CRTh2: Can Residence Time Help ?
RSC-SCI Cambridge Medicinal Chemistry Symposium
Churchill College, Cambridge 8-11 September 2013
Rick Roberts Almirall Introduction
Paul Ehrlich, The Lancet (1913), 182, 445
“A substance will not work unless it is bound”
100 years on:
“What a substance does may depend on how long it is bound”
2 In a nutshell ….
Introduction
“Applications of Binding Kinetics to Drug Discovery” D. Swinney, Pharm. Med. (2008), 22, 23-34
3 Energetic concept of Residence Time
kon Standard model [R] + [L] [RL] k unbound off bound
Energy
‡ Binding process (kon) is (R·L) determined by the energy
barrier Ea E Ligand concentration a Unbinding process (koff) is determines the number of Ed determined by the energy attempts to get over this barrier Ed barrier [R] + [L]
G = Ed -Ea
[RL] Reaction coordinate
Potency (Ki) is determined by the difference in these energies
koff Ki = kon
4 K , k , k i on off -1 -1 kon M s Potency (nM) vs kon vs koff Very fast association 108 10 1 0.1 0.01 0.001 107 100 10 1 0.1 0.01 0.001 fast association 106 1000 100 10 1 0.1 0.01 0.001 105 10 M 1000 100 10 1 0.1 0.01 0.001 slow association 104 10 M 1000 100 10 1 0.1 0.01 Very slow association 103 10 M 1000 100 10 1 0.1 “inactive” 102 10 M 1000 100 10 1 10 10-1 10-2 10-3 10-4 10-5 10-6 10-7 s-1 koff
Energy A simple binding measurement gives no clue to how fast the association and dissociation are (R·L)‡
Very slow associating compounds that are very slow dissociating (R·L)‡ slow associating compounds that are slow dissociating Are all fast associating compounds that are fast dissociating equipotent Very fast associating compounds that are very fast dissociating [R] + [L]
If we can control the transition state energy, we can control the binding kinetics [RL] Reaction coordinate
5 What is the transition state (R·L)‡ ? entropy Protein conformational Partial changes desolvations Energy-lowering steps to get to Fleeting final bound state existence
A physical process Partial electrostatic interactions
Solvated Solvated Calculate G and Ligand Receptor hence potency
A calculated process + interaction + interaction - 10 × from ligand + Van der Waals interaction - 6 × from receptor - Entropy of ligand +/- etc Water molecules - Molecular orbital orientation Molecular interaction points +/- etc
6 Why bother to control Binding Kinetics ? Residence time / Dissociation half-life seconds minutes hours days level PD
PK
time
Mechanistic Surmountable Insurmountable Pharmacodynamic
Potency has little influence over these behaviours
Partial agonism ↔ Full agonism
Functional efficacy relates to off-rate .M3 agonists Mol. Pharmacol. (2009), 76, 543-551
.A2A agonists Br. J. Pharmacol. (2012), 166, 1846-1859
7 Why bother to control Binding Kinetics ? Residence time / Dissociation half-life seconds minutes hours days level PD
PK
time
Mechanistic Surmountable Insurmountable Pharmacodynamic Efficacy depends on substrate concentration Enzyme inhibitors can cause a build-up of substrate .Lovastatin, an HMG-CoA reductase inhibitor is sensitive to HMG- CoA build-up .Candasartan, an ACE inhibitor is insensitive to AT1 build-up
Mechanism-based toxicity Dopamine D2 antagonists .Clozapine as a rapidly reversible D2 antagonist .Haloperidol is insurmountable and blocks basal D2 activity GI Toxicity of COX-1 inhibitors. Lett. Drug Design Disc. (2006), 3, 569 .Celecoxib is surmountable and free of GI ulceration .Aspirin is irreversible and carries use warnings
