Isaac Cohen, O.M.D., Ph.D. November 6, 2015

Transcription

1 Isaac Cohen,.M.D., Ph.D. November 6, 15

2 Disclosure President and CE of Silurian Pharmaceuticals Share holder

3 Karenia brevis & Brevenal for Lung Diseases Karenia brevis is a marine microorganism K. brevis produces novel polyketide compounds Brevenal discovered in all Karenia brevis isolates Brevenal is an inhaled compound for the relief of bronchoconstriction and mucus clearance

4 Cystic Fibrosis & Neutrophil Elastase Davis SD, Ferkol T. N Engl J Med 13;368: Neutrophil proteases are critical for killing bacteria in the airways Accumulation of neutrophil elastase (NE) degrades structural proteins in airways This leads to bronchiectasis and bronchomalacia NE stimulates inflammatory response from airways epithelium NE interferes with bacterial clearance, impairing ciliary function and IgG NE cleaves chemokine and complement receptors

5 Brevenal Inhibits Neutrophil Elastase Induced Bronchoconstriction In Vivo Brevenal is effective >8hrs following HNE induced bronchoconstriction 7% Hypertonic saline has mild effect Effect of different agents on HNE-induced bronchoconstriction 1 Steroids (budesonide) has no effect on HNE induced bronchoconstriction Brevenal is superior to the elastase inhibitor Silvelestat Sodium Time course of 5 ug/ml brevenal on HNE-induced bronchoconstriction RL (% Change) HNE brevenal 7% HS Bud Sivelestat Na Values are mean + se for 6-9 sheep. Agents were given as aerosols 3 min -1h before HNE challenge. Brevenal ( breaths 5 ug/ml), Bud (budesonide 1 mg/3 ml), Sivelestate Sodium (1 mg ml). RL (% Change) Comparison of the HNE-inhibitor Sivelestat Sodium and brevenal on HNE-induced bronchoconstriction when given 4h before HNE challenge RL (% Change) 1 1 HNE.5h 4h 8h 24h Values are mean + se for 3-9 sheep. A single treatment ( breaths of 5 ug/ml brevenal) loses its effect against HNE between 8 and 24h. HNE brevenal Silevestat Na Values are mean + se for 3-9 sheep.

6 HNE caused a significant increase in total cells, neutrophils and macrophages TTAL CELLS * Post/Pre Challenge TC * TTAL CELLS * nly brevenal and S Na reduced the total cell response. Brevenal, Bud and S Na reduced the neutrophil response. CELLS x 1 3 / ml BAL 1 1 NEUTRPHILS * Post/Pre Challenge PMNs CNTRL BREV HS BUD AMIL S Na NEUTRPHILS * * * CNTRL BREV HS BUD AMIL S Na 5 There were no differences in the macrophage response MACRPHAGES * Post/Pre Challenge Macs MACRPHAGES Median values within box, lower box limit 25%, upper box limit 75%, upper wisker 9%, lower wisker 1%, data points outside these ranges are also plotted * P<.5 vs. BSL or CNTRL (n=8-26). BSL P-HNE CNTRL BREV HS BUD AMIL S Na TREATMENTS

7 Brevenal prevents WBC and Neutrophil Infiltration in Airways Agent RL TMV Total Cells Neutrophils HNE Control 119% 52% brevenal 24% 76% HS 126% 53% budesonide 77% 56% amiloride 76% 73% Mean pulmonary flow resistance (RL) Tracheal mucus velocity (TMV) Bronchoalveolar lavage (BAL) cell responses in conscious sheep after aerosol challenge with HNE Aerosols of hypertonic saline (HS, 3mL 7%) Glucocorticosteroid budesonide (1mg/3 ml, b.i.d for 4 days) Amiloride (3mL, 3mM) Brevenal ( breaths of 5ug/mL),

8 Effect of Various Treatments on CFTR (inh) -172 Slowing of Tracheal Mucus Velocity 11 TMV (% Baseline) h 1h 2h 3h 4h 5h 6h 7h 8h TIME CFTR(inh)-172 CFTR(inh) SNa CFTR(inh) HS CFTR(inh) Amiloride

