CRTh2: Can Residence Time Help ?
RSC-SCI Symposium on GPCR2 in Medicinal Chemistry
Allschwil, Basel 15-17 September 2014
Rick Roberts Almirall Barcelona, Spain Introduction
Paul Ehrlich, The Lancet (1913), 182, 445
“A substance will not work unless it is bound”
100 years on:
“What a substance does once it’ s bound may depend on how long it’s bound for”
2 The influence of Binding Kinetics Residence time / Dissociation half-life seconds minutes hours days level PD
PK
time
Mechanistic Surmountable Insurmountable Pharmacodynamic
Partial vs Full agonism Efficacy vs substrate concentration Duration of Action . M3 agonists . Lovastatin Candasartan . Ipratropium vs Aclidinium . A2A agonists Mechanism-based toxicity Kinetic Selectivity . Clozapine Haloperidol . M3 vs M2 antagonism . Celecoxib Aspirin
Potency has little influence over these behaviours
3 The CRTh2 Programme
4 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 . eosihiliinophil recruitment an d surv ilival . mucus secretion . Potential benefit in: . airway hyper-responsiveness . asthma . immunoglobulin E (IgE) production . allergic rhinitis . etc . atopic dermatitis.
5 CRTh2 – A Target of interest Indole acetic acids
HO O N N ?
F Oxagen-Eleventa AstraZeneca Actelion Novartis Merck Pulmagen-Teijin
OC-459 AZD-1981 Setipiprant R = CF3 QAW-039 MK-7246 ADC-3680 Phase III Phase II Phase III R = H QAV-680 Phase I Phase II Phase II/II Aryl acetic acids Phenoxyacetic acids
?
Boehringer Array Roche BI671800 ARRY-502 RG-7581 AstraZeneca Phase II Phase II Phase I AZD-5985 or AZD-8075 Phase I/I
. All compounds are acids
. Plenty of Competition Panmira Panmira Amgen . Plenty of Attrition AM461 AM211 AMG-853 Phase I Phase I Phase II . First compounds high dose and/or BID
6 Our internal programme We want to find a Once-a-day Oral low dose (≤ 10 mg) CRTh2 antagonist for mild-moderate asthma
. Therefore, we chose to deliberately look for slowly dissociating CRTh2 antagonists:
. To maintain receptor occupancy beyond level normal PK and extend duration of action PD
PK
. to reduce the pressure of finding a time carboxylic acid with desirable PK properties Pharmacodynamic
. to add the possibility of extra protection
due to insurmountability against PGD2 burst
Insurmountable
7 Assays
8 GTPSvsPGD2 binding
3 [ H]PGD2 M) (n 50 IC 2 D GG H]P 3 [
[35S]GTPS
35 Good correlation [ S]GTPSIC50 (nM) [35S]GTPS Assay favoured
9 GTPS vs ESC Isolated cell
CRTH2 (nM) 50 ange IC Whole cell hh l Shape C ii Eosinoph
Isolated Compounds show higher membranes potillltency in cellular assay
+ 0.1% BSA
35 Protein binding plays a role [ S]GTPSIC50 (nM)
10 GTPS vs ESC Human Whole Blood
CRTH2 ) More affinity for (nM
50 HSA > BSA e IC
Whole cell gg + 4% HSA pe Chan aa nophil Sh ii Eos
More affinity for membranes BSA > HSA
+ 0.1% BSA “Shift” assayypy simply reflects affinit y 35 of compounds for BSA and HSA. [ S]GTPSIC50 (nM)
ESC IC50 is “Real” Potency
11 GTPS vs DP1 Receptor Selectivity 10 µM
>50% DP1 DP1 inhibition at 10 µM ibition at hh r DP1 %In oo
PGD2 d Recept ii Prostano
CRTh2
35 [ S]GTPSIC50 (nM)
12 GTPS vs TP Receptor Selectivity
TXA2 10 µM tt
>50% TP TP inhibition at 10 µM hibition a nn tor TP %I tor TP pp
PGD2 ane Rece xx hrombo TT CRTh2
Compounds are [35S]GTPSIC (nM) CRTh2 selective 50
13 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 RdtReadout time 5000 Readout 4000 1 h 3000 Class ica l Surmountable 2000 Readout inhibition Ramatroban 1000
0 0.1 nM 10 nM 1 µM 100 µM
PGD2 Concentration 1000
800 Readout 15 min Classical 600 Insurmountable
eadout 400 inhibition RR
TM-30089 200 (CAY-10471) 0 0.1 nM 10 nM 1 µM 100 µM
PGD2 Concentration CAY-10471 0 → 5 µM
14 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 RdtReadout time 24 h 5000 Readout 4000 1 h 3000 Class ica l 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
eadout 4000
400 eadout RR RR 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
Surmoun ta bility ijis jus t a PGD2 Concentration PGD2 Concentration question of time CAY-10471 0 → 5 µM
