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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

3D53, W54011 and JJ47 are orally active anti-inflammatory compounds that inhibit C5a agonist-induced paw oedema in male Wistar rats.Paw oedema was induced in male Wistar rats by intraplantar administration of C5aR-PA (350 μg per paw in 100 μL of saline control). Paw swelling (% area change from baseline) was recorded at indicated timepoints. To determine comparative in vivo residence times, (A) 3D53 (5 mg/kg, p.o., n = 3 per group), (B) W54011 (30 mg/kg, p.o., n = 2 per group) and (C) JJ47 (30 mg/kg, p.o., n = 2 per group) were given orally 1 h prior to injection of C5aR-PA at specific time-points post-antagonism. Oral bioavailabilities of (D) 3D53 (F ~ 2%), (E) W54011 (F ~ 74%) and (F) JJ47 (F ~ 12%) were measured using LCMS to quantify plasma concentrations. C5aR antagonist was administering as an intravenous dose (1 mg/kg via an implanted jugular vein catheter in 50:50 DMSO/saline) versus an oral dose (10 mg/kg in olive oil, via gavage). Plasma was collected at various time points and analyzed. Error bars are means ± SEM normalized to maximal swelling of saline control. *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.
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f7: 3D53, W54011 and JJ47 are orally active anti-inflammatory compounds that inhibit C5a agonist-induced paw oedema in male Wistar rats.Paw oedema was induced in male Wistar rats by intraplantar administration of C5aR-PA (350 μg per paw in 100 μL of saline control). Paw swelling (% area change from baseline) was recorded at indicated timepoints. To determine comparative in vivo residence times, (A) 3D53 (5 mg/kg, p.o., n = 3 per group), (B) W54011 (30 mg/kg, p.o., n = 2 per group) and (C) JJ47 (30 mg/kg, p.o., n = 2 per group) were given orally 1 h prior to injection of C5aR-PA at specific time-points post-antagonism. Oral bioavailabilities of (D) 3D53 (F ~ 2%), (E) W54011 (F ~ 74%) and (F) JJ47 (F ~ 12%) were measured using LCMS to quantify plasma concentrations. C5aR antagonist was administering as an intravenous dose (1 mg/kg via an implanted jugular vein catheter in 50:50 DMSO/saline) versus an oral dose (10 mg/kg in olive oil, via gavage). Plasma was collected at various time points and analyzed. Error bars are means ± SEM normalized to maximal swelling of saline control. *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.

Mentions: Exogenous recombinant C5a protein is rapidly metabolized in serum within seconds-minutes to C5a-des Arg by carboxypeptidases, which remove the C-terminal arginine that is important for high affinity binding to C5aR28. Consequently, C5adesArg has 10–1000 times lower agonist potency than C5a depending upon the function being measured and cell type examined29. The very rapid degradation of recombinant C5a protein precludes its use in vivo as an agonist. Since only 6–8 residues at the C-terminus of C5a are responsible for agonist activity, although the remainder contribute to high affinity binding, a hexapeptide derivative of the C-terminus (Ac-FKP-dChaCha-dR-OH, referred to as C5aR-PA) was instead used for inducing C5aR-mediated paw oedema in rats. This hexapeptide is a more stable agonist than C5a but with a comparable spectrum of functional C5aR-mediated responses1730. This enabled comparison of the specific C5aR antagonists 3D53, W54011 and JJ47 for their effectiveness in inhibiting paw oedema induced by C5aR activation in vivo (Fig. 7).


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)

3D53, W54011 and JJ47 are orally active anti-inflammatory compounds that inhibit C5a agonist-induced paw oedema in male Wistar rats.Paw oedema was induced in male Wistar rats by intraplantar administration of C5aR-PA (350 μg per paw in 100 μL of saline control). Paw swelling (% area change from baseline) was recorded at indicated timepoints. To determine comparative in vivo residence times, (A) 3D53 (5 mg/kg, p.o., n = 3 per group), (B) W54011 (30 mg/kg, p.o., n = 2 per group) and (C) JJ47 (30 mg/kg, p.o., n = 2 per group) were given orally 1 h prior to injection of C5aR-PA at specific time-points post-antagonism. Oral bioavailabilities of (D) 3D53 (F ~ 2%), (E) W54011 (F ~ 74%) and (F) JJ47 (F ~ 12%) were measured using LCMS to quantify plasma concentrations. C5aR antagonist was administering as an intravenous dose (1 mg/kg via an implanted jugular vein catheter in 50:50 DMSO/saline) versus an oral dose (10 mg/kg in olive oil, via gavage). Plasma was collected at various time points and analyzed. Error bars are means ± SEM normalized to maximal swelling of saline control. *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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getmorefigures.php?uid=PMC4837355&req=5

f7: 3D53, W54011 and JJ47 are orally active anti-inflammatory compounds that inhibit C5a agonist-induced paw oedema in male Wistar rats.Paw oedema was induced in male Wistar rats by intraplantar administration of C5aR-PA (350 μg per paw in 100 μL of saline control). Paw swelling (% area change from baseline) was recorded at indicated timepoints. To determine comparative in vivo residence times, (A) 3D53 (5 mg/kg, p.o., n = 3 per group), (B) W54011 (30 mg/kg, p.o., n = 2 per group) and (C) JJ47 (30 mg/kg, p.o., n = 2 per group) were given orally 1 h prior to injection of C5aR-PA at specific time-points post-antagonism. Oral bioavailabilities of (D) 3D53 (F ~ 2%), (E) W54011 (F ~ 74%) and (F) JJ47 (F ~ 12%) were measured using LCMS to quantify plasma concentrations. C5aR antagonist was administering as an intravenous dose (1 mg/kg via an implanted jugular vein catheter in 50:50 DMSO/saline) versus an oral dose (10 mg/kg in olive oil, via gavage). Plasma was collected at various time points and analyzed. Error bars are means ± SEM normalized to maximal swelling of saline control. *p < 0.05, **p < 0.01 and ***p < 0.001 by student t-test.
Mentions: Exogenous recombinant C5a protein is rapidly metabolized in serum within seconds-minutes to C5a-des Arg by carboxypeptidases, which remove the C-terminal arginine that is important for high affinity binding to C5aR28. Consequently, C5adesArg has 10–1000 times lower agonist potency than C5a depending upon the function being measured and cell type examined29. The very rapid degradation of recombinant C5a protein precludes its use in vivo as an agonist. Since only 6–8 residues at the C-terminus of C5a are responsible for agonist activity, although the remainder contribute to high affinity binding, a hexapeptide derivative of the C-terminus (Ac-FKP-dChaCha-dR-OH, referred to as C5aR-PA) was instead used for inducing C5aR-mediated paw oedema in rats. This hexapeptide is a more stable agonist than C5a but with a comparable spectrum of functional C5aR-mediated responses1730. This enabled comparison of the specific C5aR antagonists 3D53, W54011 and JJ47 for their effectiveness in inhibiting paw oedema induced by C5aR activation in vivo (Fig. 7).

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