8 Why bother to control Binding Kinetics ? Residence time / Dissociation half-life seconds minutes hours days level PD
PK
time
Mechanistic Surmountable Insurmountable Pharmacodynamic
Pharmacodynamics outlasts Pharmacokinetics
Ester hydrolysable M3 antagonists .Ipratropium is dosed 2 puffs, 4 times a day .Aclidinium and Tiotropium are twice and once daily respectively
Kinetic selectivity M3 over M2 selectivity .Aclidinium bromide safety profile. J. Pharm. Exp. Ther (2009), 331, 740-751
9 On rates by mechanism – also independent of potency Simple fit Induced fit
kon k1 k3 [R] + [L] [RL] [R] + [L] [RL] [RL*]
koff k2 k4
. If the off rate is slow, . The first on and off rates are quicker the on rate is also slow . Some kind of internal change takes place once bound . The final off rate is macroscopically the slowest
Energy Energy
‡ (R·L)‡ (RL)
(R·L)‡ slow Ea Ed
[R] + [L] [R] + [L] quick [RL] [RL] [RL*] Reaction coordinate Reaction coordinate
10 On rates by mechanism Simple fit
kon [R] + [L] [RL]
koff
. If the off rate is slow, the on rate is also slow
Simple fit model Once “on”, the ligand behaves as an 100 insurmountable antagonist 80
However, the time to 60 “get on” is about 6 h. 40
Before this time the 20 ligand is a partial % Receptor Occupancy antagonist 0 6 121824 Time (h) Simulation: Dissociation half-life 24 h
Concentration of [L] at 10 × IC50
Sufficient PK levels needed for sufficient time to ensure receptor becomes saturated
11 On rates by mechanism
Induced fit
k1 k3 [R] + [L] [RL] [RL*]
k2 k4
The ligand binds quickly (Total RO%) However at this point it behaves as a classical surmountable ligand Induced fit model Once “on”, the ligand behaves as an 100 insurmountable antagonist 80 Total RO% 60 RL*% 40 RL% 20 % Receptor Occupancy 0 6 121824 Time (h)
Simulation: k4 Dissociation half-life 24 h Concentration of [L] at 10 × IC50
Sufficient PK levels needed for sufficient time to ensure receptor becomes saturated
12 The CRTh2 Programme
13 Brief introduction to CRTh2 Real name:
.Chemoattractant Receptor- homologous molecule expressed on T-Helper 2 cells
.Also known as DP2
CRTh2 activation:
.induces a reduction of intracellular cAMP and calcium mobilization.
.is involved in chemotaxis of Th2 lymphocytes, eosinophils, mast cells and basophils. CRTh2 and DP1 review. Nat. Rev. Drug Disc. (2007), 6, 313
.inhibits the apoptosis of Th2 lymphocytes CRTh2 antagonism:
.stimulates the production of IL4, IL5, and .Should block pro-inflammatory PGD2 effects on IL13, leading to: key cell types . eosinophil recruitment and survival . mucus secretion .Potential benefit in: . airway hyper-responsiveness . asthma . immunoglobulin E (IgE) production . allergic rhinitis . etc . atopic dermatitis.
14 Why Long Residence Time in CRTh2 ? Indole acetic acids Cl
HO O HO O N F N S S N O O NH ?
O Oxagen-Eleventa AstraZeneca Actelion Novartis Merck Pulmagen-Teijin
OC-459 AZD-1981 Setipiprant R = CF3 QAV-039 MK-7246 ADC-3680 Phase III Phase II Phase III R = H QAW-680 Phase I Phase II Phase II/II
Aryl acetic acids
? Indomethacin Weak agonist Boehringer Array Roche BI671800 ARRY-502 RG-7581 Phase II Phase II Phase I Phenoxyacetic acids HN N O
HO O CF3
OMe AstraZeneca Panmira Panmira Amgen AZD-5985 or AM461 AM211 AMG-853 AZD-8075 Phase I Phase I Phase II Phase I/I
15 PK of carboxylic acids PK Half-life vs Volume 10 2. Ideal CRTh2 Acid 3. zone Zwitterion 1 Human PK profiles of Total body water volume 150 carboxylic acids
