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Kathleen Deschamps, Wanda Cromlish,
Sean Weicker,
Sonia Lamontagne,
Sarah L Huszar,
Jacques Yves Gauthier,
John S Mudgett,
Alain Guimond,
Raymond Romand,
Nelly Frossard,
M David Percival,
Deborah Slipetz,
Christopher M Tan
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ABSTRACT: Cathepsin S (Cat S) is predominantly expressed in antigen-presenting cells and is up-regulated in several preclinical models of antigen-induced inflammation, suggesting a role in the allergic response. Prophylactic dosing of an irreversible Cat S inhibitor has been shown to attenuate pulmonary eosinophilia in mice, supporting the hypothesis that Cat S inhibition before the initiation of airway inflammation is beneficial in airway disease. In addition, Cat S has been shown to play a role in more distal events in the allergic response. To determine where Cat S inhibition may affect the allergic response, we used complementary genetic and pharmacological approaches to investigate the role of Cat S in the early and downstream allergic events in a murine model of antigen-induced lung inflammation. Cat S knockout mice did not develop ovalbumin-induced pulmonary inflammation, consistent with a role for Cat S in the development of the allergic response. Alternatively, wild-type mice were treated with a reversible, highly selective Cat S inhibitor in prophylactic and therapeutic dosing paradigms and assessed for changes in airway inflammation. Although both treatment paradigms resulted in potent Cat S inhibition, only prophylactic Cat S inhibitor dosing blocked lung inflammation, consistent with our findings in Cat S knockout mice. The findings indicate that although Cat S is up-regulated in allergic models, it does not appear to play a significant role in the downstream effector inflammatory phase in this model; however, our results demonstrate that Cat S inhibition in a prophylactic paradigm would ameliorate airway inflammation.
American Journal of Respiratory Cell and Molecular Biology 07/2011; 45(1):81-7. · 5.13 Impact Factor
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René St-Jacques,
Sylvie Toulmond,
Anick Auger,
Christoph Binkert, Wanda Cromlish,
Walter Fischli,
Jennifer Harris,
Patrick Hess,
Jie Lan,
Susana Liu,
Denis Riendeau,
Beat Steiner,
M David Percival
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ABSTRACT: The hypertensive double-transgenic (dTG) rat strain, expressing human renin and angiotensinogen, develops severe hypertension and organ damage and 50% of individuals die by 7 weeks of age. Here, we characterise a variation of this model in which animals present stable hypertension.
The effect of renin-angiotensin system blockers on blood pressure was determined with adult dTG rats treated with enalapril from 3 to 12 weeks of age. Tissue expression levels of renin and angiotensinogen were determined in dTG rats and rhesus monkeys by quantitative PCR.
Upon withdrawal from enalapril, mean arterial pressure (MAP) rose to 160-180 mmHg, with 95% of the female dTG rats surviving for 6 to 12 months, In Sprague-Dawley (SD) rats and rhesus monkeys, renin mRNA was absent or weakly expressed in most tissues, except for the kidneys and adrenals. In dTG rats, human renin expression was high in many additional tissues. The expression of human angiotensinogen in dTG rats followed a similar tissue pattern to SD and rhesus monkey angiotensinogen. Oral dosing of aliskiren, enalapril or losartan provided a similar maximal reduction in MAP and duration of efficacy in telemetrised dTG rats.
Enalapril-pretreated dTG rats are suitable for long-term MAP monitoring and sequential evaluation of human renin inhibitors.
Journal of Renin-Angiotensin-Aldosterone System 03/2011; 12(3):133-45. · 2.44 Impact Factor
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ABSTRACT: Renin is the first enzyme in the renin-angiotensin-aldosterone system which is the principal regulator of blood pressure and hydroelectrolyte balance. Previous studies suggest that cathepsin B is the activator of the prorenin zymogen. Here, we show no difference in plasma renin activity, or mean arterial blood pressure between wild-type and cathepsin B knockout mice. To account for potential gene compensation, a potent, selective, reversible cathepsin B inhibitor was developed to determine the role of cathepsin B on prorenin processing in rats. Pharmacological inhibition of cathepsin B in spontaneously hypertensive and double transgenic rats did not result in a reduction in renal mature renin protein levels or plasma renin activity. We conclude that cathepsin B does not play a significant role in this process in rodents.