15 When is the insurmountable surmountable ?
Classical Classical Surmountable inhibition Insurmountable inhibition Observational timescale
A B Competitive compounds B seems immobile in the with different kinetics timescale ex periment: Ins urmo untable
Observational timescale A and B reach equilibrium A B during the experiment. Surmountable behaviour timescale
PGD2 Release PGD2 Kinetics CRTh2 antagonist from Mast cells ~15 min Kinetics ~24 h
PGD burst PGD2 Metabolism 2 ~15 min timescale
Long resident CRTh2 antagonists will have an insurmountable behaviour against PGD2 burst
16 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 . Automated Filtration reading in 96-well format . Readings from 2 min to 28 h RdttiReadout times
100
80 % nn 60 Dissociation t1/2 < 1 min
40
Ramatroban 20 “Worst case” scenario for Mean Inhibitio dissociation half-life Surmountable 0 0 4 8 12 16 20 24 28 Time (h) . No rebinding possible. 100
80 . Phys iol ogi cal syst em may Dissociation t1/2 ~ 80 min 60 be less demanding
40 n Inhibition % aa 20
TM-30089 Me (CAY-10471) 0 0 4 8 12 16 20 24 28 Insurmountable Time (h)
17 Mechanisms of slow dissociation
18 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
Energy Energy
‡ (R·L)‡ (()RL)
(R·L)‡ slow Ea Ed
[R] + [L] [R] + [L] quick [RL] [RL] [RL*] Reaction coordinate Reaction coordinate
19 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 cy nn 80
However, the time to 60 “get on” is about 6 h. tor Occupa tor 40 pp
Before this time the 20 ligand is a partial % Rece antagonist 0 6 12 18 24 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
20 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 cy nn 80 Total RO% 60 RL*%
tor Occupa tor 40 pp RL% 20 % Rece 0 6 12 18 24 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
21 Which mechanism is it ?
. No radio -labelled ligands
. No Biacore
. Slow dissociating compounds get more potent with time Compound Dissociation half-life ) MM (n 50 IC
Changes in potency Compounds are not sufficient to active from t=0 estimate which mechanism is acting
“Real” potency may be better than in vitro potency
22 Which mechanism is it ? Induced fit model 100 k1 k3
[[]R] + [ []L] [[]RL] [RL*] ncy aa 80
k2 k4 Total RO% 60 RL*% 40 ptor Occup
ee RL%
2 h Pre-incubation at 10 × IC50 20
Dissociation t1/2 25 h ± 3.5 % Rec 0 6 121824 Time (h)
Majjyority [[]RL*] fast Dissociation
Residual [RL] fast Dissociation
MhiMechanism un known, biddfibut induced fit suspecte d
23 How long is long? Mechanistic or Pharmacodynamic? Necessary Receptor Occupancy for efficacy - Pharmacodynamic
TtTarget Class RtOReceptor Occupancy GPCR Antagonist 60-80% Agonist high efficacy 2-30% Agonist low efficacy 60-95% Ligand-gated ion channels Antagonist 65-95% Agonist 5-80% Transporters All 60-85% Enzyme Inhibitors 70-99% Grimwood and Hartig, Pharm. Therap. 122, 281, 2009 Expanded zone: 0 – 1half1 half-life Exponential Decay Exponential Decay 100 95% 100 90% y cy
cc 80% nn 80 Sometime here, a GPCR 80 70% antagonist will stop being 60% 60 effective after washout 60 r Occupan
or Occupa 40 tt 2 oo 40 20 3 4 20
Recep 5 Recept 0 0 0 2 4 6 8 10 0.0 0.2 0.4 0.6 0.8 1.0 Half-lives Half-lives For a GPCR antagonist, count on about half a half-life extra of PD effect
24 DoA Scenarios for full coverage over 24h Many CRTh2 antagonists in the clinic are twice daily
PK only effect: Some PK Little PK “classic” drug. PK-PD relationship Some residence time Really long residence time
PK PK K PK off Koff Cmax C PD effect max Plasma Residual levels activity at 24h IC IC50 50 IC50 t=0 24 h t=0 24 h
. Dissociation Half-life = . Dissociation Half-life . Dissociation Half-life not necessary ~ 24 h ~ 48 h
To turn a twice-daily compound into a once-daily compound, we want to add on a Dissociation half-life of ≥ 24h
25 Chemical Series
Structure-Activity Relationships (SAR) Struct ure-Kineti c RltiRelations hips ( SKR)