0.1 Adapted from Obach, Total plasma volume Drug.Metab.Disp. 4. 1.
(2008), 36, 1385 Volume of distribution (L/kg) 0 6 12 18 24 Human terminal half-life (h) Techniques used to avoid rapid clearance:
1.Very high plasma protein binding (e.g. Diflunisal >99%)
2.Zwitterionic tissue distribution (e.g. Trovafloxacin, Vss 1.3 L/kg) 3.Organ distribution (e.g. Pravastatin, t½ 0.5 h, but concentrated in the liver) 4.Be small, be ignored by the liver (e.g. Probenecid, 80-90% renal excretion)
O O F O OH HO O H N N N OH OH O H F H N OH 2 H
F Diflunisal Trovafloxacin Pravastatin Probenecid PK t½ 10h PK t½ 11h PK t½ 0.5h PK t½ 5.9h
16 Our internal programme We want to find a once a day, oral, low dose (≤ 10 mg) CRTh2 antagonist for mild-moderate asthma
. All antagonists are acids.
. Acids generally have poor PK
. Clinical dosing of first CRTh2 antagonists – high dose and twice daily
. Therefore, we chose to deliberately look for slowly dissociating CRTH2 antagonists:
. to maintain receptor occupancy beyond normal PK and extend duration of action . to reduce the pressure of finding a carboxylic acid with desirable PK properties
. to add the possibility of extra protection due to insurmountability against PGD2 burst release
Residence time / Dissociation half-life seconds minutes hours days level PD
PK
time
Mechanistic Surmountable Insurmountable Pharmacodynamic
17 Are there slow-dissociating CRTh2 antagonists?
O
HO H N F N S O O Ramatroban TM-30642 TM-30643 TM-30089 (CAY-10471)
Structure not disclosed Merck Amira Pulmagen MK-7246 PGD2 AM432 ADC-3680
Dissociation Compound Potency Reference half-life*
Ramatroban pA2 36 nM 5 min Mol. Pharmacol. (2006), 69, 1441
TM-30642 pA2 20 nM 8 min --- / / ---
TM-30643 pA2 4 nM (12.8 years) --- / / ---
TM-30089 pA2 (13.5 years) --- / / ---
MK-7246 Ki 2.5 nM 33 min Mol. Pharmacol. (2011), 79, 69
PGD2 KD 11 nM 11 min Bioorg. Med. Chem. Lett. (2011), 21, 1036
AM432 IC50 6 nM 89 min --- / / ---
ADC-3680 Ki 1.6 nM 20 min American Thoracic Society (ATS), May 17-22, 2013
18 In vitro dissociation assays CRTh2 GTPS binding Compound PDG2 .Membranes over-expressing human CRTh2 35 3 h .PGD2 agonism produces allows [ S]-GTPS binding, detected by radioactivity
Ramatroban 0 → 10 µM Readout time 5000 Readout 4000 1 h 3000 Classical Surmountable 2000 Readout inhibition Ramatroban 1000
0 0.1 nM 10 nM 1 µM 100 µM
PGD2 Concentration 1000
800 Readout Classical 15 min 600 Insurmountable 400 inhibition Readout
TM-30089 200 (CAY-10471) 0 0.1 nM 10 nM 1 µM 100 µM
PGD2 Concentration CAY-10471 0 → 5 µM
19 In vitro dissociation assays CRTh2 Surmountability vs Insurmountability Compound PDG2 .Membranes over-expressing human CRTh2 35 3 h .PGD2 agonism produces allows [ S]GTPS binding, detected by radioactivity
Ramatroban 0 → 10 µM Readout time 24 h 5000 Readout 4000 1 h 3000 Classical Surmountable 2000 Readout inhibition Ramatroban 1000
0 0.1 nM 10 nM 1 µM 100 µM
PGD2 Concentration 1000 Re-mountable inhibition
800 Readout 6000 Readout 600 15 min 24 h
400 4000 Readout Readout TM-30089 200 (CAY-10471) 2000 0 0.1 nM 10 nM 1 µM 100 µM 0.1 nM 10 nM 1 µM 100 µM
Surmountability is just a PGD2 Concentration PGD2 Concentration question of time CAY-10471 0 → 5 µM
