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Potent anti-inflammatory activity of novel microtubule-modulating brominated noscapine analogs.

Zughaier S, Karna P, Stephens D, Aneja R - PLoS ONE (2010)

Bottom Line: Noscapine, a plant-derived, non-toxic, over-the-counter antitussive alkaloid has tubulin-binding properties.Brominated noscapine analogs were found to inhibit cytokine and chemokine release from macrophage cell lines but did not affect cell viability.This study is the first report to identify brominated noscapines as innate immune pathway anti-inflammatory molecules.

View Article: PubMed Central - PubMed

Affiliation: Division of Infectious Diseases, Emory University School of Medicine, Atlanta, Georgia, United States of America. szughai@emory.edu

ABSTRACT
Noscapine, a plant-derived, non-toxic, over-the-counter antitussive alkaloid has tubulin-binding properties. Based upon the structural resemblance of noscapine to colchicine, a tubulin-binding anti-inflammatory drug, noscapine and its semi-synthetic brominated analogs were examined for in vitro anti-inflammatory activity. Brominated noscapine analogs were found to inhibit cytokine and chemokine release from macrophage cell lines but did not affect cell viability. Brominated noscapine analogs demonstrated anti-inflammatory properties in both TLR- and non-TLR induced in vitro innate immune pathway inflammation models, mimicking septic and sterile infection respectively. In addition, electron microscopy and immunoblotting data indicated that these analogs induced robust autophagy in human macrophages. This study is the first report to identify brominated noscapines as innate immune pathway anti-inflammatory molecules.

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Related in: MedlinePlus

Brominated noscapine analogs dampen TLR-mediated IL-8 release.HEK293 cells stably transfected with human TLR4-MD-2-CD14 receptors were co-treated with 50 µM of noscapine analogs and LPS concentrations ranging from 10-0.31 pM and incubated overnight. IL-8 release in supernatants was determined by ELISA. No drug: RAW264.7 cells treated with DMSO (50 µM) alone followed by LPS or Pam3CSK4 induction. Nos: noscapine and the TLR ligand. Br-nos: 9-bromonoscapine and the TLR ligand. Red-Br-nos: Reduced bromonoscapine and the TLR ligand. Unstimulated cells were used as control and were treated either with DMSO only or Br-nos (50 µM) without the TLR ligand. p values were calculated with reference to no drug values and were <0.0001 for Nos, Br-nos and Red-Br-nos drug treatment.
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pone-0009165-g006: Brominated noscapine analogs dampen TLR-mediated IL-8 release.HEK293 cells stably transfected with human TLR4-MD-2-CD14 receptors were co-treated with 50 µM of noscapine analogs and LPS concentrations ranging from 10-0.31 pM and incubated overnight. IL-8 release in supernatants was determined by ELISA. No drug: RAW264.7 cells treated with DMSO (50 µM) alone followed by LPS or Pam3CSK4 induction. Nos: noscapine and the TLR ligand. Br-nos: 9-bromonoscapine and the TLR ligand. Red-Br-nos: Reduced bromonoscapine and the TLR ligand. Unstimulated cells were used as control and were treated either with DMSO only or Br-nos (50 µM) without the TLR ligand. p values were calculated with reference to no drug values and were <0.0001 for Nos, Br-nos and Red-Br-nos drug treatment.

Mentions: To confirm the anti-inflammatory effects of brominated noscapine analogs, HEK293 human epithelial cells stably-transfected with TLR4-MD-2-CD14 receptors were co-treated with noscapine analogs (50 µM) and varying LPS concentration followed by overnight incubation. Brominated noscapine analogs significantly reduced TLR4-MD-2 mediated IL-8 release from HEK/TLR4-MD-2-CD14 cells (Figure 6). These drugs also significantly reduced IL-8 release by ∼3 fold (no drug = 1400 pg/ml, Br-nos = 650 pg/ml, Red-Br-nos = 400 pg/ml) from HEK293 cells transfected with TLR2 alone or TLR2/6 receptors induced with Pam3CSK4. Brominated noscapine analogs decreased inflammatory responses induced by both LPS (a TLR4-MD-2 ligand) and by Pam3CSK4 (a TLR2 ligand) suggesting that brominated noscapine analogs did not exert their anti-inflammatory effect by selectively inhibiting TLR4 and TLR2 cell surface receptor dimerization. Unlike paclitaxel that binds human and murine MD-2 [24], the anti-inflammatory role of brominated noscapines appeared to be due to effects on tubulin dynamics.