Biological Chemistry 12/2010; 391(12):1469-73. · 2.96 Impact Factor
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Elise Isabel,
Kevin P Bateman,
Nathalie Chauret, Wanda Cromlish,
Sylvie Desmarais,
Le T Duong,
Jean-Pierre Falgueyret,
Jacques Yves Gauthier,
Sonia Lamontagne,
Cheuk K Lau, [......],
Joël Robichaud,
Gideon A Rodan,
Sevgi B Rodan,
Carmai Seto,
Michel Thérien,
Vouy Linh Truong,
Gregg Wesolowski,
Robert N Young,
Robert Zamboni,
W Cameron Black
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ABSTRACT: MK-0674 is a potent and selective cathepsin K inhibitor from the same structural class as odanacatib with a comparable inhibitory potency profile against Cat K. It is orally bioavailable and exhibits long half-life in pre-clinical species. In vivo studies using deuterated MK-0674 show stereoselective epimerization of the alcohol stereocenter via an oxidation/reduction cycle. From in vitro incubations, two metabolites could be identified: the hydroxyleucine and the glucuronide conjugate which were confirmed using authentic synthetic standards.
Bioorganic & medicinal chemistry letters 02/2010; 20(3):887-92. · 2.65 Impact Factor
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Jacques Yves Gauthier,
Nathalie Chauret, Wanda Cromlish,
Sylvie Desmarais,
Le T Duong,
Jean-Pierre Falgueyret,
Donald B Kimmel,
Sonia Lamontagne,
Serge Léger,
Tammy LeRiche, [......],
Gideon A Rodan,
Sevgi B Rodan,
Carmai Seto,
Michel Thérien,
Vouy-Linh Truong,
Michael C Venuti,
Gregg Wesolowski,
Robert N Young,
Robert Zamboni,
W Cameron Black
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ABSTRACT: Odanacatib is a potent, selective, and neutral cathepsin K inhibitor which was developed to address the metabolic liabilities of the Cat K inhibitor L-873724. Substituting P1 and modifying the P2 side chain led to a metabolically robust inhibitor with a long half-life in preclinical species. Odanacatib was more selective in whole cell assays than the published Cat K inhibitors balicatib and relacatib. Evaluation in dermal fibroblast culture showed minimal intracellular collagen accumulation relative to less selective Cat K inhibitors.
Bioorganic & medicinal chemistry letters 03/2008; 18(3):923-8. · 2.65 Impact Factor
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ABSTRACT: Although protein-tyrosine phosphatase 1B (PTP-1B) is a negative regulator of insulin action, adipose tissue from PTP-1B-/- mice does not show enhanced insulin-stimulated insulin receptor phosphorylation. Investigation of glucose uptake in isolated adipocytes revealed that the adipocytes from PTP-1B-/- mice have a significantly attenuated insulin response as compared with PTP-1B+/+ adipocytes. This insulin resistance manifests in PTP-1B-/- animals older than 16 weeks of age and could be partially rescued by adenoviral expression of PTP-1B in null adipocytes. Examination of adipose signaling pathways found that the basal p70S6K activity was at least 50% higher in adipose from PTP-1B-/- mice compared with wild type animals. The increased basal activity of p70S6K in PTP-1B-/- adipose correlated with decreases in IR substrate-1 protein levels and insulin-stimulated Akt/protein kinase B activity, explaining the decrease in insulin sensitivity even as insulin receptor phosphorylation was unaffected. The insulin resistance of the of the PTP-1B-/- adipocytes could also be rescued by treatment with rapamycin, suggesting that in adipose the loss of PTP-1B results in basal activation of mTOR (mammalian target of rapamycin) complex 1 leading to a tissue-specific insulin resistance.
Journal of Biological Chemistry 11/2007; 282(42):30423-33. · 4.77 Impact Factor
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Jacques Yves Gauthier,
W Cameron Black,
Isabelle Courchesne, Wanda Cromlish,
Sylvie Desmarais,
Robert Houle,
Sonia Lamontagne,
Chun Sing Li,
Frédéric Massé,
Daniel J McKay,
Marc Ouellet,
Joël Robichaud,
Jean-François Truchon,
Vouy-Linh Truong,
Qingping Wang,
M David Percival
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ABSTRACT: Highly potent, selective, and bioavailable inhibitors of human, mouse, or rat cathepsin S are described. The key structural features combine a sulfonyl moiety attached to a large group in P2 and a small substituent in P3.