20 In vitro dissociation assays CRTh2 Washout experiments Compound PDG2 (5 × K ) (1000 × Ki) .Membranes pre-incubated with compound i .Antagonist effectively swamped by agonist 2 h .Decay of inhibition followed by time .Filtration reading in automated 96-well format .Readings from 2 min to 28 h Readout times
100
80
60 Dissociation t1/2 < 1 min
40
Ramatroban 20 “Worst case” scenario for Mean InhibitionMean % dissociation half-life Surmountable 0 (Competitive) 0 4 8 12 16 20 24 28 Time (h) .No rebinding possible. 100
80 .Physiological system may Dissociation t1/2 ~80 min be less demanding 60
40
20
TM-30089 Mean Inhibition % (CAY-10471) 0 0 4 8 12 16 20 24 28 Insurmountable Time (h) (Non-Competitive)
21 Pyrazoles and Pipas . First series of Pyrazoles gave active compounds, but no residence
. Indole nucleus developed by Oxagen. Beginnings of slow dissociation observed
. Pyrrolopiperidinones (Pipas) gave both activity and the first observations of long residence
* *
Pyrazoles Indole Pyrrolopiperidinones (Pipas)
* * * * * * N * N N * * * * * * * Core SAR N O O N N H
Core Pyrazole Reverse 5-F-Indole PiPa N-Me Pipa N-Bn Pipa diMe-Pipa
GTPS IC50 (nM) 7 35 14 5 5 1 4 Dissociation t½ (h) 0.1 0.1 1.3 2.3 5.3 6.6 10
All compounds of similar potency Core structure has a greater effect on Residence Time
22 Pipa tail SAR
. Substituent changes in the tail did not greatly affect the potency
. There is however an additive effect on residence time
1 2 R R GTPS IC50 Dissociation t½ H H 5 nM 2.3 h
MeO H 4 nM 4.2 h
H F 5 nM 3.3 h
MeO F 7 nM 23 h
. Pipa series was essentially impermeable.
. Not suitable for an oral program
. Still, a valuable tool to validate the PK-PD disconnection in guinea pig
23 PK-PD Disconnection Simulations At t = 0, [Ligand] = 10 × IC50 PK half-life 1h PK half-life 1h Dissociation half-life 1h Dissociation half-life 12h 100 100 Total RO% Total RO% 80 PK RO% 80 PK RO% GPCR 60 60 threshold Barely 40 observable 40
20 20 Observable % Receptor Occupancy % Receptor Occupancy 0 0 0 10 20 30 0 10 20 30 Time (h) Time (h) PK half-life 12h PK half-life 12h Dissociation half-life 1h Dissociation half-life 12h 100 100
80 80 GPCR 60 Not observable 60 Barely threshold observable 40 40 Total RO% PK RO% Total RO% 20 20 PK RO% % Receptor Occupancy % Receptor Occupancy 0 0 0 10 20 30 0 10 20 30 Time (h) Time (h) For a recent article, see Drug Disc. Today (2013), 18, 697
24 PK-PD disconnection model proof of principle Guinea Pig Eosinophilia assay
Compound DK-PDG2 .Guinea pigs are pre-dosed with test compound 1 - 24h .Wait desired time: 1h, 5h, 17h, 24h ….. 1h
.Anaesthetised animals dosed with i.v. DK-PGD2, .DK-PGD2 via CRTh2 causes liberation of Readout time eosinophils from bone marrow to plasma .After 1 h blood is extracted .Simultaneous eosinophil counting and PK analysis Accompanying PK analyses
Finding the right tool compounds
Human v Gp dissociation
30 (MeO,F)-Pipa F-indole (diMe)-Pipa 1. Species Dissociation half-lives 20 Compound Core human Guinea pig 3. 10 1 (MeO,F)-Pipa 23 h 2 h Human half-life (h) half-life Human 2. 2 F-Indole 1.4 h 1 h 10 20 30 Guinea Pig half-life (h) 3 (diMe)-Pipa 10 h 20 h
