Limits...
The long-acting β2 -adrenoceptor agonist olodaterol attenuates pulmonary inflammation.

Wex E, Kollak I, Duechs MJ, Naline E, Wollin L, Devillier P - Br. J. Pharmacol. (2015)

Bottom Line: Besides their bronchodilatory effect, several studies suggest inhibitory effects on various aspects of inflammation.These anti-inflammatory effects were observed at doses relevant to their bronchodilatory efficacy.Furthermore, the in vivo data suggest that the anti-inflammatory properties of olodaterol are maintained after repeated dosing for 4 days.

View Article: PubMed Central - PubMed

Affiliation: Respiratory Diseases Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.

No MeSH data available.


Related in: MedlinePlus

Effect of olodaterol on CD11b expression, TNF-α release and neutrophil migration. (A) CD11b expression on granulocytes was measured after incubation of human peripheral whole blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 1 h in the presence of the drug. CD11b expression was determined by FACS using a monoclonal mouse anti-human PE-labelled CD11b/Mac-1 antibody. (B) TNF-α release was measured after incubation of blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 4 h in the presence of the drug. To test the antagonist effects, whole blood was pre-incubated with either ICI-118,551 (open triangles) or CGP-20712A (open squares) for 30 min before the addition of olodaterol (filled circles). (C) Transwell migration of isolated human peripheral blood neutrophils stimulated by different concentrations of IL-8. The number of migrated cells was determined using the CellTiter-Glo luminescent cell viability assay. Generation of the luminescent signal is proportional to the amount of ATP present, which signals the presence of metabolically active cells in the lower compartment. (D) Cells were pre-incubated with olodaterol (filled circles) or the CXCR2 antagonist SCH-527123 (open squares) for 30 min. Migration was stimulated by addition of 3 nM IL-8 to the lower chamber of the transwell. Data shown are means ± SEM (n = 3–4 donors).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4507158&req=5

fig06: Effect of olodaterol on CD11b expression, TNF-α release and neutrophil migration. (A) CD11b expression on granulocytes was measured after incubation of human peripheral whole blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 1 h in the presence of the drug. CD11b expression was determined by FACS using a monoclonal mouse anti-human PE-labelled CD11b/Mac-1 antibody. (B) TNF-α release was measured after incubation of blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 4 h in the presence of the drug. To test the antagonist effects, whole blood was pre-incubated with either ICI-118,551 (open triangles) or CGP-20712A (open squares) for 30 min before the addition of olodaterol (filled circles). (C) Transwell migration of isolated human peripheral blood neutrophils stimulated by different concentrations of IL-8. The number of migrated cells was determined using the CellTiter-Glo luminescent cell viability assay. Generation of the luminescent signal is proportional to the amount of ATP present, which signals the presence of metabolically active cells in the lower compartment. (D) Cells were pre-incubated with olodaterol (filled circles) or the CXCR2 antagonist SCH-527123 (open squares) for 30 min. Migration was stimulated by addition of 3 nM IL-8 to the lower chamber of the transwell. Data shown are means ± SEM (n = 3–4 donors).

Mentions: Stimulation of human whole blood with LPS concentration dependently induced the expression of CD11b on granulocytes and the release of TNF-α into the medium (data not shown). Olodaterol significantly attenuated the LPS-induced CD11b expression with a maximal inhibition of 48% at 10−8 M and a half-maximal inhibitory concentration (IC50) value of 1.31 × 10−10 M (Figure 6A) and almost completely blocked the TNF-α release with an IC50 value of 1.99 × 10−11 M (Figure 6B). In both assays, addition of the selective β2-adrenoceptor antagonist ICI-118,551 shifted the olodaterol curve to the right (133- and 168-fold respectively), whereas the selective β1-adrenoceptor antagonist CGP-20712A had no effect on neither the potency nor the maximal efficacy of olodaterol in both assays. In a transwell migration assay, IL-8 induced the migration of primary human neutrophils isolated from peripheral blood with a bell-shaped concentration dependence and a maximum at 3 nM IL-8 (Figure 6C). Olodaterol up to the highest concentration of 10−7 M showed no effect on the migratory capacity induced by 3 nM IL-8 (Figure 6D). To make sure that the lack of efficacy of olodaterol was not due to the lack of expression of functional β2-adrenoceptors on human neutrophils, a cAMP assay was performed, demonstrating a concentration-dependent increase in cAMP with olodaterol treatment (data not shown). Furthermore, to validate the transwell migration assay, the efficacy of the CXCR2 antagonist SCH-527123 was tested. SCH-527123 exhibited a potent and almost full inhibition of the IL-8-induced neutrophil migration (Figure 6D).


