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The Effect of PUR1800, a Novel Narrow Spectrum Kinase Inhibitor, on Viral Replication and Viral-Induced Inflammation in Primary Human Airway Cells
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A4774 - The Effect of PUR1800, a Novel Narrow Spectrum Kinase Inhibitor, on Viral Replication and Viral-Induced Inflammation in Primary Human Airway Cells
Author Block: A. Curran1, K. Ito2, D. Hava1, C. Charron2; 1Pulmatrix, Lexington, MA, United States, 2RespiVert Ltd, London, United Kingdom.
PUR1800 (RV1162) is a novel narrow-spectrum kinase inhibitor (NSKI) that inhibits p38 MAPK, src and syk kinases. To test the activity of this compound against viral replication and viral-induced inflammation, studies were performed in cultured primary human bronchial epithelial cells (HBEC) from both healthy human volunteers (HHV) and COPD patients.
HBECs were cultured from HHVs and infected with Respiratory Syncytial Virus (RSV) strain A2 for 1h and cells incubated for 4 days. HBECs were treated 2h prior to infection and 1h after viral washout with a DMSO control, a dose range of PUR1800 or a single high dose of ribavirin (10μg/mL) as a positive control. After 4 days of incubation, viral load was assessed by F Protein expression as detected by ELISA. Ribavirin inhibited RSV-F protein expression by 94.5%, with 61% cell viability relative to the control. PUR1800 (0.04μg/mL) maximally inhibited RSV-F protein expression by 89.6% with >90% cell viability and an IC50 of 10.9nM (6.4 ng/mL).
In a separate study, HBECs were cultured from HHVs and COPD patients in an air-liquid interface culture system. Cells were stimulated for 2h with poly(I:C), an agonist of TLR3, a receptor that stimulates inflammatory responses to viral exposure. HBECs were treated during poly(I:C) stimulation with vehicle, PUR1800 (0.01 or 0.1μg/mL) or fluticasone propionate (0.01 or 0.1μg/mL) and cells were incubated for 24h. A Luminex Multiplex assay (Millipore) was used to assay IL-6, TNFα, CXCL8, Eotaxin, MIP-1β, G-CSF and GM-CSF compared to control levels. Poly (I:C) treatment similarly induced all cytokines in both HHV and COPD cells. PUR1800 reduced the expression of each cytokine in a concentration-dependent manner. In COPD cells, PUR1800 treatment resulted in 95.0% inhibition of IL-6, 88.7% inhibition of TNFα and 58.5% inhibition of CXCL8, and >50% inhibition of all cytokines assayed. Similar effects were observed in HHV cells. Fluticasone showed weak anti-inflammatory effects with no greater than 25% inhibition of any cytokine in either HHV or COPD cells.
Combined, these data demonstrate that PUR1800 (RV1162) inhibits viral replication and the inflammatory response to viral infection in human airway cells from both HHVs and COPD patients. These results suggest that PUR1800 shows promise as a treatment of viral-induced COPD exacerbations.
Author Block: A. Curran1, K. Ito2, D. Hava1, C. Charron2; 1Pulmatrix, Lexington, MA, United States, 2RespiVert Ltd, London, United Kingdom.
PUR1800 (RV1162) is a novel narrow-spectrum kinase inhibitor (NSKI) that inhibits p38 MAPK, src and syk kinases. To test the activity of this compound against viral replication and viral-induced inflammation, studies were performed in cultured primary human bronchial epithelial cells (HBEC) from both healthy human volunteers (HHV) and COPD patients.
HBECs were cultured from HHVs and infected with Respiratory Syncytial Virus (RSV) strain A2 for 1h and cells incubated for 4 days. HBECs were treated 2h prior to infection and 1h after viral washout with a DMSO control, a dose range of PUR1800 or a single high dose of ribavirin (10μg/mL) as a positive control. After 4 days of incubation, viral load was assessed by F Protein expression as detected by ELISA. Ribavirin inhibited RSV-F protein expression by 94.5%, with 61% cell viability relative to the control. PUR1800 (0.04μg/mL) maximally inhibited RSV-F protein expression by 89.6% with >90% cell viability and an IC50 of 10.9nM (6.4 ng/mL).
In a separate study, HBECs were cultured from HHVs and COPD patients in an air-liquid interface culture system. Cells were stimulated for 2h with poly(I:C), an agonist of TLR3, a receptor that stimulates inflammatory responses to viral exposure. HBECs were treated during poly(I:C) stimulation with vehicle, PUR1800 (0.01 or 0.1μg/mL) or fluticasone propionate (0.01 or 0.1μg/mL) and cells were incubated for 24h. A Luminex Multiplex assay (Millipore) was used to assay IL-6, TNFα, CXCL8, Eotaxin, MIP-1β, G-CSF and GM-CSF compared to control levels. Poly (I:C) treatment similarly induced all cytokines in both HHV and COPD cells. PUR1800 reduced the expression of each cytokine in a concentration-dependent manner. In COPD cells, PUR1800 treatment resulted in 95.0% inhibition of IL-6, 88.7% inhibition of TNFα and 58.5% inhibition of CXCL8, and >50% inhibition of all cytokines assayed. Similar effects were observed in HHV cells. Fluticasone showed weak anti-inflammatory effects with no greater than 25% inhibition of any cytokine in either HHV or COPD cells.
Combined, these data demonstrate that PUR1800 (RV1162) inhibits viral replication and the inflammatory response to viral infection in human airway cells from both HHVs and COPD patients. These results suggest that PUR1800 shows promise as a treatment of viral-induced COPD exacerbations.
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