25 PK-PD disconnection model Indole gp PK 0.3 mg/kg p.o.
10000 Guinea pig 2h eosinophilia IC 1000 50
100
10 F-Indole guinea pig profile
2h Eosinophilia IC50 40 ng/ml 1 Dissociation t½ 1.3 h Plasma (ng/ml) conc 6 12 18 24 PK t½ 5.3 h 0.1 Time (h)
Series Dose Cmax (ng/ml) AUCinf (ng/ml/h) Cl/F (ml/min/kg) Vz/F (L/kg) t1/2 (h) Indole 0.3 mg/kg p.o. 481 4796 1 0.47 5.3
. Inhibition of eosinophilia (PD) is Indole Eosiniophilia PK-PD Pre-administration purely dependant upon systemic time levels (PK) 150
1 h 30 . No PK-PD disconnection 100 3 3 PK half-life 12h 24 h Dissociation half-life 1h 0.3 50 0.3 100 Dose mg/kg 80 0 60 0.03 10 100 1000 10000 100000 40 Total RO% % Inhibition Eosinophilia -50 20 PK RO% Plasma conc (ng/ml) % ReceptorOccupancy 0 0 10 20 30 Time (h)
26 PK-PD disconnection model Pipa gp PK 3 mg/kg s.c.
HO O 10000
N S Guinea pig 2h O 1000 O eosinophilia IC50 O 100 N H 10 diMe-Pipa guinea pig profile
2h Eosinophilia IC50 7 ng/ml 1 Dissociation t½ 20 h Plasma (ng/ml) conc 6 12 18 24 PK t½ 0.9 h 0.1 Time (h)
Series Dose Cmax (ng/ml) AUCinf (ng/ml/h) Cl/F (ml/min/kg) Vz/F (L/kg) t1/2 (h) Pipa 3 mg/kg s.c. 1633 2619 20 1.5 0.9
. Inhibition of eosinophilia (PD) Pre-administration Pipa Eosinophilia outlasts drop in systemic levels time (PK) at 17h after dosing 120 1 h 100 3 3 . PK-PD disconnection (hysteresis) 15 h 80 0.3 0.3
PK half-life 1h 60 Dissociation half-life 12h 40 0.03 100 Total RO%
80 PK RO% % inhibition eosinophilia 20 Dose mg/kg 0.003 60 0 0.1 1 10 100 1000 40 -20 20 plasma conc ng/ml % ReceptorOccupancy 0 0 10 20 30 Time (h)
27 How long is long residence ?
Saturate Receptor, then washout 1st half-life 2nd half-life 3rd half-life
Total receptor population
Ligand lost Too dilute to rebind Cleared from circulation
Receptor Occupancy by exponential decay 100
80 Receptor Occupancy required for a GPCR antagonist. 60 Pharmacol. Therap. (2009), 122, 281-301
40 This translates to about half a Dissociation Half-life 20
Receptor Occupancy To turn a twice-daily compound 0 into a once-daily compound, 0 132 4 6 8 10 Half-lives of dissociation we want a Dissociation Half-life of ≥ 24h
28 Where is long Residence? Selected lit reports or Structure Residence Relationships (SRRs)
.Trend analysis of D2 antagonists. Bioorg. Med. Chem (2011), 19, 2231. .Trend analysis of Pfizer and literature data. Med. Chem. Comm. (2012), 3, 449 .Review of molecular determinants. Drug Disc. Today. (2013), 18, 667
Are more active compounds longer resident ? Dissociation Half-life vs Potency Energy (R·L)‡ 1000 Long residence 1 – 100 nM Biaryl 100 PiPa Pyrazole 10 [R] + [L] Longer (nM) Potency Other 50 resident? 1 Std S IC “Non” resident 10 – 1000 nM [RL]
More GTP 0.1 potent Short Residence 8421 3216 [RL] Dissociation Half-life (h) Reaction coordinate