Potent anti-inflammatory activity of novel microtubule-modulating brominated noscapine analogs.

Zughaier S, Karna P, Stephens D, Aneja R - PLoS ONE (2010)

Brominated noscapine analogs dampen TLR-mediated IL-8 release.HEK293 cells stably transfected with human TLR4-MD-2-CD14 receptors were co-treated with 50 µM of noscapine analogs and LPS concentrations ranging from 10-0.31 pM and incubated overnight. IL-8 release in supernatants was determined by ELISA. No drug: RAW264.7 cells treated with DMSO (50 µM) alone followed by LPS or Pam3CSK4 induction. Nos: noscapine and the TLR ligand. Br-nos: 9-bromonoscapine and the TLR ligand. Red-Br-nos: Reduced bromonoscapine and the TLR ligand. Unstimulated cells were used as control and were treated either with DMSO only or Br-nos (50 µM) without the TLR ligand. p values were calculated with reference to no drug values and were <0.0001 for Nos, Br-nos and Red-Br-nos drug treatment.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2820095&req=5

pone-0009165-g006: Brominated noscapine analogs dampen TLR-mediated IL-8 release.HEK293 cells stably transfected with human TLR4-MD-2-CD14 receptors were co-treated with 50 µM of noscapine analogs and LPS concentrations ranging from 10-0.31 pM and incubated overnight. IL-8 release in supernatants was determined by ELISA. No drug: RAW264.7 cells treated with DMSO (50 µM) alone followed by LPS or Pam3CSK4 induction. Nos: noscapine and the TLR ligand. Br-nos: 9-bromonoscapine and the TLR ligand. Red-Br-nos: Reduced bromonoscapine and the TLR ligand. Unstimulated cells were used as control and were treated either with DMSO only or Br-nos (50 µM) without the TLR ligand. p values were calculated with reference to no drug values and were <0.0001 for Nos, Br-nos and Red-Br-nos drug treatment.
Mentions: To confirm the anti-inflammatory effects of brominated noscapine analogs, HEK293 human epithelial cells stably-transfected with TLR4-MD-2-CD14 receptors were co-treated with noscapine analogs (50 µM) and varying LPS concentration followed by overnight incubation. Brominated noscapine analogs significantly reduced TLR4-MD-2 mediated IL-8 release from HEK/TLR4-MD-2-CD14 cells (Figure 6). These drugs also significantly reduced IL-8 release by ∼3 fold (no drug = 1400 pg/ml, Br-nos = 650 pg/ml, Red-Br-nos = 400 pg/ml) from HEK293 cells transfected with TLR2 alone or TLR2/6 receptors induced with Pam3CSK4. Brominated noscapine analogs decreased inflammatory responses induced by both LPS (a TLR4-MD-2 ligand) and by Pam3CSK4 (a TLR2 ligand) suggesting that brominated noscapine analogs did not exert their anti-inflammatory effect by selectively inhibiting TLR4 and TLR2 cell surface receptor dimerization. Unlike paclitaxel that binds human and murine MD-2 [24], the anti-inflammatory role of brominated noscapines appeared to be due to effects on tubulin dynamics.

Bottom Line: Noscapine, a plant-derived, non-toxic, over-the-counter antitussive alkaloid has tubulin-binding properties.Brominated noscapine analogs were found to inhibit cytokine and chemokine release from macrophage cell lines but did not affect cell viability.This study is the first report to identify brominated noscapines as innate immune pathway anti-inflammatory molecules.

View Article: PubMed Central - PubMed

Affiliation: Division of Infectious Diseases, Emory University School of Medicine, Atlanta, Georgia, United States of America. szughai@emory.edu

ABSTRACT
Noscapine, a plant-derived, non-toxic, over-the-counter antitussive alkaloid has tubulin-binding properties. Based upon the structural resemblance of noscapine to colchicine, a tubulin-binding anti-inflammatory drug, noscapine and its semi-synthetic brominated analogs were examined for in vitro anti-inflammatory activity. Brominated noscapine analogs were found to inhibit cytokine and chemokine release from macrophage cell lines but did not affect cell viability. Brominated noscapine analogs demonstrated anti-inflammatory properties in both TLR- and non-TLR induced in vitro innate immune pathway inflammation models, mimicking septic and sterile infection respectively. In addition, electron microscopy and immunoblotting data indicated that these analogs induced robust autophagy in human macrophages. This study is the first report to identify brominated noscapines as innate immune pathway anti-inflammatory molecules.

Show MeSH
Related in: MedlinePlus