26 Are there slow-dissociating CRTh2 antagonists?
Ramatroban TM-30642 TM-30643 TM-30089 (CAY-10471)
Structure not disclosed Merck Amira Pulmagen MK-7246 PGD2 AM432 ADC-3680
Dissociation CdCompound PtPotency RfReference half-life*
Ramatroban pA2 36 nM 5 min Mol. Pharmacol. (2006), 69, 1441
TM-30642 pAp 2 20 nM 8 min --- / / ---
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
QAW-039 Kd 1 nM 12 min ERS 8 September 2014
QAV-680 Kd 15 nM 1min1 min ERS 8 September 2014 Published residence times are all “short”
27 Pyrazoles . Indole nucleus developed by Oxagen. Beginnings of slow dissociation observed
. First series of Pyrazoles gave active compounds, but no significant residence time (BMCL, 2013, 23, 3349)
. Second series of Reverse Pyrazoles gave a similar story (Eur J Med Chem, 2014, 71, 168)
Indole Pyrazoles Reverse Pyrazoles Oxagen X = CH X = N General Ph General
GTPSIC50 (nM) 14 7 4 4 – 100 35 32 – 450 Dissociation t½ (h) 1.3 0.2 1.9 0.02 – 0.7 0.2 0.04 – 0.7
SAR pretty good. No SKR advances observed in either core or tail sections
Pyrazoles series abandoned for general lack of Residence time
28 Pipas . Third series of Pyrazolopyrimidinones (Pipas) gave active compounds with long residence (BMCL Accepted for publication)
. Core SAR flat. Core SKR varied
Pyrazolopyrimidinones (Pipas)
* * * * * * *** * * *
Indole
Oxagen PiPa N-Me N-Bn diMe N-Me* N-CHF2*
GTPSIC50 (nM) 14 551 4 52 Dissociation t½ (h) 1.3 2.3 5.3 6.6 10 8 21
29 Pipas
. Tail SAR flat. Tail SKR varied
. Sulphone positioning ultimately affects SKR
O O S R
HO O
N
Ortho substitutionO Para substitution N H 1 2 R R GTPSIC50 Dissociation t½ R GTPSIC50 Dissociation t½ HH 5 nM 2.3 h Ph 170 nM n.d.
MeO H 4 nM 4.2 h Me 900 nM n.d.
H F 5 nM 33h3.3 h Bn 3 nM 090.9 h
MeO F 7 nM 23 h Ultimately Potent but no duration
. Pipa series was essentially impermeable.
. Not suitable for an oral programme.
. Series abandoned due to impermeability
30 Biaryl series . Fourth series of diverse biaryl compounds finally gave good activity and long residence time (BMCL Accepted for publication)
. Core SAR and SKR varied
N HO O O
CF3 A
Bicyclic Heterocycle
* * * * * * * * * * * * * * 6,5-ring 5,6-ring Amira Indazole Indazole Indole Indazole Indole Azaindole
GTPSIC50 (nM) 16 48 48 19 19 37 9 Dissoci ati on t½ (h) 2 141.4 121.2 404.0 161.6 252.5 232.3
31 Biaryl series – Indazole core
. 6 member ring SAR flat. SKR varied
1 2 R R GTPSIC50 Dissociation t½ HH 19 nM 1.6 h
Cl H 6 nM 3.2 h
F H 4 nM 5h
H Cl 16 nM 28h2.8 h
F Cl 15 nM 10.5 h
. Small substituents in R1 led to moderate increased in both potency and dissociation t1/2
. Substitution in R2 led to minimal changes in binding potency
GdGood star ting po int f or th e search of th e d esi red RidResidence time
32 Species differences
Percentage of identical residues among all ungapped positions between the pairs.
Human Guinea Pig Rat Mouse
Human 100% 73% 78% 80%
Guinea Pig 100% 69% 70%
Rat 100% 94%
Mouse 100%
How similar are these species ?
