identified histone demethylase family members as overexpressed in the pediatric sarcoma, rhabdomyosarcoma. Here we show high sensitivity of rhabdomyosarcoma cells to a pan-histone demethylase inhibitor, JIB-04 and identify a key role for the histone demethylase KDM4B in rhabdomyosarcoma cell growth through an RNAi-screening approach. Decreasing KDM4B levels affected cell cycle progression and transcription of G1/S and G2/M checkpoint genes including CDK6 and CCNA2 , which are bound by KDM4B in their promoter regions. However, after sustained knockdown of KDM4B, rhabdomyosarcoma cell growth recovered. We show that this can be attributed to acquired molecular compensation via recruitment of KDM4A to the promoter regions of CDK6 and CCNA2 that are otherwise bound by KDM4B. Furthermore, upfront silencing of both KDM4B and KDM4A led to RMS cell apoptosis, not seen by reducing either alone. To circumvent compensation and elicit stronger therapeutic responses, our study supports targeting histone demethylase sub-family proteins through selective poly-pharmacology as a therapeutic approach..
Bellenie, B.R.
Cheung, K.-.
Varela, A.
Pierrat, O.A.
Collie, G.W.
Box, G.M.
Bright, M.D.
Gowan, S.
Hayes, A.
Rodrigues, M.J.
Shetty, K.N.
Carter, M.
Davis, O.A.
Henley, A.T.
Innocenti, P.
Johnson, L.D.
Liu, M.
de Klerk, S.
Le Bihan, Y.-.
Lloyd, M.G.
McAndrew, P.C.
Shehu, E.
Talbot, R.
Woodward, H.L.
Burke, R.
Kirkin, V.
van Montfort, R.L.
Raynaud, F.I.
Rossanese, O.W.
Hoelder, S.
(2020). Achieving In Vivo Target Depletion through the Discovery and Optimization of Benzimidazolone BCL6 Degraders. Journal of medicinal chemistry,
Vol.63
(8),
pp. 4047-4068.
show 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..
Le Bihan, Y.-.
Lanigan, R.M.
Atrash, B.
McLaughlin, M.G.
Velupillai, S.
Malcolm, A.G.
England, K.S.
Ruda, G.F.
Mok, N.Y.
Tumber, A.
Tomlin, K.
Saville, H.
Shehu, E.
McAndrew, C.
Carmichael, L.
Bennett, J.M.
Jeganathan, F.
Eve, P.
Donovan, A.
Hayes, A.
Wood, F.
Raynaud, F.I.
Fedorov, O.
Brennan, P.E.
Burke, R.
van Montfort, R.L.
Rossanese, O.W.
Blagg, J.
Bavetsias, V.
(2019). C8-substituted pyrido[3,4-d]pyrimidin-4(3H)-ones: Studies towards the identification of potent, cell penetrant Jumonji C domain containing histone lysine demethylase 4 subfamily (KDM4) inhibitors, compound profiling in cell-based target engagement assays. European journal of medicinal chemistry,
Vol.177,
pp. 316-337.
show abstract
Residues in the histone substrate binding sites that differ between the KDM4 and KDM5 subfamilies were identified. Subsequently, a C8-substituted pyrido[3,4-d]pyrimidin-4(3H)-one series was designed to rationally exploit these residue differences between the histone substrate binding sites in order to improve affinity for the KDM4-subfamily over KDM5-subfamily enzymes. In particular, residues E169 and V313 (KDM4A numbering) were targeted. Additionally, conformational restriction of the flexible pyridopyrimidinone C8-substituent was investigated. These approaches yielded potent and cell-penetrant dual KDM4/5-subfamily inhibitors including 19a (KDM4A and KDM5B Ki = 0.004 and 0.007 μM, respectively). Compound cellular profiling in two orthogonal target engagement assays revealed a significant reduction from biochemical to cell-based activity across multiple analogues; this decrease was shown to be consistent with 2OG competition, and suggests that sub-nanomolar biochemical potency will be required with C8-substituted pyrido[3,4-d]pyrimidin-4(3H)-one compounds to achieve sub-micromolar target inhibition in cells..