9 Brevenal Reverses Effect of CFTR Inhibitor 172 In Vivo Effect of Brevenal on CFTR inh Induced Reduction in Tracheal Mucus Velocity 11 TMV (% of Baseline) 9 7 brevenal 5 h 1h 2h 3h 4h 5h 6h 7h 8h TIME CFTR (inh)-172 Vehicle (n=4, x + SD) CFTR (inh)-172 (n=3, x + SD) CFTR (inh)-172+brevenal (n=3, x + SD)

10 Comparative Time Course Effects of Brevenal and Amiloride on CFTR(inh)-172 Induced Slowing of Tracheal Mucus Velocity 1 11 TMV (% Baseline) h 1h 2h 3h 4h 5h 6h 7h 8h 9h 1h 11h 12h TIME CFTR(inh)-172 CFTR(inh) Amiloride CFTR(inh) Brevenal

11 Effect of CFTR (inh) -172 and HNE on Tracheal Mucus Velocity 11 TMV (% Baseline) 9 7 HNE Treatment 5 h 1h 2h 3h 4h 5h 6h 7h 8h 9h 1h 11h 12h TIME CFTR (inh) HNE CFTR (inh) HNE + Brevenal CFTR (inh) HNE + Albuterol

12 Brevenal Reverses CFTR (inh) High Dose HNE-Induced Reduction in TMV Single dose of Brevenal is effective for 24 hrs following inhibition of MCC by CFTR(inh) 172 plus high dose of HNE TMV (% Baseline) HNE Brevenal 5 h 1h 2h 3h 4h 5h 6h 7h 8h 9h 1h 11h 12h 24h Time CFTR (inh) LD HNE CFTR (inh) HD HNE CFTR (inh) HD HNE + Brevenal

13 Effect of Albuterol on HNE - Induced B ronchoconstriction Effect of Brevenal on HNE - Induced Broncoconstriction RL(% Change) RL (% Change) HNE ALB+HNE HNE Brevenal + HNE Effect Albuterol on C FT R(inh) + HNE - Induced B ronchoconstriction Effect of Brevenal on CFTR(inh) + HNE - Induced Bronchonstriction RL (% Change) RL (% Change) D1 D2 D3 D4 D1 D2 D3 D4

14 Brevenal Dose Response on ASL- Donor Prima

15 Brevenal a Voltage Sensitive Sodium Channel Inhibitor Brevenal VSSC inhibitor, binding to site 5 of VSSC brevenal brevetoxin Graph 1: Displacement Experiments for 3 H-PbTx-3 (1.5 nm) vs. 4 competitors Graph 1: Displacement Experiments for 3 H-Brevenol (8.5 nm) vs. 4 competitors % Total Binding Tritiated PbTx log [cold ligand] CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 H H 3 C PbTx-2 PbTx-3 Brevenal Brevenol H H H % Total Binding Tritiated Brevenol log [cold ligand] CH 3H H H H H H H 3 C H CH 3 R H H CH 3 H H brevenal R= C 38H 8 brevenol R= C 39H 62 8 reduced brevenol R= C 39H 66 8 H H H H CH 3 H 3C CH 3 H 3C CH 3 H 3C PbTx-2 PbTx-3 Brevenal Brevenol

16 Effectiveness Summary Brevenal is superior to HS, budesonide, amiloride, Silvelestat Sodium (an elastase inhibitor) and albuterol HNE induced changes in TMV HNE induced BC HNE induced inflammation Superior duration of action Brevenal reverses MCC inhibition caused by the CFTRinh 172 Brevenal reverses MCC inhibition caused by single and/or multiple challenges with CFTRinh HNE Brevenal increases ASL in primary donor cells Brevenal blocks TGFb induced ASL absorbtion

17 Contributors University of Miami- Mt. Sinai Medical Center W Abraham, PhD M Salathe, MD, PhD J R Sabater, MD University of North-Carolina, Wilmington D Baden, PhD A Bourdelais, PhD Stanford University C Milla, MD University of California, Berkeley Dale Leitman, MD, PhD CS Yuan, PhD