33 GTPS potencies human vs GP M) nn ( 50 Pig IC aa Guine
Similar Potencies
Human IC50 (nM)
34 GTP human vs GP Residence Time
Mechanistically useful
species t½ -life (h) Human 2 h ff
Guinea pig 29 h iation hal cc Pig Disso
Guinea Pharmacologically species t½ useful Human 23 h
Guinea pig 1.9 h Human Dissociation half-life (h)
35 PK-PD Disconnection Simulations
PK half-life 1 h
“PD” GPCR threshold A 1 h Dissociation half-life is barely noticeable in the PD Poor drug – short duration of action
A 12 h Dissociation half-life is definitely observable A short acting drug keeps working long beyond its expecttitations At t = 0,
[Ligand] = 10×IC50
PK half -life 12 h
“PD” GPCR threshold
A 1 h Dissociation half-life goes completely unnoticed Good PK means once-a-day dosing
A 12 h Dissociation half-life is barely noticeable A great drug is really efficacious over 24 h
At t = 0, Next day
[Ligand] = 10×IC50 Next dose For the greatest observable effect, Dissociation half-life >> PK half-life
36 For a recent article, see Drug Disc. Today (2013), 18, 697 PK-PD disconnection model 3 mg / kg dose Plasma Eos inophili a Timepoint diMe-Pyrrolopiridinone levels Inhibition guinea pig profile 1 h 1300 ng/ml 100% Eosinophilia IC50 3 ng/ml Dissociation t½ 20 h PK t½ 0.9 h 15 h undetectable 70%
%Inhibition of PK Eosinophilia K off 100 Short PK, Long residence time PK-PD disconnection 80
70 % PK Koff Residual 60 activity at 40 Residual 15h IC50 activity 20 at 24h 3 ngng//mlml IC50 0 t=0 24 h
37 PK-PD Disconnection in Guinea Pig Eosinophilia
Oxagen BiAry l PiPas
Eosinophilia IC50 (ng/ml) ~50 ~40 ~5
Gp Dissociation Half-life 1 h 15 h 20 h
Gp Pharmacokinetic Half-life 5 h 3 h 0.9 h
Oxagen PK-PD
2h 24h
Good PK OK PK Poor PK Short Dissociation Good Dissociation Good Dissociation No separation PK-PD Small separation PK-PD Large separation PK-PD
Long residence time translates to a PK-PD disconnection Remember: half a half-life
38 Molecular Determinants of Long Residence
39 Where is long Residence? Selected general reports or Structure Kinetic Relationships (SKRs)
. 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
Molecular size / weight ? Lipophilicity ? Charged state ? Rigidity ? Don’t know ?
Selected specific reports or Structure Kinetic Relationships (SKRs)
. Therapeutic Complement Inhibitors. J. Mol. Recognit. (2009), 22, 495 . Slow dissociation M3 antagonists. J. Med. Chem. (2011), 54, 6888 . CCR2 antagonists. J. Med. Chem. ((),2013), 56, 7706 . CDK8/CycC inhibitors. PNAS (2013), 110, 8081. . Adenosine A1 Receptor antagonists. J. Med. Chem. (2014), 57, 3213
40 Energetic concept of Residence Time Standard model Structure Kinetic Relationships (SKRs) k on hbllidhave been largely ignored: [R] + [L] [RL] Residence Time k unbound off bound Dissociation half-life SSlowlow oorr fastfast kineticskinetics Off-rate, k Energy SKR off (R·L)‡ The Dissociation rate is controlled by 2 factors:
Off rate (1) The potency (SAR)
koff (2) The Transition state energy
[R] + [L]
Potency
Ki [RL] (R·L)‡
PtPotency can be measure d If we can control the transition state energy, we can control the binding kinetics
41 Where is Long Residence ? Are more active compounds longer resident ?
CDK8/CycC inhibitors PNAS (2013), 110, 8081. Kd 5820 nM Kd 10 nM Diss t½ < 1.4 min Diss t½ 32 h
Energy (R·L)‡
~1 order of magnitude of “stickiness” [R] + [L] Longer resident?
More [RL] potent [RL]
Reaction coordinate
Clearly, Residence Time is linked to Potency
42 Where is long Residence?
Are more active compounds OOH Cl longer resident ? O Cl N ‡ H (R·L) OMe O O
Slower IC50 8 nM IC50 98 nM On/off Diss t½ 5i5 min Diss t½ 21 h Energy (R·L)‡ CRTh2 program data 4d4 orders of magn itdfitude of “stickiness”
[R] + [L] Longer resident?
More [RL] potent [RL]
Reaction coordinate
We are affecting the transition state more than the final binding position
43 Homing in on Residence Time in the Biaryl series
4-Azaindole Me-4-Azaindole
GTPSIC50 (nM) 14 16 Dissociation t½ (h) 131.3 21
A Magic Methyl for SKR ?