Macdonald, J.D.
Chacón Simon, S.
Han, C.
Wang, F.
Shaw, J.G.
Howes, J.E.
Sai, J.
Yuh, J.P.
Camper, D.
Alicie, B.M.
Alvarado, J.
Nikhar, S.
Payne, W.
Aho, E.R.
Bauer, J.A.
Zhao, B.
Phan, J.
Thomas, L.R.
Rossanese, O.W.
Tansey, W.P.
Waterson, A.G.
Stauffer, S.R.
Fesik, S.W.
(2019). Discovery and Optimization of Salicylic Acid-Derived Sulfonamide Inhibitors of the WD Repeat-Containing Protein 5–MYC Protein–Protein Interaction. Journal of medicinal chemistry,
Vol.62
(24),
pp. 11232-11259.
Howes, J.E.
Akan, D.T.
Burns, M.C.
Rossanese, O.W.
Waterson, A.G.
Fesik, S.W.
(2018). Small Molecule–Mediated Activation of RAS Elicits Biphasic Modulation of Phospho-ERK Levels that Are Regulated through Negative Feedback on SOS1. Molecular cancer therapeutics,
Vol.17
(5),
pp. 1051-1060.
show abstract
Abstract
Oncogenic mutation of RAS results in aberrant cellular signaling and is responsible for more than 30% of all human tumors. Therefore, pharmacologic modulation of RAS has attracted great interest as a therapeutic strategy. Our laboratory has recently discovered small molecules that activate Son of Sevenless (SOS)–catalyzed nucleotide exchange on RAS and inhibit downstream signaling. Here, we describe how pharmacologically targeting SOS1 induced biphasic modulation of RAS-GTP and ERK phosphorylation levels, which we observed in a variety of cell lines expressing different RAS-mutant isoforms. We show that compound treatment caused an increase in phosphorylation at ERK consensus motifs on SOS1 that was not observed with the expression of a non-phosphorylatable S1178A SOS1 mutant or after pretreatment with an ERK inhibitor. Phosphorylation at S1178 on SOS1 is known to inhibit the association between SOS1 and GRB2 and disrupt SOS1 membrane localization. Consistent with this, we show that wild-type SOS1 and GRB2 dissociated in a time-dependent fashion in response to compound treatment, and conversely, this interaction was enhanced with the expression of an S1178A SOS1 mutant. Furthermore, in cells expressing either S1178A SOS1 or a constitutively membrane-bound CAAX box tagged SOS1 mutant, we observed elevated RAS-GTP levels over time in response to compound, as compared with the biphasic changes in RAS-GTP exhibited in cells expressing wild-type SOS1. These results suggest that small molecule targeting of SOS1 can elicit a biphasic modulation of RAS-GTP and phospho-ERK levels through negative feedback on SOS1 that regulates the interaction between SOS1 and GRB2. Mol Cancer Ther; 17(5); 1051–60. ©2018 AACR..
Burns, M.C.
Howes, J.E.
Sun, Q.
Little, A.J.
Camper, D.V.
Abbott, J.R.
Phan, J.
Lee, T.
Waterson, A.G.
Rossanese, O.W.
Fesik, S.W.
(2018). High-throughput screening identifies small molecules that bind to the RAS:SOS:RAS complex and perturb RAS signaling. Analytical biochemistry,
Vol.548,
pp. 44-52.
Abbott, J.R.
Patel, P.A.
Howes, J.E.
Akan, D.T.
Kennedy, J.P.
Burns, M.C.
Browning, C.F.
Sun, Q.
Rossanese, O.W.
Phan, J.
Waterson, A.G.
Fesik, S.W.
