Discovery PhysChem at UCB: AKAS and Beyond

Discovery PhysChem at UCB:AKAS and BeyondChristine Prosser UCB, Slough, UK PhysChem Forum 30th November 2005

Discovery PhysChem within UCB

Discovery PhysChem within UCBHighlight potential issues such as poor solubility, permeability and instability to allow accurate interpretation of biological results

Give greater confidence in-vitro and in-vivo data

Set up a cascade of assays to aid decision making in project teams

10 mM DMSO StockPPB(request)Log P(request)pKa(request)Discovery PhysChem (Slh)PhysChem (Brne + Cam)DMPK (Slh)QC CheckSnapSol(request)BioSol(all NCEs)AKAS(all NCEs)QuickSol(request)TSol(request)SolubilityGI PAMPA(all NCEs)BBB PAMPA(all NCEs)Caco-2(request)Permeability HT Log D7.4(all NCEs)Others

AKASAutomated Kinetic Aqueous SolubilityAll novel compounds pass through the assay (~100/week)Kinetic solubility measured across a pH rangeAllows estimate of pKaStability issues can be identified

AKAS - Experimental 10mM DMSO stock buffered at 4 pHs5% final DMSO concentrationSamples shaken for 90mins then filteredConcentration of sample in filtrate determined by UV analysis against calibration curve240-400nmmax / isosbestic pointSolubility of all compoundsmeasured in duplicate

AKAS Results (pKa from Solubility Profile) ChlorpromazineCalibrationSolubilityNo spectral changeBasic ionisation event between pH7.4 and pH9pKa ~9

AKAS Results (Spectral Changes)CalibrationSolubilityLabetololSpectral change upon ionisationIsobestic point used instead of maxAcidic ionisation event between pH5 and pH7.4Spectral change indicates ionisation event not apparent from solubility profile

AKAS Results (Stability Issues)CalibrationSolubilityCompound XCalibration shows no UV spectral changeSolubility curve at pH9 shows spectral changePossible stability issue at high pHND result returned

Kinetic vs. Thermodynamic Solubility?Kinetic solubility:Better suited to HTDoesnt take into account dissolution rateCo-solvent present Thermodynamic solubility:Expensive with respect to sample quantity and timeNot suited to HT

QuickSolPseudothermodynamic SolubilityStarts from solid materialSingle time point

QuickSol measured on requested compoundssolubility >150uM in AKAS at pH5 or pH7.4

Sheet1

ND0 - 30uM30 - 150uM150 - 350uM350 - 1500uM

PoorModestGood

Sheet2

Sheet3

QuickSol - ExperimentalBuffer added to solid sample0.5mg solid per pHmeasured in duplicate at 2 pHsSamples shaken for 90mins then filteredConcentration of sample in filtrate determined against calibration curveAnalysis by HPLC

QuickSol - ResultsComparison of AKAS and QuickSol results shows effect of DMSO on solubilityKetoconazole

AKAS and QuickSolAKAS best case scenario QuickSol worst case scenario

Combination of AKAS and QuickSol data used to highlight compounds where solubility may be a serious limitationinterpretation of biological results

BioSolSolubility under Biological Assay ConditionsPrimary assay data generated in enzyme or protein based assaysSolubility may be compromised by presence of proteins or incubation mediaCellular assays DMSO must be kept to minimum

All novel compounds pass through assayAssay measures solubility at relevant DMSO concentration in incubation media Same experiment carried out without protein to flag any potential protein binding issues (can then be investigated further by traditional DMPK methods)

BioSol - Experimental10mM DMSO stock diluted in D-PBS0.2% final DMSO concentration (sample concentration 20uM)10% bovine serum albumin10% glucoseMgCl2, CaCl2 (1M)Control plate without proteinShaken for 90mins then filteredFiltrate centrifuged through 10kDa cut off platesAnalysis by HPLC

BioSol - ResultsComparison of AKAS and QuickSol results shows no evidence of DMSO effect on solubilityBioSol result much lower than QuickSolComparison of BioSol with protein to BioSol blank suggests protein bindingWarfarin

Sheet1

AKAS pH7.4BioSol pH7.4QuickSol pH7.4

5% DMSO0.2% DMSO0% DMSO

>3509>1500

Sheet2

Sheet3

Sheet1

AKAS pH7.4BioSol pH7.4QuickSol pH7.4

5% DMSO0.2% DMSO0% DMSO

>3509>1500

BioSol with ProteinBioSol without Protein

920

Sheet2

Sheet3

SnapSolHT Single Point Solubility AssayDeveloped as an aid to fragment screening to ensure that compounds meet minimum solubility criteriaCan be used to screen a selection of compounds under conditions which mimic closely those of the screening experiment

Experimental10mM DMSO stock solutions are diluted in the buffer system used for the screening experimentbuffer typeDMSO concentrationShaken for 90 minutes then filteredAnalysis of the filtrate by UV plate reader at fixed wavelength against a calibration curve