44 Homing in on Residence Time in the Biaryl series
IC50 5 nM IC50 8 nM Dissoc ia tion Dissoc ia tion half-life half-life 9 h 11 h
S HO O O IC50 4 nM N inactive HN Dissociation N half-life 2i25 min
Long Residence Time in the Biaryls comes from an H-Bond Acceptor in ortho
45 Amide Rotamers
O 50:50 by NMR N HO O CF3
IC50 14 nM Dissociation OMe half-life 2 h
N O
HO O CF3
OMe
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
46 H-Bond Acceptors – Steric requirements Do very subtle steric effects first determine Potency binding, then residence binding?
3 H H R H H H N N N O N N N N O N N N N
Me Fused Pyrazole Isoxazole Triazole Amide R3 Sterics Large Medium Small Minimum Minimum - Potency Low Medium High High High High Residence - Low Medium High High High
H H H H Steric Requirements for HBA N H O H N N R1 N H N Bn R3 ≤ H R3
Oxazole Pyrazole Pyrazole R1 > H R1 X R1 Sterics Minimum Small Large R2 R2 >H Potency Low Medium High RidResidence - - MdiMedium
47 Can we check the H-Bond Acceptor location?
O S O N HO O
N
F 2
Ab initio minimisation
Compound 1 23
GTPSIC50 20 nM 4 nM 6 nM Dissociation t½ 10 h 1.7 h 18 h
48 Can we achieve truly long residence? If we understand it, we can harness it.
Position-by-position analysis of good Benzyl to bump structural features for Residence Time up lipophilicity Minimum expression of 3º amide as potency and long residence H-bond acceptor Isoxazole as H-bond acceptor O Acetic acid N
HO O
Cl Chloro is N OK here N methyl stub
Azaindole Azaindole
GTPSIC50 (nM) 14 nM GTPSIC50 (nM) 2.5 nM Dissociation t½ (h) 1.5 h Dissociation t½ (h) 46 ± 15 h
cLogP 1.1 cLogP 4.8
cLogD -1.1 cLogD 2.4
Once-a-day purely from Residence Time ?
49 Lead Compound Profile In vitro and In vivo Physical Chemistry
human GTPSIC50 4 nM Solubility pH 1 and 7.4 > 1 mg/ml
ESC hWB IC50 3 nM LogD 1.3 Dissociation t½ (h) 22 h Caco (AB/BA) 24 / 6
gp GTPSIC50 (nM) 3 nM
Dissociation t½ (h) 14 h Pharmacokinetics Eosinophilia IC50 4 nM Rat Clearance 9ml/min/kg9 ml/min/kg Eosinophilia ID50 at 2 h 0.020 mg/kg Volume 0.5 L/kg PK-PD Disconnection Yes Terminal half-life 1.6 h Bioavailability 80% Safety Dog Clearance 2 ml/min/kg
hERG, Nav1.5, Cav1.2 > 10 µM Volume 0.9 L/kg Working heart Clean Terminal half-life 8 h Cytotox > 100 µM Bioavailability 51% GreenScreen Clean Pred. Human Clearance 2 ml/min/kg CYP3A4 20 µM Pred. Volume 1.4 L/kg Major metabolite Glucuronide Pred. Terminal half-life biphasic Acyl Glucuronide stability t½ > 4 h PK-PD dose simulation 2-6 mg QD
An oral, low dose (<10 mg), once-a-day CRTh2 Antagonist
50 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-Kinetic Relationships (SKR) are in their infancy and/or qualitative
. RidResidence Time can bdibe designe d
. To find Long Residence, you need to look for it
51 Acknowledgements
Medicinal Chemistry Respiratory Therapeutic Area Computational and Structural Juan Antonio Alonso Móni ca Bravo Chem is try an d Bio logy 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 Fani Alcaraz Cloti Hernández Jacob González 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 Villanov a Francisco Sánchez Carme Serra Intellectual Property ADME Integrative Pharmacology Houda Meliana Manel de Luca Clara Armengol Euuàlàli aPinyol Peter Eichhorn Laia Benavent Laura Estrella Luis Boix Management Dolores Marin Elena Calama Paul Beswick Juan Navarro Amadeu Gavaldà Jordi Gràcia MtMontse Vives BtíBeatríz Lerga Vict or MtMatassa Miriam Zanuy Sonia Sánchez Monste Miralpeix
52