(2018). Discovery of Quinazolines That Activate SOS1-Mediated Nucleotide Exchange on RAS. Acs medicinal chemistry letters,
Vol.9
(9),
pp. 941-946.
show abstract
Proteins in the RAS family are important regulators of cellular signaling and, when mutated, can drive cancer pathogenesis. Despite considerable effort over the last 30 years, RAS proteins have proven to be recalcitrant therapeutic targets. One approach for modulating RAS signaling is to target proteins that interact with RAS, such as the guanine nucleotide exchange factor (GEF) son of sevenless homologue 1 (SOS1). Here, we report hit-to-lead studies on quinazoline-containing compounds that bind to SOS1 and activate nucleotide exchange on RAS. Using structure-based design, we refined the substituents attached to the quinazoline nucleus and built in additional interactions not present in the initial HTS hit. Optimized compounds activate nucleotide exchange at single-digit micromolar concentrations in vitro. In HeLa cells, these quinazolines increase the levels of RAS-GTP and cause signaling changes in the mitogen-activated protein kinase/extracellular regulated kinase (MAPK/ERK) pathway..
Hodges, T.R.
Abbott, J.R.
Little, A.J.
Sarkar, D.
Salovich, J.M.
Howes, J.E.
Akan, D.T.
Sai, J.
Arnold, A.L.
Browning, C.
Burns, M.C.
Sobolik, T.
Sun, Q.
Beesetty, Y.
Coker, J.A.
Scharn, D.
Stadtmueller, H.
Rossanese, O.W.
Phan, J.
Waterson, A.G.
McConnell, D.B.
Fesik, S.W.
(2018). Discovery and Structure-Based Optimization of Benzimidazole-Derived Activators of SOS1-Mediated Nucleotide Exchange on RAS. Journal of medicinal chemistry,
Vol.61
(19),
pp. 8875-8894.
Wang, F.
Jeon, K.O.
Salovich, J.M.
Macdonald, J.D.
Alvarado, J.
Gogliotti, R.D.
Phan, J.
Olejniczak, E.T.
Sun, Q.
Wang, S.
Camper, D.
Yuh, J.P.
Shaw, J.G.
Sai, J.
Rossanese, O.W.
Tansey, W.P.
Stauffer, S.R.
Fesik, S.W.
(2018). Discovery of Potent 2-Aryl-6,7-dihydro-5H-pyrrolo[1,2-a]imidazoles as WDR5-WIN-Site Inhibitors Using Fragment-Based Methods and Structure-Based Design. Journal of medicinal chemistry,
Vol.61
(13),
pp. 5623-5642.
Shaw, S.
Bian, Z.
Zhao, B.
Tarr, J.C.
Veerasamy, N.
Jeon, K.O.
Belmar, J.
Arnold, A.L.
Fogarty, S.A.
Perry, E.
Sensintaffar, J.L.
Camper, D.V.
Rossanese, O.W.
Lee, T.
Olejniczak, E.T.
Fesik, S.W.
(2018). Optimization of Potent and Selective Tricyclic Indole Diazepinone Myeloid Cell Leukemia-1 Inhibitors Using Structure-Based Design. Journal of medicinal chemistry,
Vol.61
(6),
pp. 2410-2421.
Abbott, J.R.
Hodges, T.R.
Daniels, R.N.
Patel, P.A.
Kennedy, J.P.
Howes, J.E.
Akan, D.T.
Burns, M.C.
Sai, J.
Sobolik, T.
Beesetty, Y.
Lee, T.
Rossanese, O.W.
Phan, J.
Waterson, A.G.
Fesik, S.W.
(2018). Discovery of Aminopiperidine Indoles That Activate the Guanine Nucleotide Exchange Factor SOS1 and Modulate RAS Signaling. Journal of medicinal chemistry,
Vol.61
(14),
pp. 6002-6017.
Hatch, S.B.
Yapp, C.
Montenegro, R.C.
Savitsky, P.
Gamble, V.
Tumber, A.
Ruda, G.F.
Bavetsias, V.
Fedorov, O.
Atrash, B.
Raynaud, F.
Lanigan, R.
Carmichael, L.
Tomlin, K.
Burke, R.
Westaway, S.M.
Brown, J.A.
Prinjha, R.K.
Martinez, E.D.
Oppermann, U.
Schofield, C.J.
Bountra, C.
Kawamura, A.
Blagg, J.
Brennan, P.E.
Rossanese, O.
Müller, S.
