Walter E. DeWolf
GlaxoSmithKline
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Publication
Featured researches published by Walter E. DeWolf.
Journal of Biological Chemistry | 2000
Dennis Lee; Scott A. Long; Jerry L. Adams; George K. Chan; Kalindi Vaidya; Terry A. Francis; Kristine Kikly; James D. Winkler; Chiu-Mei Sung; Christine Debouck; Susan Richardson; Mark A. Levy; Walter E. DeWolf; Paul M. Keller; Thaddeus A. Tomaszek; Martha S. Head; M. Dominic Ryan; R. Curtis Haltiwanger; Po-Huang Liang; Cheryl A. Janson; Patrick McDevitt; Kyung Johanson; Nestor O. Concha; Winnie Chan; Sherin S. Abdel-Meguid; Alison M. Badger; Michael W. Lark; Daniel P. Nadeau; Larry J. Suva; Maxine Gowen
Caspases have been strongly implicated to play an essential role in apoptosis. A critical question regarding the role(s) of these proteases is whether selective inhibition of an effector caspase(s) will prevent cell death. We have identified potent and selective non-peptide inhibitors of the effector caspases 3 and 7. The inhibition of apoptosis and maintenance of cell functionality with a caspase 3/7-selective inhibitor is demonstrated for the first time, and suggests that targeting these two caspases alone is sufficient for blocking apoptosis. Furthermore, an x-ray co-crystal structure of the complex between recombinant human caspase 3 and an isatin sulfonamide inhibitor has been solved to 2.8-Å resolution. In contrast to previously reported peptide-based caspase inhibitors, the isatin sulfonamides derive their selectivity for caspases 3 and 7 by interacting primarily with the S2 subsite, and do not bind in the caspase primary aspartic acid binding pocket (S1). These inhibitors blocked apoptosis in murine bone marrow neutrophils and human chondrocytes. Furthermore, in camptothecin-induced chondrocyte apoptosis, cell functionality as measured by type II collagen promoter activity is maintained, an activity considered essential for cartilage homeostasis. These data suggest that inhibiting chondrocyte cell death with a caspase 3/7-selective inhibitor may provide a novel therapeutic approach for the prevention and treatment of osteoarthritis, or other disease states characterized by excessive apoptosis.
Antimicrobial Agents and Chemotherapy | 2002
David J. Payne; William H. Miller; Valerie Berry; John Brosky; Walter J. Burgess; Emile Chen; Walter E. DeWolf; Andrew Fosberry; Rebecca Greenwood; Martha S. Head; Dirk A. Heerding; Cheryl A. Janson; Deborah Dee Jaworski; Paul M. Keller; Peter J. Manley; Terrance D. Moore; Kenneth A. Newlander; Stewart Pearson; Brian J. Polizzi; Xiayang Qiu; Stephen Rittenhouse; Courtney Slater-Radosti; Kevin L. Salyers; Mark A. Seefeld; Martin G. Smyth; Dennis T. Takata; Irene Nijole Uzinskas; Kalindi Vaidya; Nicola G. Wallis; Scott B. Winram
ABSTRACT Bacterial enoyl-acyl carrier protein (ACP) reductase (FabI) catalyzes the final step in each elongation cycle of bacterial fatty acid biosynthesis and is an attractive target for the development of new antibacterial agents. High-throughput screening of the Staphylococcus aureus FabI enzyme identified a novel, weak inhibitor with no detectable antibacterial activity against S. aureus. Iterative medicinal chemistry and X-ray crystal structure-based design led to the identification of compound 4 [(E)-N-methyl-N-(2-methyl-1H-indol-3-ylmethyl)-3-(7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)acrylamide], which is 350-fold more potent than the original lead compound obtained by high-throughput screening in the FabI inhibition assay. Compound 4 has exquisite antistaphylococci activity, achieving MICs at which 90% of isolates are inhibited more than 500 times lower than those of nine currently available antibiotics against a panel of multidrug-resistant strains of S. aureus and Staphylococcus epidermidis. Furthermore, compound 4 exhibits excellent in vivo efficacy in an S. aureus infection model in rats. Biochemical and genetic approaches have confirmed that the mode of antibacterial action of compound 4 and related compounds is via inhibition of FabI. Compound 4 also exhibits weak FabK inhibitory activity, which may explain its antibacterial activity against Streptococcus pneumoniae and Enterococcus faecalis, which depend on FabK and both FabK and FabI, respectively, for their enoyl-ACP reductase function. These results show that compound 4 is representative of a new, totally synthetic series of antibacterial agents that has the potential to provide novel alternatives for the treatment of S. aureus infections that are resistant to our present armory of antibiotics.