The long-acting β2 -adrenoceptor agonist olodaterol attenuates pulmonary inflammation.

Wex E, Kollak I, Duechs MJ, Naline E, Wollin L, Devillier P - Br. J. Pharmacol. (2015)

Effect of olodaterol on CD11b expression, TNF-α release and neutrophil migration. (A) CD11b expression on granulocytes was measured after incubation of human peripheral whole blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 1 h in the presence of the drug. CD11b expression was determined by FACS using a monoclonal mouse anti-human PE-labelled CD11b/Mac-1 antibody. (B) TNF-α release was measured after incubation of blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 4 h in the presence of the drug. To test the antagonist effects, whole blood was pre-incubated with either ICI-118,551 (open triangles) or CGP-20712A (open squares) for 30 min before the addition of olodaterol (filled circles). (C) Transwell migration of isolated human peripheral blood neutrophils stimulated by different concentrations of IL-8. The number of migrated cells was determined using the CellTiter-Glo luminescent cell viability assay. Generation of the luminescent signal is proportional to the amount of ATP present, which signals the presence of metabolically active cells in the lower compartment. (D) Cells were pre-incubated with olodaterol (filled circles) or the CXCR2 antagonist SCH-527123 (open squares) for 30 min. Migration was stimulated by addition of 3 nM IL-8 to the lower chamber of the transwell. Data shown are means ± SEM (n = 3–4 donors).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4507158&req=5

fig06: Effect of olodaterol on CD11b expression, TNF-α release and neutrophil migration. (A) CD11b expression on granulocytes was measured after incubation of human peripheral whole blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 1 h in the presence of the drug. CD11b expression was determined by FACS using a monoclonal mouse anti-human PE-labelled CD11b/Mac-1 antibody. (B) TNF-α release was measured after incubation of blood with olodaterol for 30 min and subsequently stimulated with 400 pg·mL−1 LPS for 4 h in the presence of the drug. To test the antagonist effects, whole blood was pre-incubated with either ICI-118,551 (open triangles) or CGP-20712A (open squares) for 30 min before the addition of olodaterol (filled circles). (C) Transwell migration of isolated human peripheral blood neutrophils stimulated by different concentrations of IL-8. The number of migrated cells was determined using the CellTiter-Glo luminescent cell viability assay. Generation of the luminescent signal is proportional to the amount of ATP present, which signals the presence of metabolically active cells in the lower compartment. (D) Cells were pre-incubated with olodaterol (filled circles) or the CXCR2 antagonist SCH-527123 (open squares) for 30 min. Migration was stimulated by addition of 3 nM IL-8 to the lower chamber of the transwell. Data shown are means ± SEM (n = 3–4 donors).
Mentions: Stimulation of human whole blood with LPS concentration dependently induced the expression of CD11b on granulocytes and the release of TNF-α into the medium (data not shown). Olodaterol significantly attenuated the LPS-induced CD11b expression with a maximal inhibition of 48% at 10−8 M and a half-maximal inhibitory concentration (IC50) value of 1.31 × 10−10 M (Figure 6A) and almost completely blocked the TNF-α release with an IC50 value of 1.99 × 10−11 M (Figure 6B). In both assays, addition of the selective β2-adrenoceptor antagonist ICI-118,551 shifted the olodaterol curve to the right (133- and 168-fold respectively), whereas the selective β1-adrenoceptor antagonist CGP-20712A had no effect on neither the potency nor the maximal efficacy of olodaterol in both assays. In a transwell migration assay, IL-8 induced the migration of primary human neutrophils isolated from peripheral blood with a bell-shaped concentration dependence and a maximum at 3 nM IL-8 (Figure 6C). Olodaterol up to the highest concentration of 10−7 M showed no effect on the migratory capacity induced by 3 nM IL-8 (Figure 6D). To make sure that the lack of efficacy of olodaterol was not due to the lack of expression of functional β2-adrenoceptors on human neutrophils, a cAMP assay was performed, demonstrating a concentration-dependent increase in cAMP with olodaterol treatment (data not shown). Furthermore, to validate the transwell migration assay, the efficacy of the CXCR2 antagonist SCH-527123 was tested. SCH-527123 exhibited a potent and almost full inhibition of the IL-8-induced neutrophil migration (Figure 6D).

Bottom Line: Besides their bronchodilatory effect, several studies suggest inhibitory effects on various aspects of inflammation.These anti-inflammatory effects were observed at doses relevant to their bronchodilatory efficacy.Furthermore, the in vivo data suggest that the anti-inflammatory properties of olodaterol are maintained after repeated dosing for 4 days.

View Article: PubMed Central - PubMed

Affiliation: Respiratory Diseases Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.

No MeSH data available.


Related in: MedlinePlus