(2017). Assessing histone demethylase inhibitors in cells: lessons learned. Epigenetics & chromatin,
Vol.10,
pp. 9-?.
show abstract
Background Histone lysine demethylases (KDMs) are of interest as drug targets due to their regulatory roles in chromatin organization and their tight associations with diseases including cancer and mental disorders. The first KDM inhibitors for KDM1 have entered clinical trials, and efforts are ongoing to develop potent, selective and cell-active 'probe' molecules for this target class. Robust cellular assays to assess the specific engagement of KDM inhibitors in cells as well as their cellular selectivity are a prerequisite for the development of high-quality inhibitors. Here we describe the use of a high-content cellular immunofluorescence assay as a method for demonstrating target engagement in cells.Results A panel of assays for the Jumonji C subfamily of KDMs was developed to encompass all major branches of the JmjC phylogenetic tree. These assays compare compound activity against wild-type KDM proteins to a catalytically inactive version of the KDM, in which residues involved in the active-site iron coordination are mutated to inactivate the enzyme activity. These mutants are critical for assessing the specific effect of KDM inhibitors and for revealing indirect effects on histone methylation status. The reported assays make use of ectopically expressed demethylases, and we demonstrate their use to profile several recently identified classes of KDM inhibitors and their structurally matched inactive controls. The generated data correlate well with assay results assessing endogenous KDM inhibition and confirm the selectivity observed in biochemical assays with isolated enzymes. We find that both cellular permeability and competition with 2-oxoglutarate affect the translation of biochemical activity to cellular inhibition.Conclusions High-content-based immunofluorescence assays have been established for eight KDM members of the 2-oxoglutarate-dependent oxygenases covering all major branches of the JmjC-KDM phylogenetic tree. The usage of both full-length, wild-type and catalytically inactive mutant ectopically expressed protein, as well as structure-matched inactive control compounds, allowed for detection of nonspecific effects causing changes in histone methylation as a result of compound toxicity. The developed assays offer a histone lysine demethylase family-wide tool for assessing KDM inhibitors for cell activity and on-target efficacy. In addition, the presented data may inform further studies to assess the cell-based activity of histone lysine methylation inhibitors..
Lee, T.
Bian, Z.
Zhao, B.
Hogdal, L.J.
Sensintaffar, J.L.
Goodwin, C.M.
Belmar, J.
Shaw, S.
Tarr, J.C.
Veerasamy, N.
Matulis, S.M.
Koss, B.
Fischer, M.A.
Arnold, A.L.
Camper, D.V.
Browning, C.F.
Rossanese, O.W.
Budhraja, A.
Opferman, J.
Boise, L.H.
Savona, M.R.
Letai, A.
Olejniczak, E.T.
Fesik, S.W.
(2017). Discovery and biological characterization of potent myeloid cell leukemia‐1 inhibitors. Febs letters,
Vol.591
(1),
pp. 240-251.
Pelz, N.F.
Bian, Z.
Zhao, B.
Shaw, S.
Tarr, J.C.
Belmar, J.
Gregg, C.
Camper, D.V.
Goodwin, C.M.
Arnold, A.L.
Sensintaffar, J.L.
Friberg, A.
Rossanese, O.W.
Lee, T.
Olejniczak, E.T.
Fesik, S.W.
(2016). Discovery of 2-Indole-acylsulfonamide Myeloid Cell Leukemia 1 (Mcl-1) Inhibitors Using Fragment-Based Methods. Journal of medicinal chemistry,
Vol.59
(5),
pp. 2054-2066.
Kanu, N.
Cerone, M.A.
Goh, G.
Zalmas, L.-.
Bartkova, J.
Dietzen, M.
McGranahan, N.
Rogers, R.
Law, E.K.
Gromova, I.
Kschischo, M.
Walton, M.I.
Rossanese, O.W.
Bartek, J.
Harris, R.S.
Venkatesan, S.
Swanton, C.