Intuitive, but not that helpful. We want the most potent compounds anyway
29 Where is long Residence?
Are more Polar compounds longer resident?
Is desolvation (R·L)‡ relevant here? Energy Dissociation Half-life vs TPSA (R·L)‡ 150 Biaryl Pyrazole
) Pipa 2 100 Å 6 Mem Competitor [R] + [L] Other
TPSA ( 50
[RL] 0 Short Residence 1 2 4 8 16 32 Reaction coordinate Dissociation Half-life (h)
Freedom to adapt polar surface area to modulate physicochemical properties
30 Where is long Residence?
Are Impermeable compounds are Long Resident in CRTh2 ?
ResTime vs Permeability 100
10 Biaryl cm/s) -6 1 Mono Other 0.1 Pyr PAMPA 0.01 RevPyr PiPa 0.001 Std
Permeability (x10 Permeability 0.0001 0.031250.06250.1250.25 0.5 1 2 4 8 16 32 Dissociation Half-life (h) Not Intuitive
31 Where is long Residence?
Are Impermeable compounds are Long Resident in CRTh2 ?
ResTime vs Permeability 100
10 Biaryl cm/s) -6 1 Mono Other 0.1 Pyr PAMPA 0.01 RevPyr PiPa 0.001 Std
Permeability (x10 Permeability 0.0001 0.031250.06250.1250.25 0.5 1 2 4 8 16 32 Dissociation Half-life (h) Not Intuitive And ultimately not true
Design permeable compounds for oral delivery
32 Where is long Residence?
Are more lipophilic compounds longer resident?
Energy
(R·L)‡ Dissociation Half-life vs cLogP
Biaryl [R] + [L] 6 Pyrazole Pipa 4 6 Mem [R] + [L] Competitor
cLogP 2 Other More potent [RL] 0 Short Residence 1 2 4 8 16 32 Reaction coordinate Dissociation Half-life (h) Probably just pushes up energy of [L] in free water Maybe a slight overall trend ?
Lipophilic molecules are often more potent, because once bound, the molecule doesn’t want to go back into the surrounding water
33 Where is long Residence?
Are more lipophilic compounds longer resident?
Within particular sub-series, there was often a relationship
O N
HO O 7-Azaindole Dissociation Half-life vs cLogP O IC50 N 21 nM O HN R N N Diss N 6 HO O R t½ 5 h R cLogP 4 -0.6 isoxazoles CF3-Indole cLogP 2 IC50 37 nM Diss 0 t½ Short Residence 1 2 4 8 16 32 27 h cLogP Dissociation Half-life (h) 5-fold increase in Dissociation5.9 half-life For a 1,000,000-fold increase in lipophilicity
Workable in final optimisation, but not without its associated risks for oral delivery
34 Where is long Residence?
Series by series First example of series Pyrazole Series 6-Mem Series Competitor Compounds 15 5 5
4 4 10 3 3
2 2 5 1 1 Number of Compounds Number of Compounds Number of Compounds 0 0 0 1 min 1 2 4 8 16 32 1 min 1 2 4 8 16 32 1 min 1 2 4 8 16 32 Dissociation Half-life (h) Dissociation Half-life (h) Dissociation Half-life (h) Pipa Series Biaryl Series Other compounds 10 60 10 HO O 8 Ar 8 N 40 6 6 O N 4 H 4 20 2 2 Number ofCompounds Number ofCompounds Number ofCompounds 0 0 0 1 min 1 2 4 8 16 32 1 min 1 2 4 8 16 32 1 min 1 2 4 8 16 32 Dissociation Half-life (h) Dissociation Half-life (h) Dissociation Half-life (h)
. Chemical series drives Residence Time: If you’ve got it, you’ve got it and if you don’t, you don’t . First compound of series dictates the trend.
