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Receptor residence time trumps drug-likeness and oral bioavailability in determining efficacy of complement C5a antagonists.

Seow V, Lim J, Cotterell AJ, Yau MK, Xu W, Lohman RJ, Kok WM, Stoermer MJ, Sweet MJ, Reid RC, Suen JY, Fairlie DP - Sci Rep (2016)

Bottom Line: The unusually long residence time of 3D53 on its receptor was mechanistically probed by molecular dynamics simulations, which revealed long-lasting interactions that trap the antagonist within the receptor.Despite negligible oral bioavailability, 3D53 was much more orally efficacious than W54011 or JJ47 in preventing repeated agonist insults to induce rat paw oedema over 24 h.Thus, residence time on a receptor can trump drug-likeness in determining efficacy, even oral efficacy, of pharmacological agents.

View Article: PubMed Central - PubMed

Affiliation: Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia.

ABSTRACT
Drug discovery and translation are normally based on optimizing efficacy by increasing receptor affinity, functional potency, drug-likeness (rule-of-five compliance) and oral bioavailability. Here we demonstrate that residence time of a compound on its receptor has an overriding influence on efficacy, exemplified for antagonists of inflammatory protein complement C5a that activates immune cells and promotes disease. Three equipotent antagonists (3D53, W54011, JJ47) of inflammatory responses to C5a (3 nM) were compared for drug-likeness, receptor affinity and antagonist potency in human macrophages, and anti-inflammatory efficacy in rats. Only the least drug-like antagonist (3D53) maintained potency in cells against higher C5a concentrations and had a much longer duration of action (t1/2 ~ 20 h) than W54011 or JJ47 (t1/2 ~ 1 -3 h) in inhibiting macrophage responses. The unusually long residence time of 3D53 on its receptor was mechanistically probed by molecular dynamics simulations, which revealed long-lasting interactions that trap the antagonist within the receptor. Despite negligible oral bioavailability, 3D53 was much more orally efficacious than W54011 or JJ47 in preventing repeated agonist insults to induce rat paw oedema over 24 h. Thus, residence time on a receptor can trump drug-likeness in determining efficacy, even oral efficacy, of pharmacological agents.

No MeSH data available.


Related in: MedlinePlus

Antagonism of C5a-induced gene expression in human and rat macrophages by 3D53, W54011 and JJ47.(A,B) HMDM were stimulated with 10 nM C5a and lysed after 30 min. For antagonism of C5aR, HMDM were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to 10 nM C5a stimulation after (A) 1 h or (B) 16 h post-treatment with antagonist. (C) Rat macrophages were stimulated with 0.1 μM C5a and cells are lysed after 30 min. For antagonism of C5aR, cells were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to C5a treatment after 1 h post-treatment with antagonist. Gene expression was measured by real-time PCR, normalized against 18S and converted to fold change relative to control. Error bars are means ± SEM of three independent experiments (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.
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f5: Antagonism of C5a-induced gene expression in human and rat macrophages by 3D53, W54011 and JJ47.(A,B) HMDM were stimulated with 10 nM C5a and lysed after 30 min. For antagonism of C5aR, HMDM were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to 10 nM C5a stimulation after (A) 1 h or (B) 16 h post-treatment with antagonist. (C) Rat macrophages were stimulated with 0.1 μM C5a and cells are lysed after 30 min. For antagonism of C5aR, cells were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to C5a treatment after 1 h post-treatment with antagonist. Gene expression was measured by real-time PCR, normalized against 18S and converted to fold change relative to control. Error bars are means ± SEM of three independent experiments (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.

Mentions: Real-time PCR analyses were conducted to further check the effect of residence time in other assays. All three antagonists were found to block C5a-induced expression of the inflammatory genes TNF, IL1B, CCL3 and PTGS2 in HMDM at 1 h post-antagonism, while W54011 and JJ47 blocked C5a-induced CCL3 gene expression to a lesser extent (Fig. 5A). At 16 h post-antagonist treatment, 3D53 still blocked or significantly inhibited C5a-mediated expression of these human genes, whereas W54011 and JJ47 did not (Fig. 5B). Together, these findings are consistent with W54011 and JJ47 having much shorter durations of action due to much shorter residence times on C5aR of HMDM, being completely displaced from the receptor well within 16 h. Although the responses from human C5a on rat macrophages were not as potent across the same panel of genes, all three antagonists were able to block C5a-mediated responses at 1 h post-antagonism (Fig. 5C). This suggested the feasibility of comparing in vivo antagonist activity of 3D53, W54011 and JJ47 in an animal model of inflammation.


Receptor residence time trumps drug-likeness and oral bioavailability in determining efficacy of complement C5a antagonists.