The EMBO Journal | 2001
Ehsan Sharif‐Askari; Antoine Alam; Eric Rhéaume; Paul J. Beresford; Christian Scotto; Kamal Sharma; Dennis Lee; Walter E. DeWolf; Mark E. Nuttall; Judy Lieberman; Rafick-Pierre Sekaly
The protease granzyme B (GrB) plays a key role in the cytocidal activity during cytotoxic T lymphocyte (CTL)‐mediated programmed cell death. Multiple caspases have been identified as direct substrates for GrB, suggesting that the activation of caspases constitutes an important event during CTL‐induced cell death. However, recent studies have provided evidence for caspase‐independent pathway(s) during CTL‐mediated apoptosis. In this study, we demonstrate caspase‐independent and direct cleavage of the 45 kDa unit of DNA fragmentation factor (DFF45) by GrB both in vitro and in vivo. Using a novel and selective caspase‐3 inhibitor, we show the ability of GrB to process DFF45 directly and mediate DNA fragmentation in the absence of caspase‐3 activity. Furthermore, studies with DFF45 mutants reveal that both caspase‐3 and GrB share a common cleavage site, which is necessary and sufficient to induce DNA fragmentation in target cells during apoptosis. Together, our data suggest that CTLs possess alternative mechanism(s) for inducing DNA fragmentation without the requirement for caspases.
Bioorganic & Medicinal Chemistry Letters | 2001
Dirk A. Heerding; George M. Chan; Walter E. DeWolf; Andrew Fosberry; Cheryl A. Janson; Deborah D. Jaworski; Edward McManus; William Henry Miller; Terrance D. Moore; David J. Payne; Xiayang Qiu; Stephen Rittenhouse; Courtney Slater-Radosti; Ward W. Smith; Dennis T. Takata; Kalindi Vaidya; Catherine C.K. Yuan; William F. Huffman
1,4-Disubstituted imidazole inhibitors of Staphylococcus aureus and Escherichia coli enoyl acyl carrier protein reductase (FabI) have been identified. Crystal structure data shows the inhibitor 1 bound in the enzyme active site of E. coli FabI.
Biochemical Journal | 2003
Hedia Marrakchi; Walter E. DeWolf; Chad Quinn; Joshua West; Brian J. Polizzi; Chi Y. So; David J. Holmes; Shannon L. Reed; Richard J. Heath; David J. Payne; Charles O. Rock; Nicola G. Wallis
The enoyl-(acyl-carrier protein) (ACP) reductase catalyses the last step in each cycle of fatty acid elongation in the type II fatty acid synthase systems. An extensively characterized NADH-dependent reductase, FabI, is widely distributed in bacteria and plants, whereas the enoyl-ACP reductase, FabK, is a distinctly different member of this enzyme group discovered in Streptococcus pneumoniae. We were unable to delete the fabK gene from Strep. pneumoniae, suggesting that this is the only enoyl-ACP reductase in this organism. The FabK enzyme was purified and the biochemical properties of the reductase were examined. The visible absorption spectrum of the purified protein indicated the presence of a flavin cofactor that was identified as FMN by MS, and was present in a 1:1 molar ratio with protein. FabK specifically required NADH and the protein activity was stimulated by ammonium ions. FabK also exhibited NADH oxidase activity in the absence of substrate. Strep. pneumoniae belongs to the Bacillus / Lactobacillus / Streptococcus group that includes Staphylococcus aureus and Bacillus subtilis. These two organisms also contain FabK-related genes, suggesting that they may also express a FabK-like enoyl-ACP reductase. However, the genes did not complement a fabI (Ts) mutant and the purified flavoproteins were unable to reduce enoyl-ACP in vitro and did not exhibit NAD(P)H oxidase activity, indicating they were not enoyl-ACP reductases. The restricted occurrence of the FabK enoyl-ACP reductase may be related to the role of substrate-independent NADH oxidation in oxygen-dependent anaerobic energy metabolism.
Antimicrobial Agents and Chemotherapy | 2002
Frank Fan; Kang Yan; Nicola G. Wallis; Shannon L. Reed; Terrance D. Moore; Stephen Rittenhouse; Walter E. DeWolf; Jianzhong Huang; Damien McDevitt; William Henry Miller; Mark A. Seefeld; Kenneth A. Newlander; Dalia R. Jakas; Martha S. Head; David J. Payne
ABSTRACT The MICs of triclosan for 31 clinical isolates of Staphylococcus aureus were 0.016 μg/ml (24 strains), 1 to 2 μg/ml (6 strains), and 0.25 μg/ml (1 strain). All the strains for which triclosan MICs were elevated (>0.016 μg/ml) showed three- to fivefold increases in their levels of enoyl-acyl carrier protein (ACP) reductase (FabI) production. Furthermore, strains for which triclosan MICs were 1 to 2 μg/ml overexpressed FabI with an F204C alteration. Binding studies with radiolabeled NAD+ demonstrated that this change prevents the formation of the stable triclosan-NAD+-FabI complex, and both this alteration and its overexpression contributed to achieving MICs of 1 to 2 μg/ml for these strains. Three novel, potent inhibitors of FabI (50% inhibitory concentrations, ≤64 nM) demonstrated up to 1,000-fold better activity than triclosan against the strains for which triclosan MICs were elevated. None of the compounds tested from this series formed a stable complex with NAD+-FabI. Consequently, although the overexpression of wild-type FabI gave rise to an increase in the MICs, as expected, overexpression of FabI with an F204C alteration did not cause an additional increase in resistance. Therefore, this work identifies the mechanisms of triclosan resistance in S. aureus, and we present three compounds from a novel chemical series of FabI inhibitors which have excellent activities against both triclosan-resistant and -sensitive clinical isolates of S. aureus.