PAMPAParallel Artificial Membrane Permeability AssayPermeability traditionally measured in in-vitro cell based assaysnot suitable for high throughputPAMPA measures passive diffusion through a lipid membraneno active transport mechanismsno efflux

All novel compounds pass through assays Used as a screening tool to select compounds for further permeability studies

GI PAMPA - ExperimentalPhosphatidyl choline artificial membrane dispensed onto a solid filter support10mM DMSO stocks diluted in buffer (100uM, 5% DMSO)PAMPA sandwich incubated at 20C for 4.5hrs, shaking at 750rpmAnalysis of carried out by UV plate readerPapp values returned

% membrane retention also calculated using mass balance equations

BBB PAMPABlood Brain Barrier PAMPAUsed to determine whether compounds are CNS+ or CNS-Important to UCB as we are targeting both CNS and non-CNS disease statesSame methodology as GI PAMPA but using porcine polar brain lipidL.Di, E.Kerns et al Eur. J. Med. Chem 38 (2003)223-232Assay validated against known CNS active and inactive drugs

PAMPA Results (GI vs. BBB)GI-PAMPA shows poor permeability.BBB-PAMPA shows good permeability. Consistent with literature, CNS+ drug.Oxazepam

Sheet1

GI-PAMPABBB-PAMPA

Pa (x 10-7 cm/sec)Pa (x 10-7 cm/sec)

1442

Sheet2

Sheet3

PAMPA Results (comparison with cell assays)GI PAMPA shows good permeabilityCaco-2 shows poor permeabilityEfflux mechanismsBoth GI and BBB PAMPA show poor permeability. But caffeine known to cross BBB - actively transported across BBBVerapamilCaffeine

Sheet1

GI-PAMPABBB-PAMPA

105

Sheet2

Sheet3

HT LogD7.4All novel compounds pass through the assay

Robotic based assay to measure partition between octanol and aqueous buffer

Used to rank compounds within a series and flag compounds that have LogD outside desired range.

HT LogD7.4 - Experimental10mM DMSO stock dissolved in 1:1 mixture of octanol and D-PBS (100uM, 5% DMSO)

Samples shaken for 90mins then centrifuged

Octanol and aqueous phases separated by liquid handling robot

Concentration of sample in each phase determined by HPLC

HT LogD7.4 Results (Std Drugs)* A. Avdeef, Absorption and Drug Development, John Wiley & Sons, 2003Metolazone

Chart4

1.8198829066

0.6928803707

-2.2840075136

-0.1941178432

2.6457860306

0.2307955712

0.3695428382

1.0027879199

1.6877581043

-1.0435921796

-1.4084284203

1.9025950645

3.1128283466

-0.116771378

1.2923565724

-0.0854014268

-1.3659293037

-0.562394577

-0.1854224777

0.815362793

1.6186694001

-1.2352835325

-0.129445491

-0.1764395771

-0.5351958222

0.2539002136

1.9480889863

0.2568290278

HT

Literature LogD7.4*

HT LogD7.4

Sheet1

Gold StdHT

2.081.82

0.860.69

-2.01-2.28

-0.07-0.19

3.452.65

0.340.23

0.620.37

1.381.00

2.161.69

-0.62-1.04

-0.24-1.41

2.171.90

3.833.11

-0.11-0.12

4.11.29

0.09-0.09

-0.46-1.37

-0.34-0.56

0-0.19

1.410.82

2.411.62

-0.53-1.24

0.08-0.13

0-0.18

0.03-0.54

0.630.25

2.511.95

1.120.26

Sheet1

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

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0

0

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HT

Literature LogD7.4

HT LogD7.4

Sheet2

Sheet3

HT LogD7.4 Results (UCB Compounds)

Chart1

2.6126602927

2.1676113684

2.1688297452

0.8350178805

2.3520908845

0.6960339246

1.9268224295

1.5014908354

2.4097306343

1.1627465488

2.4158841726

-0.484257408

3.7202496516

2.6828003

1.7391957306

-0.0806905398

2.4450535392

1.3032065937

1.552148192

1.6082944844

2.6616085211

1.7470927511

2.1395202607

2.4123231113

3.0268442178

2.7690362844

2.7793560771

2.8571586465

3.2830617889

2.3249187288

3.0468154139

1.8870153078

2.8135958722

2.8435219985

1.8809882344

2.718893941

3.93

4.39

1.5462879398

4.2849237561

0.2040882439

2.8367812416

0.7555815366

2.3227993518

Shake Flask LogD 7.4

HT LogD 7.4

SampleList

Compound IDBATCH_IDsLogDProjectLogD (1)LogD - LogD (1)LogD (2)LogD - LogD (2)