Bioorganic & Medicinal Chemistry Letters 10/2007; 17(17):4929-33. · 2.55 Impact Factor
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Jean-Pierre Falgueyret,
Sylvie Desmarais,
Renata Oballa,
W Cameron Black, Wanda Cromlish,
Karine Khougaz,
Sonia Lamontagne,
Frederic Massé,
Denis Riendeau,
Sylvie Toulmond,
M David Percival
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ABSTRACT: The lysosomal cysteine protease cathepsin K is a target for osteoporosis therapy. The aryl-piperazine-containing cathepsin K inhibitor CRA-013783/L-006235 (1) displays greater than 4000-fold selectivity against the lysosomal/endosomal antitargets cathepsin B, L, and S. However, 1 and other aryl-piperazine-containing analogues, including balicatib (10), are approximately 10-100-fold more potent in cell-based enzyme occupancy assays than against each purified enzyme. This phenomenon arises from their basic, lipophilic nature, which results in lysosomal trapping. Consistent with its lysosomotropic nature, 1 accumulates in cells and in rat tissues of high lysosome content. In contrast, nonbasic aryl-morpholino-containing analogues do not exhibit lysosomotropic properties. Increased off-target activities of basic cathepsin K inhibitors were observed in a cell-based cathepsin S antigen presentation assay. No potency increases of basic inhibitors in a functional cathepsin K bone resorption whole cell assay were detected. Therefore, basic cathepsin K inhibitors, such as 1, suffer from reduced functional selectivities compared to those predicted using purified enzyme assays.
Journal of Medicinal Chemistry 01/2006; 48(24):7535-43. · 5.25 Impact Factor
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Jean-Pierre Falgueyret,
W Cameron Black, Wanda Cromlish,
Sylvie Desmarais,
Sonia Lamontagne,
Christophe Mellon,
Denis Riendeau,
Sevgi Rodan,
Paul Tawa,
Gregg Wesolowski,
Kathryn E Bass,
Shankar Venkatraman,
M David Percival
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ABSTRACT: We describe a novel diazomethylketone-containing irreversible inhibitor (BIL-DMK) which is specific for a subset of pharmaceutically important cysteine cathepsin proteases. BIL-DMK rapidly inactivates cathepsins B, F, K, L, S, and V in isolated enzyme assays and labels cathepsins in whole cells. The presence of catalytically active cathepsins B, L, and K or S was demonstrated using radioiodinated BIL-DMK in HepG2 (hepatoma), HIG82 (rabbit synoviocyte), and Ramos (B lymphoma) cell lines, respectively. The identity of each protein labeled was confirmed from the isoelectric point and molecular mass of the radioactive spots on two-dimensional gel and by comigration with each cathepsin as identified by immunoblotting. These cell lines were used to establish whole-cell enzyme occupancy assays to determine the potency of both irreversible and reversible inhibitors against each cathepsin in their native cellular lysosomal or endosomal environment. These whole-cell enzyme occupancy assays are useful to determine the cellular permeability of competing inhibitors and have the advantage of not requiring specific substrates for each cathepsin of interest.
Analytical Biochemistry 01/2005; 335(2):218-27. · 3.00 Impact Factor
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Cheuk K Lau,
Christopher I Bayly,
Jacques Yves Gauthier,
Chun Sing Li,
Michel Therien,
Ernest Asante-Appiah, Wanda Cromlish,
Yves Boie,
Farnaz Forghani,
Sylvie Desmarais,
Qingping Wang,
Kathryn Skorey,
Deena Waddleton,
Paul Payette,
Chidambaram Ramachandran,
Brian P Kennedy,
Giovana Scapin
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ABSTRACT: A series of benzotriazole phenyldifluoromethylphosphonic acids were found to be potent PTP-1B inhibitors. Molecular modeling on the X-ray crystal structure of the lead structure led to the design of potent PTP-1B inhibitors that show moderate selectivity against TC-PTP, a very closely related protein tyrosine phosphatase.