35 Homing in on Residence Time in the Biaryl series
IC50 5 nM IC50 8 nM Dissociation Dissociation half-life half-life 9 h 11 h
S S HO O HO O O IC50 4 nM CF N 3 N inactive HN Dissociation
N N half-life 25 min
Long Residence Time in the Biaryls comes from an H-Bond Acceptor in ortho
36 Amide Rotamers
50:50 N O by NMR HO O CF3 cis trans IC50 14 nM OMe Dissociation half-life 2 h
inactive IC50 27 nM half-life 1.5 h
Long Residence Time in the Biaryls comes from an H-Bond Acceptor in ortho in a specific position
37 Can we check the H-Bond Acceptor location?
O N O N HO O HO O
N N
N R
1 R = H F 2
R= Me. Ab initio calculations conformational preference
Compound 1 2
GTPS IC50 (nM) 20 6 Dissociation t½ (h) 10 18
38 Can we achieve truly long residence?
Position-by-position analysis of good structural features for Residence Time
Benzyl to bump up lipophilicity
Isoxazole as H-bond acceptor Acetic acid
Chloro is OK here 7-Azaindole for phys-chem properties methyl stub
Compound Azaindole
GTPS IC50 (nM) 2.5 nM Dissociation t½ (h) 46 ± 15 h
cLogP 4.8
cLogD 2.4
39 CRTh2: Can Residence Time Help ?
. For a purely antagonistic effect, prolonging Receptor Occupancy prolongs PD effect
. You will only really appreciate a PK-PD disconnection if Dissociation half-life >> PK terminal half-life
. For GPCR antagonism, you should count on extending the PD effect by half a half-life
. A “PK phase” of antagonism is needed to “set” the ligand in the receptor, regardless of mechanism
. We are pretty good at explaining what’s behind Structure-Activity Relationships (SAR) Structure-Residence Relationships (SRR) are in their infancy and/or qualitative
. To find Long Residence, you need to look for it
Efficacy
Residence Potency Time
40 Acknowledgements
Medicinal Chemistry Respiratory Therapeutic Area Computational and Structural Juan Antonio Alonso Mónica Bravo Chemistry and Biology Mª Antonia Buil Marta Calbet Sandra Almer Oscar Casado José Luís Gómez Yolanda Carranza Marcos Castillo Encarna Jiménez Sònia Espinosa Jordi Castro Martin Lehner Estrella Lozoya Paul Eastwood Isabel Pagan Cristina Esteve Pathology and Toxicology Manel Ferrer Ana Andréa Pilar Forns Mariona Aulí Elena Gómez Biological Reagents and Screening Cloti Hernández Jacob González Fani Alcaraz Neus Prats GalChimia Miriam Andrés Sara López Julia Díaz Development Marta Mir Teresa Domènech Cesc Carrera Imma Moreno Vicente García Nieves Crespo Sara Sevilla Rosa López Juan Pérez Andrés Bernat Vidal Adela Orellana Juan Pérez García Laura Vidal Sílvia Petit Gemma Roglan Pere Vilaseca Israel Ramos Enric Villanova Francisco Sánchez Carme Serra Intellectual Property ADME Integrative Pharmacology Houda Meliana Manel de Luca Clara Armengol Eulàlia Pinyol Peter Eichhorn Laia Benavent Laura Estrella Luis Boix Management Dolores Marin Elena Calama Paul Beswick Juan Navarro Amadeu Gavaldà Jordi Gràcia Montse Vives Beatríz Lerga Victor Matassa Miriam Zanuy Sonia Sánchez Monste Miralpeix
41