1CT531833-0010016374/30/13.19AKT2.580.612.640.55

2CT535956-0010016337/69/12.94PI-32.440.502.390.55

3CT537066-0010016083/102/13.05P38-SLH2.640.412.680.37

4CT537395-0010016653/29/23.56PI3 Kinase3.050.513.000.56

5CT537434-0010016640/17/32.82AKT2.160.662.180.64

6CT537702-0010016653/43/13.35PI3 Kinase2.820.532.720.63

7CT538349-0010016595/35/12.74AKT2.140.602.200.54

8CT538842-1010016894/22/11.53AKT0.780.750.890.64

9CT539633-0010016760/19/12.1P38-SLH1.760.341.740.36

10CT539650-0010016410/88/12.53P38-SLH2.180.352.100.43

11CT539656-0010016820/14/13.35PI3 Kinase2.760.592.800.55

12CT539662-0010016849/7/23.32PI3 Kinase2.880.442.830.49

13CT539795-0010016558/82/12.82AKT2.330.492.370.45

14CT539810-0010016674/77/14.14PI3 Kinase3.360.783.210.93

15CT539851-1010016642/92/12.84PI3 Kinase2.240.602.410.43

16CT540010-0010016737/34/14.62PI3 Kinase3.011.613.091.53

17CT540111-0010016842/51/12.24PI3 Kinase1.860.381.910.33

18CT540324-0010016849/26/14.2PI3 Kinase2.871.332.761.44pKa 6 (0% DMSO)

19CT540469-0010016894/20/11.17AKT0.690.480.700.47

20CT540906-0010016737/46/24.76PI3 Kinase2.702.062.991.77

21CT540920-0010016960/27/12.1PI3 Kinase1.860.241.910.19

22CT540996-1010016856/34/42.41AKT1.880.531.970.44

23CT541048-11310016558/96/11.84AKT1.470.371.530.31

24CT541054-0010016742/21/13.02PI3 Kinase2.540.482.900.12

25CT541145-0010017504/10/13.77PI3 Kinase3.240.533.93-0.16

26CT541185-0010017081/6/23.02AKT2.390.632.430.59

27CT541588-0010017081/18/11.56AKT1.160.401.170.39

28CT542563-0010017081/30/13.01AKT2.390.622.440.57

29CT542564-0010016963/103/15.5PI3 Kinase4.391.110.000.00

30CT542737-0010016929/70/12.12PI3 Kinase1.520.601.580.54

31CT543986-1010017121/49/1-0.8AKT-0.47-0.33-0.50-0.30

32CT544072-0010017081/68/11.37AKT4.02-2.653.42-2.05pKa between 5 + 7.4 (5% DMSO)

33CT544133-0010017192/40/23.29AKT2.510.782.850.44

34CT544171-0010017081/94/12.3AKT1.730.571.750.55

35CT545311-0010017237/88/14.29PI3 Kinase4.33-0.044.240.05

36CT545728-0010017320/29/13.48PI3 Kinase0.163.320.253.23

37CT546467-0010017204/74/12.75AKT0.142.61-0.303.05

38CT546585-0010017320/87/13.2PI3 Kinase2.810.392.860.34

39CT546599-0010017249/84/10.91PI3 Kinase0.740.170.770.14

40CT546692-0010017544/1/12.72PI3 Kinase2.300.422.340.38

41CT546947-0010017569/8/12.98AKT2.400.582.490.49

42CT547006-0011017548/26/21.7AKT1.210.491.400.30

43CT547139-0010017569/26/11.91AKT1.550.361.560.35

44CT537710-0010016640/20/22.04AKT1.640.401.580.46

45Atenolol-2.01-1.74-0.27-1.92-0.09

46Sulfasalazine0.080.21-0.130.19-0.11

47Chlorpromazine3.452.800.652.311.14

48DMSO0.000.000.000.00

Results (1)