Seow V, Lim J, Cotterell AJ, Yau MK, Xu W, Lohman RJ, Kok WM, Stoermer MJ, Sweet MJ, Reid RC, Suen JY, Fairlie DP - Sci Rep (2016)

Antagonism of C5a-induced gene expression in human and rat macrophages by 3D53, W54011 and JJ47.(A,B) HMDM were stimulated with 10 nM C5a and lysed after 30 min. For antagonism of C5aR, HMDM were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to 10 nM C5a stimulation after (A) 1 h or (B) 16 h post-treatment with antagonist. (C) Rat macrophages were stimulated with 0.1 μM C5a and cells are lysed after 30 min. For antagonism of C5aR, cells were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to C5a treatment after 1 h post-treatment with antagonist. Gene expression was measured by real-time PCR, normalized against 18S and converted to fold change relative to control. Error bars are means ± SEM of three independent experiments (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f5: Antagonism of C5a-induced gene expression in human and rat macrophages by 3D53, W54011 and JJ47.(A,B) HMDM were stimulated with 10 nM C5a and lysed after 30 min. For antagonism of C5aR, HMDM were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to 10 nM C5a stimulation after (A) 1 h or (B) 16 h post-treatment with antagonist. (C) Rat macrophages were stimulated with 0.1 μM C5a and cells are lysed after 30 min. For antagonism of C5aR, cells were pretreated with 1 μM 3D53, W54011 or JJ47 for 1 h. Excess unbound antagonists were removed by washing and subjected to C5a treatment after 1 h post-treatment with antagonist. Gene expression was measured by real-time PCR, normalized against 18S and converted to fold change relative to control. Error bars are means ± SEM of three independent experiments (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.
Mentions: Real-time PCR analyses were conducted to further check the effect of residence time in other assays. All three antagonists were found to block C5a-induced expression of the inflammatory genes TNF, IL1B, CCL3 and PTGS2 in HMDM at 1 h post-antagonism, while W54011 and JJ47 blocked C5a-induced CCL3 gene expression to a lesser extent (Fig. 5A). At 16 h post-antagonist treatment, 3D53 still blocked or significantly inhibited C5a-mediated expression of these human genes, whereas W54011 and JJ47 did not (Fig. 5B). Together, these findings are consistent with W54011 and JJ47 having much shorter durations of action due to much shorter residence times on C5aR of HMDM, being completely displaced from the receptor well within 16 h. Although the responses from human C5a on rat macrophages were not as potent across the same panel of genes, all three antagonists were able to block C5a-mediated responses at 1 h post-antagonism (Fig. 5C). This suggested the feasibility of comparing in vivo antagonist activity of 3D53, W54011 and JJ47 in an animal model of inflammation.

Bottom Line: The unusually long residence time of 3D53 on its receptor was mechanistically probed by molecular dynamics simulations, which revealed long-lasting interactions that trap the antagonist within the receptor.Despite negligible oral bioavailability, 3D53 was much more orally efficacious than W54011 or JJ47 in preventing repeated agonist insults to induce rat paw oedema over 24 h.Thus, residence time on a receptor can trump drug-likeness in determining efficacy, even oral efficacy, of pharmacological agents.

View Article: PubMed Central - PubMed

Affiliation: Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia.

ABSTRACT
Drug discovery and translation are normally based on optimizing efficacy by increasing receptor affinity, functional potency, drug-likeness (rule-of-five compliance) and oral bioavailability. Here we demonstrate that residence time of a compound on its receptor has an overriding influence on efficacy, exemplified for antagonists of inflammatory protein complement C5a that activates immune cells and promotes disease. Three equipotent antagonists (3D53, W54011, JJ47) of inflammatory responses to C5a (3 nM) were compared for drug-likeness, receptor affinity and antagonist potency in human macrophages, and anti-inflammatory efficacy in rats. Only the least drug-like antagonist (3D53) maintained potency in cells against higher C5a concentrations and had a much longer duration of action (t1/2 ~ 20 h) than W54011 or JJ47 (t1/2 ~ 1 -3 h) in inhibiting macrophage responses. The unusually long residence time of 3D53 on its receptor was mechanistically probed by molecular dynamics simulations, which revealed long-lasting interactions that trap the antagonist within the receptor. Despite negligible oral bioavailability, 3D53 was much more orally efficacious than W54011 or JJ47 in preventing repeated agonist insults to induce rat paw oedema over 24 h. Thus, residence time on a receptor can trump drug-likeness in determining efficacy, even oral efficacy, of pharmacological agents.

No MeSH data available.


Related in: MedlinePlus