(2016). DNA replication stress mediates APOBEC3 family mutagenesis in breast cancer. Genome biology,
Vol.17
(1),
pp. 185-?.
show abstract
Background The APOBEC3 family of cytidine deaminases mutate the cancer genome in a range of cancer types. Although many studies have documented the downstream effects of APOBEC3 activity through next-generation sequencing, less is known about their upstream regulation. In this study, we sought to identify a molecular basis for APOBEC3 expression and activation. Results HER2 amplification and PTEN loss promote DNA replication stress and APOBEC3B activity in vitro and correlate with APOBEC3 mutagenesis in vivo. HER2-enriched breast carcinomas display evidence of elevated levels of replication stress-associated DNA damage in vivo. Chemical and cytotoxic induction of replication stress, through aphidicolin, gemcitabine, camptothecin or hydroxyurea exposure, activates transcription of APOBEC3B via an ATR/Chk1-dependent pathway in vitro. APOBEC3B activation can be attenuated through repression of oncogenic signalling, small molecule inhibition of receptor tyrosine kinase signalling and alleviation of replication stress through nucleoside supplementation. Conclusion These data link oncogene, loss of tumour suppressor gene and drug-induced replication stress with APOBEC3B activity, providing new insights into how cytidine deaminase-induced mutagenesis might be activated in tumourigenesis and limited therapeutically..
Patrone, J.D.
Pelz, N.F.
Bates, B.S.
Souza-Fagundes, E.M.
Vangamudi, B.
Camper, D.V.
Kuznetsov, A.G.
Browning, C.F.
Feldkamp, M.D.
Frank, A.O.
Gilston, B.A.
Olejniczak, E.T.
Rossanese, O.W.
Waterson, A.G.
Chazin, W.J.
Fesik, S.W.
(2016). Identification and Optimization of Anthranilic Acid Based Inhibitors of Replication Protein A. Chemmedchem,
Vol.11
(8),
pp. 893-899.
Goodwin, C.M.
Rossanese, O.W.
Olejniczak, E.T.
Fesik, S.W.
(2015). Myeloid cell leukemia-1 is an important apoptotic survival factor in triple-negative breast cancer. Cell death & differentiation,
Vol.22
(12),
pp. 2098-2106.
Waterson, A.G.
Kennedy, J.P.
Patrone, J.D.
Pelz, N.F.
Feldkamp, M.D.
Frank, A.O.
Vangamudi, B.
Souza-Fagundes, E.M.
Rossanese, O.W.
Chazin, W.J.
Fesik, S.W.
(2015). Diphenylpyrazoles as Replication Protein A Inhibitors. Acs medicinal chemistry letters,
Vol.6
(2),
pp. 140-145.
Burke, J.P.
Bian, Z.
Shaw, S.
Zhao, B.
Goodwin, C.M.
Belmar, J.
Browning, C.F.
Vigil, D.
Friberg, A.
Camper, D.V.
Rossanese, O.W.
Lee, T.
Olejniczak, E.T.
Fesik, S.W.
(2015). Discovery of Tricyclic Indoles That Potently Inhibit Mcl-1 Using Fragment-Based Methods and Structure-Based Design. Journal of medicinal chemistry,
Vol.58
(9),
pp. 3794-3805.
Frank, A.O.
Vangamudi, B.
Feldkamp, M.D.
Souza-Fagundes, E.M.
Luzwick, J.W.
Cortez, D.
Olejniczak, E.T.
Waterson, A.G.
Rossanese, O.W.
Chazin, W.J.
Fesik, S.W.
(2014). Discovery of a potent stapled helix peptide that binds to the 70N domain of replication protein A. Journal of medicinal chemistry,
Vol.57
(6),
pp. 2455-2461.
show abstract
Stapled helix peptides can serve as useful tools for inhibiting protein-protein interactions but can be difficult to optimize for affinity. Here we describe the discovery and optimization of a stapled helix peptide that binds to the N-terminal domain of the 70 kDa subunit of replication protein A (RPA70N). In addition to applying traditional optimization strategies, we employed a novel approach for efficiently designing peptides containing unnatural amino acids. We discovered hot spots in the target protein using a fragment-based screen, identified the amino acid that binds to the hot spot, and selected an unnatural amino acid to incorporate, based on the structure-activity relationships of small molecules that bind to this site. The resulting stapled helix peptide potently and selectively binds to RPA70N, does not disrupt ssDNA binding, and penetrates cells. This peptide may serve as a probe to explore the therapeutic potential of RPA70N inhibition in cancer..