OctanolAqueous

Compound IDRTnmWellInj VolAreaWellInj VolAreaOctanolAqueousLogD

1CT531833-000.832280A112254.6A28026.51018426.52.5846725167

2CT535956-000.807254B12514.3B28075.22057275.22.4370586751

3CT537066-001254C12576.6C28052.42306452.42.6436033424

4CT537395-000.837280D12492.8D28017.51971217.53.0516926418

5CT537434-000.901254E12408.9E280114.216356114.22.156010998

6CT537702-000.903280F12423.5F28025.71694025.72.8189802827

7CT538349-000.722280G12306.1G28089.31224489.32.137071862

8CT538842-100.832254H12645.4H22107.1645.4107.10.7800394902

9CT539633-000.884254A32653.6A480458.826144458.81.7557486256

10CT539650-000.934254B32625.7B480164.425028164.42.1825243331

11CT539656-000.858280C32354.9C48024.61419624.62.7612308836

12CT539662-000.972280D32589.8D4803123592312.8814030657

13CT539795-000.88254E32578.2E480108.223128108.22.3299108179

14CT539810-001.134280F32260.5F4804.6104204.63.3551098873

15CT539851-100.794280G32468.5G480106.718740106.72.2446051671

16CT540010-001.128280H32349.1H48013.71396413.73.008289273

17CT540111-000.91280A52142.5A68078.2570078.21.8626681026

18CT540324-000.975280B52385.4B68020.71541620.72.8720013562

19CT540469-000.812280C52438.8C6289.4438.889.40.6909291001

20CT540906-001.133280D52375.7D68029.91502829.92.7012299981

21CT540920-000.814254E52336.7E680187.413468187.41.8565335212

22CT540996-100.868280F52413.5F680217.316540217.31.8814757789

23CT541048-1130.819280G52330.4G680447.713216447.71.4701129356

24CT541054-001.061280H52413.8H68048.21655248.22.5358034394

25CT541145-001.061254A7286.5A8802346023.2380461031

26CT541185-000.893280B72394.8B8806415792642.3922571613

27CT541588-000.785254C72318.4C880881.812736881.81.1596629559

28CT542563-000.892280D72359.9D88058.51439658.52.3910859719

29CT542564-001.266280E72492.6E8800.8197040.84.3914644118

30CT542737-000.854280F72467.9F880570.918716570.91.5156529935

31CT543986-100.691280G7233.3G8803953.313323953.3-0.4724555476

32CT544072-000.858254H721168.2H8804.5467284.54.0163646795

33CT544133-000.853280A92304.6A108037.31218437.32.5140810585

34CT544171-000.857254B92376.9B1080282.315076282.31.7275752506

35CT545311-001.171280C92322.4C10800.6128960.64.3323037741

36CT545728-000.931254D922.7D108074.910874.90.1589419378

37CT546467-000.748280E92191.4E10805559.276565559.20.1389896261

38CT546585-000.969254F92427F108026.51708026.52.8092419924

39CT546599-000.826280G9291.9G1080667.43676667.40.7409893004

40CT546692-000.931254H92231.9H108046.3927646.32.3017797489

41CT546947-000.878280A112520.9A128083.32083683.32.3981693474

42CT547006-000.888280B112195.8B1280487.57832487.51.2058980588

43CT547139-000.899280C112439.2C1280499.117568499.11.5465347529

44CT537710-000.874254D112454.8D1280421.218192421.21.6353920848

45Atenolol0.676280E1120.7E12801545.9281545.9-1.7420233658

46Sulfasalazine0.875254F11231.3F1280767.21252767.20.2126957347

47Chlorpromazine0.958280G11264.3G12804.125724.12.7974871075

Results (2)