Bioorganic & Medicinal Chemistry Letters 03/2004; 14(4):1043-8. · 2.55 Impact Factor
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Claude Dufresne,
Patrick Roy,
Zhaoyin Wang,
Ernest Asante-Appiah, Wanda Cromlish,
Yves Boie,
Farnaz Forghani,
Sylvie Desmarais,
Qingping Wang,
Kathryn Skorey,
Deena Waddleton,
Chidambaram Ramachandran,
Brian P Kennedy,
Lijing Xu,
Robert Gordon,
Chi Chung Chan,
Yves Leblanc
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ABSTRACT: The SAR from our peptide libraries was exploited to design a series of potent deoxybenzoin PTP-1B inhibitors. The introduction of an ortho bromo substituent next to the difluoromethylphosphonate warhead gave up to 20-fold increase in potency compared to the desbromo analogues. In addition, these compounds were orally bioavailable and active in the animal models of non-insulin dependent diabetes mellitus (NIDDM).
Bioorganic & Medicinal Chemistry Letters 03/2004; 14(4):1039-42. · 2.55 Impact Factor
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Giovanna Scapin,
Sangita B Patel,
Joseph W Becker,
Qingping Wang,
Caroline Desponts,
Deena Waddleton,
Kathryn Skorey, Wanda Cromlish,
Christopher Bayly,
Michel Therien,
Jacques Yves Gauthier,
Chun Sing Li,
Cheuk K Lau,
Chidambaram Ramachandran,
Brian P Kennedy,
Ernest Asante-Appiah
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ABSTRACT: Protein tyrosine phosphatase 1B (PTP1B) has been implicated in the regulation of the insulin signaling pathway and represents an attractive target for the design of inhibitors in the treatment of type 2 diabetes and obesity. Inspection of the structure of PTP1B indicates that potent PTP1B inhibitors may be obtained by targeting a secondary aryl phosphate-binding site as well as the catalytic site. We report here the crystal structures of PTP1B in complex with first and second generation aryldifluoromethyl-phosphonic acid inhibitors. While all compounds bind in a previously unexploited binding pocket near the primary binding site, the second generation compounds also reach into the secondary binding site, and exhibit moderate selectivity for PTP1B over the closely related T-cell phosphatase. The molecular basis for the selectivity has been confirmed by single point mutation at position 52, where the two phosphatases differ by a phenylalanine-to-tyrosine switch. These compounds present a novel platform for the development of potent and selective PTP1B inhibitors.
Biochemistry 11/2003; 42(39):11451-9. · 3.42 Impact Factor
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W Cameron Black,
Christine Brideau,
Chi Chung Chan,
Stella Charleson, Wanda Cromlish,
Robert Gordon,
Erich L Grimm,
Gregory Hughes,
Serge Leger,
Chun Sing Li,
Denis Riendeau,
Michel Thérien,
Zhaoyin Wang,
Li Jing Xu,
Petpiboon Prasit
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ABSTRACT: The introduction of a hydroxyl group into the 5-position of the diaryl furanone system provides highly selective inhibitors of cyclooxygenase-2. These molecules can be converted into their sodium salts which are water soluble, facilitating intravenous formulation. These salts show excellent potency in rat models of pain, fever and inflammation.
Bioorganic & Medicinal Chemistry Letters 04/2003; 13(6):1195-8. · 2.55 Impact Factor
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Mounib Elchebly,
Paul Payette,
Eva Michaliszyn, Wanda Cromlish,
Susan Collins,
Ailsa Lee Loy,
Denis Normandin,
Alan Cheng,
Jean Himms-Hagen,
Chi-Chung Chan,
Chidambaram Ramachandran,
Michael J. Gresser,
Michel L. Tremblay,
Brian P. Kennedy
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ABSTRACT: Protein tyrosine phosphatase–1B (PTP-1B) has been implicated in the negative regulation of insulin signaling. Disruption of
the mouse homolog of the gene encoding PTP-1B yielded healthy mice that, in the fed state, had blood glucose concentrations
that were slightly lower and concentrations of circulating insulin that were one-half those of their PTP-1B+/+ littermates. The enhanced insulin sensitivity of the PTP-1B−/− mice was also evident in glucose and insulin tolerance tests. The PTP-1B−/− mice showed increased phosphorylation of the insulin receptor in liver and muscle tissue after insulin injection in comparison
to PTP-1B+/+ mice. On a high-fat diet, the PTP-1B−/− and PTP-1B+/− mice were resistant to weight gain and remained insulin sensitive, whereas the PTP-1B+/+ mice rapidly gained weight and became insulin resistant. These results demonstrate that PTP-1B has a major role in modulating
both insulin sensitivity and fuel metabolism, thereby establishing it as a potential therapeutic target in the treatment of
type 2 diabetes and obesity.
Science 03/1999; 283(5407):1544-1548. · 31.20 Impact Factor