Burns, M.C.
Sun, Q.
Daniels, R.N.
Camper, D.
Kennedy, J.P.
Phan, J.
Olejniczak, E.T.
Lee, T.
Waterson, A.G.
Rossanese, O.W.
Fesik, S.W.
(2014). Approach for targeting Ras with small molecules that activate SOS-mediated nucleotide exchange. Proceedings of the national academy of sciences,
Vol.111
(9),
pp. 3401-3406.
show abstract
Significance
Ras is one of the most highly validated targets in cancer; however, the discovery of potent inhibitors of Ras has been difficult to achieve. We report the discovery of small molecules that bind to a pocket on the Ras:Son of Sevenless:Ras complex and alter Ras activity in biochemical and cell-based experiments. High-resolution cocrystal structures define the protein–ligand interactions, and the lead compounds provide a starting point for the discovery of potent inhibitors of Ras signaling..
Patrone, J.D.
Kennedy, J.P.
Frank, A.O.
Feldkamp, M.D.
Vangamudi, B.
Pelz, N.F.
Rossanese, O.W.
Waterson, A.G.
Chazin, W.J.
Fesik, S.W.
(2013). Discovery of Protein–Protein Interaction Inhibitors of Replication Protein A. Acs medicinal chemistry letters,
Vol.4
(7),
pp. 601-605.
Rheault, T.R.
Stellwagen, J.C.
Adjabeng, G.M.
Hornberger, K.R.
Petrov, K.G.
Waterson, A.G.
Dickerson, S.H.
Mook, R.A.
Laquerre, S.G.
King, A.J.
Rossanese, O.W.
Arnone, M.R.
Smitheman, K.N.
Kane-Carson, L.S.
Han, C.
Moorthy, G.S.
Moss, K.G.
Uehling, D.E.
(2013). Discovery of Dabrafenib: A Selective Inhibitor of Raf Kinases with Antitumor Activity against B-Raf-Driven Tumors. Acs medicinal chemistry letters,
Vol.4
(3),
pp. 358-362.
Frank, A.O.
Feldkamp, M.D.
Kennedy, J.P.
Waterson, A.G.
Pelz, N.F.
Patrone, J.D.
Vangamudi, B.
Camper, D.V.
Rossanese, O.W.
Chazin, W.J.
Fesik, S.W.
(2013). Discovery of a Potent Inhibitor of Replication Protein A Protein–Protein Interactions Using a Fragment-Linking Approach. Journal of medicinal chemistry,
Vol.56
(22),
pp. 9242-9250.
Souza-Fagundes, E.M.
Frank, A.O.
Feldkamp, M.D.
Dorset, D.C.
Chazin, W.J.
Rossanese, O.W.
Olejniczak, E.T.
Fesik, S.W.
(2012). A high-throughput fluorescence polarization anisotropy assay for the 70N domain of replication protein A. Analytical biochemistry,
Vol.421
(2),
pp. 742-749.
Sun, Q.
Burke, J.P.
Phan, J.
Burns, M.C.
Olejniczak, E.T.
Waterson, A.G.
Lee, T.
Rossanese, O.W.
Fesik, S.W.
(2012). Discovery of Small Molecules that Bind to K-Ras and Inhibit Sos-Mediated Activation. Angewandte chemie international edition,
Vol.51
(25),
pp. 6140-6143.
Stellwagen, J.C.
Adjabeng, G.M.
Arnone, M.R.
Dickerson, S.H.
Han, C.
Hornberger, K.R.
King, A.J.
Mook, R.A.
Petrov, K.G.
Rheault, T.R.
Rominger, C.M.
Rossanese, O.W.
Smitheman, K.N.
Waterson, A.G.
Uehling, D.E.
(2011). Development of potent B-RafV600E inhibitors containing an arylsulfonamide headgroup. Bioorganic & medicinal chemistry letters,
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Losev, E.