OctanolAqueous

Compound IDRTnmWellInj VolAreaWellInj VolAreaOctanolAqueousLogD

1CT531833-000.831280A1110879.8A28016.17038.416.12.6406480686

2CT535956-000.806254B1101318.2B28043.210545.643.22.3875875475

3CT537066-000.988254C1102104.1C28035.216832.835.22.6796136999

4CT537395-000.834280D1101657.6D28013.213260.813.23.0019957939

5CT537434-000.894254E1101595.8E28084.512766.484.52.1792117388

6CT537702-000.898280F1101453.3F28022.211626.422.22.7190922861

7CT538349-000.721254G110710.2G28035.85681.635.82.2005876283

8CT538842-100.83280H1101359.1H2801400.710872.81400.70.8899962708

9CT539633-000.882280A3102444.2A480357.119553.6357.11.7384368766

10CT539650-000.928280B3102218.3B480142.117746.4142.12.0965161883

11CT539656-000.855280C3101270.3C48016.210162.416.22.7974812705

12CT539662-000.963280D3102144D48025.21715225.22.8329142272

13CT539795-000.876280E310784.2E48026.56273.626.52.374270951

14CT539810-001.135254F32101.6F4802.540642.53.2110136906

15CT539851-100.794280G3101798.7G48056.614389.656.62.4052322905

16CT540010-001.129254H32979.8H48032.23919232.23.0853415548

17CT540111-000.905280A510583A68057.2466457.21.911362513

18CT540324-000.96280B5101806.9B68025.414455.225.42.7551903883

19CT540469-000.809280C510961.1C6801530.17688.81530.10.7011387491

20CT540906-001.134280D52283.1D68011.71132411.72.9858139989

21CT540920-000.81280E510499.7E68049.73997.649.71.9054429477

22CT540996-100.863280F5101640.2F680139.913121.6139.91.9721690801

23CT541048-1130.815280G5101400.6G680328.511204.8328.51.5328687351

24CT541054-001.012254H510588.5H6805.947085.92.9019844425

25CT541145-001.019280A710423.5A8800.433880.43.9278834103

26CT541185-000.886280B7101678.1B88050.213424.850.22.4272041072

27CT541588-000.781254C7101342.1C880732.910736.8732.91.1658301417

28CT542563-000.887280D7101417.21D88041.111337.6841.12.4406823732

29CT542564-001.282254E7101004.6E88008036.800

30CT542737-000.854280F7101830.9F88038814647.23881.5769228861

31CT543986-100.69280G710153.5G8803848.212283848.2-0.4960592685

32CT544072-000.858280H72929.4H8801437176143.4241346238

33CT544133-000.855254A92565.5A1010039.82827539.82.8515195415

34CT544171-000.855254B92377.9B1080268.315116268.31.7508162106

35CT545311-001.112254C92172.8C101000.586400.54.2375437381

36CT545728-000.934280D923D108067.612067.60.2492345501

37CT546467-000.75254E9285.7E1010855.7428.5855.7-0.3003707057

38CT546585-000.971280F92250.6F108013.71002413.72.8643204908

39CT546599-000.828280G9273.9G1080501.82956501.80.7701737727

40CT546692-000.933280H92202.5H108036.7810036.72.3438189546

41CT546947-000.879280A112433.8A128055.91735255.92.4919377311

42CT547006-000.889280B112308.7B1280491123484911.4005151287

43CT547139-000.891280C112384.6C1280425.915384425.91.5577616312

44CT537710-000.875280D112209.3D1280219.68372219.61.5811968839

45Atenolol0.659280E1120.5E12801666201666-1.9206450014

46Sulfasalazine0.877254F11231F1280800.21240800.20.1902231381

47Chlorpromazine0.962280G11247.3G12809.218929.22.3131333047

By Project

Compound IDLogDProjectLogD (1)LogD - LogD (1)LogD (2)LogD - LogD (2)Ave

CT531833-003.19AKT2.580.612.640.552.61

CT537434-002.82AKT2.160.662.180.642.17

CT538349-002.74AKT2.140.602.200.542.17

CT538842-101.53AKT0.780.750.890.640.84

CT539795-002.82AKT2.330.492.370.452.35

CT540469-001.17AKT0.690.480.700.470.70

CT540996-102.41AKT1.880.531.970.441.93

CT541048-1131.84AKT1.470.371.530.311.50

CT541185-003.02AKT2.390.632.430.592.41

CT541588-001.56AKT1.160.401.170.391.16

CT542563-003.01AKT2.390.622.440.572.42

CT543986-10-0.8AKT-0.47-0.33-0.50-0.30-0.48

CT544072-001.37AKT4.02-2.653.42-2.053.72

CT544133-003.29AKT2.510.782.850.442.68

CT544171-002.3AKT1.730.571.750.551.74

CT546467-002.75AKT0.142.61-0.303.05-0.08

CT546947-002.98AKT2.400.582.490.492.45

CT547006-001.7AKT1.210.491.400.301.30

CT547139-001.91AKT1.550.361.560.351.55

CT537710-002.04AKT1.640.401.580.461.61

CT537066-003.05P38-SLH2.640.412.680.372.66

CT539633-002.1P38-SLH1.760.341.740.361.75

CT539650-002.53P38-SLH2.180.352.100.432.14

CT535956-002.94PI-32.440.502.390.552.41

CT537395-003.56PI3 Kinase3.050.513.000.563.03

CT537702-003.35PI3 Kinase2.820.532.720.632.77

CT539656-003.35PI3 Kinase2.760.592.800.552.78

CT539662-003.32PI3 Kinase2.880.442.830.492.86

CT539810-004.14PI3 Kinase3.360.783.210.933.28

CT539851-102.84PI3 Kinase2.240.602.410.432.32

CT540010-004.62PI3 Kinase3.011.613.091.533.05

CT540111-002.24PI3 Kinase1.860.381.910.331.89

CT540324-004.2PI3 Kinase2.871.332.761.442.81

CT540906-004.76PI3 Kinase2.702.062.991.772.84

CT540920-002.1PI3 Kinase1.860.241.910.191.88

CT541054-003.02PI3 Kinase2.540.482.900.122.72

CT541145-003.77PI3 Kinase3.240.533.93-0.163.93

CT542564-005.5PI3 Kinase4.391.114.39

CT542737-002.12PI3 Kinase1.520.601.580.541.55

CT545311-004.29PI3 Kinase4.33-0.044.240.054.28

CT545728-003.48PI3 Kinase0.163.320.253.230.20

CT546585-003.2PI3 Kinase2.810.392.860.342.84

CT546599-000.91PI3 Kinase0.740.170.770.140.76

CT546692-002.72PI3 Kinase2.300.422.340.382.32

By Project

Rep 1

Rep 2

10 mM DMSO StockPPB(request)Log P(request)pKa(request)Discovery PhysChem (Slh)PhysChem (Brne + Cam)DMPK (Slh)QC CheckSnapSol(request)BioSol(all NCEs)AKAS(all NCEs)QuickSol(request)TSol(request)SolubilityGI PAMPA(all NCEs)BBB PAMPA(all NCEs)Caco-2(request)Permeability HT Log D7.4(all NCEs)BIO-PAMPA(all NCEs) HT pKa(all NCEs)Others