Bioorganic & Medicinal Chemistry Letters | 2001
Mark A. Seefeld; William Henry Miller; Kenneth A. Newlander; Walter J. Burgess; David J. Payne; Stephen Rittenhouse; Terrance D. Moore; Walter E. DeWolf; Paul M. Keller; Xiayang Qiu; Cheryl A. Janson; Kalindi Vaidya; Andrew Fosberry; Martin G. Smyth; Deborah D. Jaworski; Courtney Slater-Radosti; William F. Huffman
An SAR study of a screening lead has led to the identification of 2,9-disubstituted 1,2,3,4-tetrahydropyrido[3,4-b]indoles as inhibitors of Staphylococcus aureus enoyl acyl carrier protein reductase (FabI).
Bioorganic & Medicinal Chemistry Letters | 2008
Allen A. Thomas; J. De Meese; Y. Le Huerou; Steven Armen Boyd; Todd Romoff; Steven S. Gonzales; Indrani W. Gunawardana; Tomas Kaplan; Francis J. Sullivan; Kevin Ronald Condroski; Joseph P. Lyssikatos; Thomas Daniel Aicher; Josh Ballard; Bryan Bernat; Walter E. DeWolf; May Han; Christine Lemieux; Darin Smith; Solly Weiler; S. Kirk Wright; Guy Vigers; Barb Brandhuber
Inhibition of the thiamine-utilizing enzyme transketolase (TK) has been linked with diminished tumor cell proliferation. Most thiamine antagonists have a permanent positive charge on the B-ring, and it has been suggested that this charge is required for diphosphorylation by thiamine pyrophosphokinase (TPPK) and binding to TK. We sought to make neutral thiazolium replacements that would be substrates for TPPK, while not necessarily needing thiamine transporters (ThTr1 and ThTr2) for cell penetration. The synthesis, SAR, and structure-based rationale for highly potent non-thiazolium TK antagonists are presented.
Journal of Medicinal Chemistry | 2013
Ronald Jay Hinklin; Steven Armen Boyd; Mark Joseph Chicarelli; Kevin Ronald Condroski; Walter E. DeWolf; Patrice Lee; Wai-Man Lee; Ajay Singh; Laurie Thomas; Walter C. Voegtli; Lance Williams; Thomas Daniel Aicher
Glucose flux through glucokinase (GK) controls insulin release from the pancreas in response to high glucose concentrations. Glucose flux through GK also contributes to reducing hepatic glucose output. Because many individuals with type 2 diabetes appear to have an inadequacy or defect in one or both of these processes, compounds that can activate GK may serve as effective treatments for type 2 diabetes. Herein we report the identification and initial optimization of a novel series of allosteric glucokinase activators (GKAs). We discovered an initial thiazolylamino pyridine-based hit that was optimized using a structure-based design strategy and identified 26 as an early lead. Compound 26 demonstrated a good balance of in vitro potency and enzyme kinetic parameters and demonstrated blood glucose reductions in oral glucose tolerance tests in both C57BL/6J mice and high-fat fed Zucker diabetic fatty rats.
European Journal of Pharmacology | 2001
Victoria Y. Wong; Paul M. Keller; Mark E. Nuttall; Kristine Kikly; Walter E. DeWolf; Dennis Lee; Shujath M. Ali; Daniel P. Nadeau; Eugene T. Grygielko; Nicholas J. Laping; David P. Brooks
In the present study, we have used an in vitro model of apoptosis using primary human renal proximal tubular epithelial (RPTE) cells to investigate the mechanisms involved in renal cell apoptosis. Treatment of RPTE cells with okadaic acid for 24-48 h induced apoptosis in a concentration-dependent manner. Apoptosis was accompanied by the activation of the p38 mitogen-activated protein kinase (MAPK) pathway followed by the activation of caspase-9, -3, and -7. The induction of caspase activity correlated with the proteolytic cleavage of beta-catenin, suggesting that beta-catenin is a caspase substrate. The caspase inhibitor, Z-Val-Ala-Asp-fluoromethylketone (Z-VAD-fmk), resulted in a dose-dependent inhibition of apoptosis and beta-catenin cleavage. These data suggest that okadaic acid-induced apoptosis is p38 MAPK and caspase-dependent and that proteolytic cleavage of beta-catenin by caspases is likely to be a downstream molecular event associated with the morphological and cytoskeletal changes induced during apoptosis.