Papanikou, E.
Rossanese, O.W.
Glick, B.S.
(2008). Cdc1p Is an Endoplasmic Reticulum-Localized Putative Lipid Phosphatase That Affects Golgi Inheritance and Actin Polarization by Activating Ca
2+
Signaling. Molecular and cellular biology,
Vol.28
(10),
pp. 3336-3343.
show abstract
ABSTRACT
In the budding yeast
Saccharomyces cerevisiae
, mutations in the essential gene
CDC1
cause defects in Golgi inheritance and actin polarization. However, the biochemical function of Cdc1p is unknown. Previous work showed that
cdc1
mutants accumulate intracellular Ca
2+
and display enhanced sensitivity to the extracellular Mn
2+
concentration, suggesting that Cdc1p might regulate divalent cation homeostasis. By contrast, our data indicate that Cdc1p is a Mn
2+
-dependent protein that can affect Ca
2+
levels. We identified a
cdc1
allele that activates Ca
2+
signaling but does not show enhanced sensitivity to the Mn
2+
concentration. Furthermore, our studies show that Cdc1p is an endoplasmic reticulum-localized transmembrane protein with a putative phosphoesterase domain facing the lumen.
cdc1
mutant cells accumulate an unidentified phospholipid, suggesting that Cdc1p may be a lipid phosphatase. Previous work showed that deletion of the plasma membrane Ca
2+
channel Cch1p partially suppressed the
cdc1
growth phenotype, and we find that deletion of Cch1p also suppresses the Golgi inheritance and actin polarization phenotypes. The combined data fit a model in which the
cdc1
mutant phenotypes result from accumulation of a phosphorylated lipid that activates Ca
2+
signaling.
.
Patel, J.C.
Rossanese, O.W.
Galán, J.E.
(2005). The functional interface between Salmonella and its host cell: opportunities for therapeutic intervention. Trends in pharmacological sciences,
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Rossanese, O.W.
Glick, B.S.
(2001). Deconstructing Golgi Inheritance. Traffic,
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pp. 589-596.
ROSSANESE, O.W.
(2001). A role for actin, Cdc1p, and Myo2p in the inheritance of late Golgi elements in Saccharomyces cerevisiae. J. cell biol.,
Vol.153,
pp. 47-62.
ROSSANESE, O.W.
(1999). Golgi structure correlated wtih transitional endoplasmic reticulum organization in Pichia pastoris and Saccharomyces cerevisiae. J. cell biol.,
Vol.145,
pp. 69-81.
Séron, K.
Tieaho, V.
Prescianotto-Baschong, C.
Aust, T.
Blondel, M.-.
Guillaud, P.
Devilliers, G.
Rossanese, O.W.
Glick, B.S.
Riezman, H.
Keränen, S.
Haguenauer-Tsapis, R.
(1998). A Yeast t-SNARE Involved in Endocytosis. Molecular biology of the cell,
Vol.9
(10),
pp. 2873-2889.
show abstract
The ORF YOL018c (TLG2) of Saccharomyces cerevisiae encodes a protein that belongs to the syntaxin protein family. The proteins of this family, t-SNAREs, are present on target organelles and are thought to participate in the specific interaction between vesicles and acceptor membranes in intracellular membrane trafficking. TLG2 is not an essential gene, and its deletion does not cause defects in the secretory pathway. However, its deletion in cells lacking the vacuolar ATPase subunit Vma2p leads to loss of viability, suggesting that Tlg2p is involved in endocytosis. In tlg2Δ cells, internalization was normal for two endocytic markers, the pheromone α-factor and the plasma membrane uracil permease. In contrast, degradation of α-factor and uracil permease was delayed intlg2Δ cells. Internalization of positively charged Nanogold shows that the endocytic pathway is perturbed in the mutant, which accumulates Nanogold in primary endocytic vesicles and shows a greatly reduced complement of early endosomes. These results strongly suggest that Tlg2p is a t-SNARE involved in early endosome biogenesis..
Sears, I.B.
O'Connor, J.
Rossanese, O.W.
Glick, B.S.