Sheet1

AKASQuickSolBioSolPAMPABBB PAMPA

pH3 (uM)pH5 (uM)pH7.4 (uM)pH9 (uM)pH7.4 (uM)pH7.4 (uM)PPB?Pa (10-7 cm/sec)%RPa (10-7 cm/sec)LogD7.4

AlprazolamND>350>350>35082037301.79

Alprenolol>350>350>350>350ND20122260.66

Amiloride>350>350>350>350>15002000-1.24

Astemizole>350>350681302012563193.31

Atenolol>350>350>350>350>15002070-2.03

Caffeine>350>350>350>350>150020105-0.21

Chlorpromazine>350>350>35052>150094122.51

Corticosterone>350>350>350>350526204ND1.74

Desipramine>350>350>350>350>150020222280.93

Dexamethasone>350>350>350>350275200161.66

DiazepamND>350>350>35017214Yes17516252.72

Diltiazem>350>350>350>350>150020223381.66

Enoxacin>350>350>350>350>150020118-1.31

Famotidine>350>350>350244>15001900-1.08

Furosemide283>350>350>350>150000-1.44

Hydrocortisone>350>350>350>350123720001.29

Imipramine>350>350>350>350>15002011640171.84

Isoxicam2>350>350>350>150014Yes00-0.54

Ketoconozole>350>350252236711Yes633283.57

Ketoprofen>350>350>350>350>15007Yes01-0.16

Labetolol>350>350>350>350>15000780.71

Metolazone>350>350>350>350109031.29

Naproxen164>350>350>350>15008Yes00-0.15

Nifedipine>350>350>350>35020161331213ND

Norfloxacin>350>350>350>350>15002000-1.30

Ofloxacin>350>350>350>350>15002000-0.56

Oxazepam>350>350>350>3507613Yes14421.88

Piroxicam>350>350>350>350>150014Yes106-0.33

Promazine>350>350>350>350>150019319121.89

Propranolol>350>350>350>350>150020109320.75

Quinidine>350>350>350>350>1500209281.60

Ranitidine>350>350>350>350>15001800-1.21

Sulfasalazine144>350>350>350>15009Yes00-0.04

Sulpiride>350>350>350>350>15001700-1.14

Tenoxicam>350>350>350>350>15001662ND

Terfenadine>350>350204001700ND

Testosterone>350>350>350>350199201213.90

Theophylline>350>350>350>350>15002000-0.17

Timolol>350>350>350>350>150020013-0.52

Trimethoprim>350>350>350>350115920010.30

Verapamil>350>350>350>350>15002018813251.98

Warfarin>350>350>350>350>15009Yes210.20

Sheet2

AKASQsolBioSolPAMPABBB PAMPA

pH3 (uM)pH5 (uM)pH7.4 (uM)pH9 (uM)pH7.4 (uM)pH7.4 (>20uM)PPBPa (10-7 cm/sec)%RPa (10-7 cm/sec)

