Identification and Characterization of NDT 9513727 [N,N-bis(1,3-Benzodioxol-5-ylmethyl)-1-butyl-2,4-diphenyl-1H-imidazole-5-methanamine], a Novel, Orally Bioavailable C5a Receptor Inverse Agonist

Brodbeck, Robbin; Cortright, Daniel N.; Kieltyka, Andrzej; Yu, Jing; Baltazar, Carolyn; Buck, Matthew; Meade, Robin; Maynard, George D.; Thurkauf, Andrew; Chien, Du-Shieng; Hutchison, Alan J.; Krause, James E.

doi:10.1124/jpet.108.141572

Cited by 27 publications

(25 citation statements)

References 32 publications

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“…In the next step, we tested the potency of different commercially available C5aR1 antagonists to neutralize PVL and HlgCB toxicity. NDT 9513727 and W-54011 are small molecules with potent C5aR1 antagonistic properties (36, 37). PMX 205 and AS-65122 are C5amimetic peptides blocking the binding of C5a (38, 39).…”

Section: Resultsmentioning

confidence: 99%

Differential Interaction of the Staphylococcal Toxins Panton–Valentine Leukocidin and γ-Hemolysin CB with Human C5a Receptors

Spaan

Schiepers

Haas

et al. 2015

The Journal of Immunology

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Staphylococcus aureus is well adapted to the human host. Evasion of the host phagocyte response is critical for successful infection. The staphylococcal bicomponent pore-forming toxins Panton-Valentine Leukocidin (PVL) and γ-Hemolysin CB (HlgCB) target human phagocytes through interaction with the complement receptors C5aR1 and C5aR2. Currently, the apparent redundancy of both toxins cannot be adequately addressed in experimental models of infection since mice are resistant to PVL and HlgCB. The molecular basis for species specificity of the two toxins in animal models is not completely understood. We show that PVL and HlgCB feature distinct activity towards neutrophils of different mammalian species, where activity of PVL is found to be restricted to fewer species than that of HlgCB. Over-expression of various mammalian C5a receptors in HEK cells confirms that cytotoxicity towards neutrophils is driven by species-specific interactions of the toxins with C5aR1. By taking advantage of the species-specific engagement of the toxins with their receptors, we demonstrate that PVL and HlgCB differentially interact with human C5aR1 and C5aR2. In addition, binding studies illustrate that different parts of the receptor are involved in the initial binding of the toxin and the subsequent formation of lytic pores. These findings allow a better understanding of the molecular mechanism of pore formation. Finally, we show that toxicity of PVL, but not HlgCB, is neutralized by various C5aR1-antagonists. This study offers directions for the development of improved preclinical models for infection, and the design of drugs antagonizing leukocidin toxicity.

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Section: Resultsmentioning

confidence: 99%

Differential Interaction of the Staphylococcal Toxins Panton–Valentine Leukocidin and γ-Hemolysin CB with Human C5a Receptors

Spaan

Schiepers

Haas

et al. 2015

The Journal of Immunology

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“…We pretreated LAD2 cells with PTx, an inhibitor of Ga i proteins, and NDT 9513727, a C5aR1 antagonist, and measured C5a-desArg and C5a-desArg-induced mast cell adhesion to fibronectin. C5aR1 signals through Ga i and NDT 9513727 blocks C5aR1 signaling in other cell types (42). Pretreatment of LAD2 cells with either PTx (Fig.…”

Section: Inhibition Of C5ar1 Does Not Affect Complement-induced Mast mentioning

confidence: 99%

The Novel Receptor C5aR2 Is Required for C5a-Mediated Human Mast Cell Adhesion, Migration, and Proinflammatory Mediator Production

Pundir

MacDonald

Kulka

2015

The Journal of Immunology

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C5a generated during complement activation possesses proinflammatory and immunoregulatory properties critical for the development and modulation of allergic immune responses. In immune cells, C5a mediates its effects through binding to two G protein–coupled receptors, C5aR1 and C5aR2. Mast cells are key effectors in allergic reactions, and decades of research have suggested that the majority of C5a effects on mast cells are mediated through C5aR1, whereas the expression and function of C5aR2 have not been explored. We demonstrated that the human mast cell line Laboratory of Allergic Diseases 2 (LAD2) expresses surface C5aR2 but not C5aR1, whereas CD34+ cell–derived primary mast cells do not express surface C5aR1 or C5aR2. Stem cell factor and IL-4 upregulated C5aR2 expression on LAD2 cells. Furthermore, C5a caused internalization of LAD2 cell-surface C5aR2. We therefore used LAD2 cells as a model to study C5a/C5aR2-induced biological responses and signaling in human mast cells. We found that whereas C5a was unable to induce degranulation, it stimulated GM-CSF, TNF, CXCL10, and CCL2 production. C5a caused ERK phosphorylation, a signaling molecule important in cytokine and chemokine generation. In addition, C5a stimulated adhesion and chemotaxis of mast cells. Wortmannin, an inhibitor of PI3K, and small interfering RNA against β-arrestin-2 blocked C5a-induced adhesion. Silencing of C5aR2 using lentiviral short hairpin RNA rendered the cells unresponsive to C5a-induced adhesion, chemotaxis, and mediator release, as well as ERK phosphorylation. Overall, this study reveals a novel role for C5aR2 in C5a-mediated activation of mast cells and demonstrates that C5aR2 ligation initiates a β-arrestin-2–, PI3K-, and ERK-dependent signaling pathway in these cells.

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“…Because of inhibition of cytochrome P450 3A4, no further development of this drug occurred (Lee et al, 2008). 7C) is an inverse agonist at the human, primate, and gerbil C5a 1 receptor with little activity at the rodent C5a 1 receptor (Brodbeck et al, 2008). NDT 9513727 has a pIC 50 = 6.9 at that C5a 1 receptor but is not active at the C5a 2 receptor and is also orally bioavailable.…”

mentioning

confidence: 99%

International Union of Basic and Clinical Pharmacology. LXXXVII. Complement Peptide C5a, C4a, and C3a Receptors

Klos

Wende²,

Wareham³

et al. 2013

Pharmacol Rev

232

245

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Identification and Characterization of NDT 9513727 [N,N-bis(1,3-Benzodioxol-5-ylmethyl)-1-butyl-2,4-diphenyl-1H-imidazole-5-methanamine], a Novel, Orally Bioavailable C5a Receptor Inverse Agonist

Abstract: The complement system represents an innate immune mechanism of host defense that has three effector arms, the C3a receptor, the C5a receptor (C5aR), and the membrane attack complex.

Cited by 27 publications

References 32 publications

Differential Interaction of the Staphylococcal Toxins Panton–Valentine Leukocidin and γ-Hemolysin CB with Human C5a Receptors

Differential Interaction of the Staphylococcal Toxins Panton–Valentine Leukocidin and γ-Hemolysin CB with Human C5a Receptors

The Novel Receptor C5aR2 Is Required for C5a-Mediated Human Mast Cell Adhesion, Migration, and Proinflammatory Mediator Production

International Union of Basic and Clinical Pharmacology. LXXXVII. Complement Peptide C5a, C4a, and C3a Receptors

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