(1998). A versatile set of vectors for constitutive and regulated gene expression inPichia pastoris. Yeast,
Vol.14
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pp. 783-790.
Jiang, X.
Rossanese, O.W.
Brown, N.F.
Kujat-Choy, S.
Galán, J.E.
Finlay, B.B.
Brumell, J.H.
The related effector proteins SopD and SopD2 from Salmonella enterica serovar Typhimurium contribute to virulence during systemic infection of mice. Molecular microbiology,
Vol.54
(5),
pp. 1186-1198.
Bhave, M.
Papanikou, E.
Iyer, P.
Pandya, K.
Jain, B.K.
Ganguly, A.
Sharma, C.
Pawar, K.
Austin, J.
Day, K.J.
Rossanese, O.W.
Glick, B.S.
Bhattacharyya, D.
Golgi enlargement in Arf-depleted yeast cells is due to altered dynamics of cisternal maturation. Journal of cell science,
.
show abstract
Regulation of the size and abundance of membrane compartments is a fundamental cellular activity. In Saccharomyces cerevisiae, disruption of the ARF1 gene yields larger and fewer Golgi cisternae by partially depleting the Arf GTPase. We observe a similar phenotype with a thermosensitive mutation in Nmt1, which myristoylates and activates Arf. Therefore, partial depletion of Arf is a convenient tool for dissecting mechanisms that regulate Golgi structure. We find that in arf1Δ cells, late Golgi structure is particularly abnormal, with the number of late Golgi cisternae being severely reduced. This effect can be explained by selective changes in cisternal maturation kinetics. The arf1Δ mutation causes early Golgi cisternae to mature more slowly and less frequently, but does not alter the maturation of late Golgi cisternae. These changes quantitatively explain why late Golgi cisternae are fewer in number and correspondingly larger. With a stacked Golgi, similar changes in maturation kinetics could be used by the cell to modulate the number of cisternae per stack. Thus, the rates of processes that transform a maturing compartment can determine compartmental size and copy number..
Lloyd, M.G.
Huckvale, R.
Cheung, K.-.
Rodrigues, M.J.
Collie, G.W.
Pierrat, O.A.
Gatti Iou, M.
Carter, M.
Davis, O.A.
McAndrew, P.C.
Gunnell, E.
Le Bihan, Y.-.
Talbot, R.
Henley, A.T.
Johnson, L.D.
Hayes, A.
Bright, M.D.
Raynaud, F.I.
Meniconi, M.
Burke, R.
van Montfort, R.L.
Rossanese, O.W.
Bellenie, B.R.
Hoelder, S.
Into Deep Water: Optimizing BCL6 Inhibitors by Growing into a Solvated Pocket. Journal of medicinal chemistry,
Vol.64
(23),
pp. 17079-17097.
show abstract
We describe the optimization of modestly active starting points to potent inhibitors of BCL6 by growing into a subpocket, which was occupied by a network of five stably bound water molecules. Identifying potent inhibitors required not only forming new interactions in the subpocket but also perturbing the water network in a productive, potency-increasing fashion while controlling the physicochemical properties. We achieved this goal in a sequential manner by systematically probing the pocket and the water network, ultimately achieving a 100-fold improvement of activity. The most potent compounds displaced three of the five initial water molecules and formed hydrogen bonds with the remaining two. Compound 25 showed a promising profile for a lead compound with submicromolar inhibition of BCL6 in cells and satisfactory pharmacokinetic (PK) properties. Our work highlights the importance of finding productive ways to perturb existing water networks when growing into solvent-filled protein pockets..
Miller, D.
Voell, S.A.
Sosič, I.
Proj, M.
Rossanese, O.
Schnakenburg, G.
Gütschow, M.
Collins, I.
Steinebach, C.
Encoding BRAF Inhibitor Functions in Protein Degraders. Rsc medicinal chemistry,
.
show abstract
Various BRAF kinase inhibitors were developed to treat cancers carrying the BRAFV600E mutation. First-generation BRAF inhibitors could lead to paradoxical activation of the MAPK pathway, limiting their clinical usefulness. Here,....