>350>3506813Yes6176327

Sheet3

pH3

pH5

pH7.4

pH9

Sheet3

1.77692.18622.19692.1483

1.76042.15312.16372.1173

1.75262.13012.14242.0962

1.75112.1182.13042.0837

1.75212.10322.11582.0683

1.75872.07352.08862.0396

1.77062.04782.06272.0108

1.78672.02062.03471.9843

1.79431.99962.01491.964

1.80191.9681.98351.9325

1.811.91481.92981.8813

1.81481.85421.86851.8211

1.81761.77441.78741.74

1.81631.71341.72541.68

1.81121.64591.65811.6149

1.79791.54451.5551.5155

1.78451.45451.4631.4258

1.76981.36911.37541.3379

1.75381.27971.28481.2515

1.73841.19941.20191.1696

1.72391.13661.13791.1061

1.71281.09451.0951.0639

1.70151.05991.061.03

1.68761.02551.02540.9969

1.66860.98760.98620.9586

1.620.94810.9440.9147

1.59960.92460.92050.8919

1.57380.89790.89410.865

1.54980.87640.87260.845

1.52060.85640.85270.8251

1.48440.83260.82930.8022

1.44630.80810.80630.7803

1.40540.78490.78190.7576

1.3710.76580.76260.7384

1.3380.74960.74670.7233

1.29210.72810.72580.7032

1.2480.70830.70590.685

1.20140.68780.68530.6639

1.16160.67030.66830.647

1.1220.65350.6520.6311

1.08340.63750.6360.6158

1.04130.61980.61850.599

1.00150.60250.60130.5823

0.96940.58890.58790.5692

0.93680.57410.57290.5547

0.90290.55790.55730.5399

0.86820.5410.54020.5232

0.83480.52380.52340.5069

0.8150.51370.5130.4968

0.78310.49630.49610.4806

0.75010.47770.47780.4632

0.72350.46130.46220.4481

0.6930.44290.44280.4301

0.66940.42790.42920.4164

0.6340.40510.40680.3953

0.60590.38680.38730.377

0.57770.36910.36960.3601

0.54850.35030.35080.3412

0.52090.3330.33310.3237

0.49660.31780.3180.3093

0.4720.30260.30290.2949

0.44080.28330.28360.2771

0.41620.26780.26780.2619

0.38750.24850.24880.2425

0.36150.22990.23010.2248

0.3340.20910.20940.205

0.30730.18760.18780.1843

0.28060.16520.16540.1629

0.25850.14580.1460.144

0.23520.1250.12480.1232

0.21470.10670.10620.1051

0.19630.09030.08960.089

0.17920.07510.07440.0741

0.16620.0640.0630.0632

0.14930.04980.04890.0495

0.13770.04070.03930.0399

0.1280.03340.0320.0328

0.11970.02770.02620.026

0.11110.0220.02040.0205

0.10430.01780.01610.0161

0.10010.01560.01390.0137

0.09480.01280.01150.0115

0.09060.01120.00940.0096

0.08670.00950.00780.0075

0.08410.00860.00680.0076

0.08130.00770.00610.0069

0.07920.00730.00560.0061

0.07690.00650.00510.0049

0.07510.00620.00490.0046

0.07280.00610.00440.0043

0.07080.00550.00410.004

0.06890.00510.00280.0034

0.06710.00490.00360.0034

0.06510.00460.00320.0028

0.06360.00430.00310.0027

0.06180.00420.00310.0025

0.05980.0040.0030.0024

0.05830.0040.00290.0022

0.05640.00380.00290.0023

0.05450.00370.00190.0024

0.05260.00340.00170.0024

0.05080.00350.00180.0023

0.04880.00350.0020.0025

0.04670.00310.00170.0025

0.04440.00320.00180.0023

0.04240.0030.00150.0022

0.04040.00280.00230.0024

0.03890.00260.00220.0023

0.03710.00270.00230.0024

0.03480.00290.00240.002

0.03320.00260.00120.002

0.03020.00260.00140.0019

0.02890.00250.00120.0017

0.02660.00240.00120.0017

0.02490.00250.00140.0016

0.02320.00260.00140.0018

0.02140.00230.00220.0015

0.01970.00250.00230.0017

0.01810.00260.00240.0017

0.01680.00240.00230.0017

0.01540.00230.00210.0028

0.0140.00240.00220.0026

0.01280.00290.00250.0027

0.01170.00260.00240.0027

0.01090.00230.00230.0028

0.00970.00220.00220.0029

0.00910.00270.00250.0029

0.0080.00280.00290.0019

0.00750.00270.00260.0022

0.00660.00290.00170.0025

0.0060.00240.00230.003

0.00540.00260.00240.0028

0.00480.00270.00280.0029

0.00430.00270.00260.0027

0.00380.00290.00260.0029

0.00310.00250.00220.0027

0.00310.00250.00250.0027

0.00250.00250.00220.0025

0.00220.00250.00240.0025

0.00210.00240.00250.0026

0.00120.0020.00210.0021

0.00080.00190.00170.0021

0.00090.00190.00190.0021

0.00080.00210.00190.0021

0.00060.0020.0020.0024

0.00060.00190.00220.0021

0.00040.00190.0020.0022

0.00030.0020.00210.0023

0.00030.00210.00180.0022

0.00030.00190.0020.002

0.00010.00190.0020.0019

0.00010.00190.00190.0021

00.00190.0020.0021

00.00180.00180.0019

00.00190.00180.0019

0.00010.00190.00190.002

0.00020.00190.0020.0021

00.00190.00190.002

-0.00010.00190.0020.0022

00.00190.00180.002

0.00010.00190.00210.0021

Solubility Data: Alprazolam

3.70753.31933.17233.1699

3.75223.21123.05333.0645

3.79243.08612.93132.9545

3.81133.00362.84962.8573

3.74472.9082.75772.7732

3.69132.76772.63112.6359

3.562.67462.53942.5399

3.4322.61032.4752.4837

3.33932.5742.44562.4498

3.20832.53462.41012.4129

3.01472.47722.36162.3655

2.82262.43172.31592.319

2.62152.38072.27112.2707

2.49762.34182.23452.2338

2.37322.2922.18852.1871

2.22942.21592.11592.1192

2.11582.13962.04442.0458

2.01982.05631.96411.9661

1.92311.95831.87071.8749

1.83621.84681.76271.7679

1.76261.74761.66631.6709

1.71081.66581.58941.5932

1.6641.5871.5151.5194

1.61521.49681.42861.4349

1.56251.39641.33091.3368

1.50771.28061.2171.2161

1.47391.20661.14631.1459

1.44091.12841.07161.0704

1.4171.06521.01061.0102

1.39951.01130.95930.9588

1.38180.94660.89720.8969

1.37020.89070.84370.8439

1.36310.83670.79260.793

1.3610.79780.75280.7539

1.36230.76840.72470.7249

1.36640.72860.68670.6884

1.37370.69560.65440.6559

1.38490.66390.62330.6238

1.3960.63830.59850.5984

1.40690.61570.57660.5763

1.41810.59270.55430.5541

1.42780.56940.53170.5317

1.43570.54580.50880.5088

1.43840.5280.49150.4912

1.43760.50820.47220.4719

1.43040.48580.45090.4513

1.41860.46290.42870.4289

1.4010.43810.40620.4064

1.38860.42470.39290.3931

1.36010.39930.36940.3698

1.33140.37690.34820.3489

1.30370.35790.32910.3308

1.27250.33950.31140.3127

1.24560.32610.2990.3005

1.20960.31080.28490.2861

1.1770.29980.27420.2743

1.14790.29320.26810.2677

1.11650.28790.26330.2627

1.08660.28560.26120.2601

1.06130.28550.26130.2601

1.03280.28720.26330.262

1.00270.29090.26730.2657

0.97590.29560.2720.2711

0.94810.30110.27790.2767

0.92120.3070.28480.2825

0.89220.31430.29130.2888

0.86330.32040.29760.2953

0.83220.3260.30360.301

0.80580.33010.30810.3054

0.77530.33280.31220.3096

0.74630.33520.31490.3123

0.71590.33690.3170.3143

0.6850.33770.31770.3151

0.65680.3370.31710.3147

0.61620.33390.31510.3126

0.58820.33060.31220.3099

0.55880.32530.30770.3057

0.52910.31870.30170.2988

0.49530.30880.29280.2903

0.46750.29880.28320.2808

0.44720.28950.27490.2725

0.41810.27380.26050.2586

0.39590.25940.24670.2454

0.37270.2420.23020.2288

0.35710.22860.21740.2172

0.34110.21250.20240.2025

0.32780.1970.18750.1876

0.31430.17950.17090.1702

0.30350.16370.15580.1554

0.29370.14750.13990.1395

0.28580.13270.12570.1254

0.2790.11690.10960.1105

0.27390.10470.09890.0986

0.26960.09090.08510.085

0.26740.08150.07630.0759

0.26580.07130.06650.0658

0.26540.06070.05610.0556

0.26550.05440.050.0494

0.2670.04630.04210.0413

0.26880.040.03510.0355

0.27180.03450.02970.0302

0.2750.02970.02490.0254

0.27880.02580.02120.0213

0.28250.02220.01780.0182

0.28720.01910.01470.0151

0.29070.0170.01240.0129

0.29540.01480.01110.011

0.29920.01340.00990.0095

0.30390.01230.00850.0082

0.30910.01140.00750.0068

0.31370.01040.00540.0059

0.31780.00950.00470.0049

0.32210.00980.0050.0049

0.32630.00860.00360.0036

0.32930.00830.00330.0033

0.33190.00810.00290.003

0.33460.00760.00340.0027

0.33720.00760.00340.0025

0.33850.00770.00350.0023

0.340.00740.00310.0022

0.34060.00710.00280.0032

0.34010.00720.00310.0029

0.34050.00770.00320.0029

0.34040.00730.00290.0028

0.33960.00710.00290.0029

0.33730.0070.00270.003

0.33530.00740.0030.003

0.33250.00730.00330.002

0.32940.00730.00320.0022

0.32520.00810.00310.0029

0.32060.00680.00290.003

0.31590.0070.00290.003

0.31110.00690.00320.0028

0.30450.00670.0030.0026

0.29850.00680.0030.0029

0.29150.00650.00260.0026

0.28480.00660.00290.0028

0.2760.00620.00260.0024

0.26780.00630.0030.0025

0.25930.00590.00290.0026

0.24880.00520.00240.002

0.23910.00510.00210.0021

0.22920.00510.00230.002

0.2180.00470.00220.0019

0.20710.00460.00230.0023

0.19610.00450.00250.002

0.1860.00430.00230.002

0.17620.00420.00230.0021

0.16480.00410.00210.002

0.15470.00380.00220.0019

0.14360.00370.00240.0019

0.13340.00350.0020.0019

0.1250.00340.00240.0019

0.11530.00310.00220.0018

0.10870.0030.00220.0017

0.09730.00290.00220.0018

0.09010.00290.00220.0019

0.08380.00270.00220.0019

0.07630.00260.00220.0021

0.06940.00240.00210.0019

0.06170.00240.00230.0018

Solubility Data: Diazepam

ConclusionsIn our opinion, data should be used as a packageno single piece of data should make or break a project

Provide information to allow an informed interpretation of results

AcknowledgementsPhysChem, Slough, UKRichard TaylorBarbara MasonEmily Freeman

PhysChem, Cambridge, UKJohn CooperBenedicte FauDave Sherwood

PhysChem, Braine, BelgiumLuc QuereLiliane EllensGeraldine Longfils

DMPK, Slough, UKTed PartonLloyd KingHanna HailuMark BakerSarah BartlettSimon CarterJudith van Asperen

Documents

Discovery PhysChem at UCB: AKAS and Beyond