INTRODUCTION

Seventh Wave was established in August 2003, and during its six years it has quickly grown from five employees to over fifty.

Deqing Xiao, Annie Walker, Lucy Hestco, Roger Melton, Andrew Vick Pharmacokinetics, Dynamics & Metabolism, Seventh Wave Laboratories LLC, 743 Spirit 40 Park Drive, Suite 209,Chesterfield, MO 63005

The BIOANALYTICAL LABORATORY

The Bioanalytical Laboratory (BA Lab) of Seven Wave was founded in 2006. It works closely with the In-Life and PK groups as a part of the Pharmacokinetics, Dynamics and Metabolism (PDM) Department. Located approximately 30 miles west of downtown St. Louis, Missouri, this new BA service lab is focused on the drug discovery needs of our clients with an emphasis on responsiveness, speed, integrity, and teamwork.

BIOANALYTICAL CAPABILITIES

LC-MS/MS method development and validation Bioanalytical support for discovery studies Rapid analysis of samples from multiple matrices Plasma and whole blood analysis Extensive options in tissue analysis Urine and/or feces analysis Determination of PK parameters and PK/PD relationships Studies requested by clients

In Vitro CAPABILITIES

Metabolic stability in hepatocytes and microsomes Intrinsic clearance Plasma protein binding Suspension homogeneity Test article stability as formulated Ligand binding assays

SUPPORTED PHARMACOLOGY MODELS

ACKNOWLEDGEMENT

We thank E. Schmidt and J. DeBold for their kind contribution to this poster.

CONTACTS

For additional information please contact:

Andrew Vick: avick@7thwavelabs.com

Roger Melton: rmelton@7thwavelabs.com

Deqing Xiao: 636-519-4873 (O), dxiao@7thwavelabs.com

Cardiovascular

• ApoE -/- transgenic mice

• LDL receptor deficient mice

Hepatic

• Hepatic injury and fibrosis (carbon tetrachloride-induced)

Oncology

• A/J mouse lung tumor studies

• NNK-induced lung tumors

Metabolic/Diabetes

• db/db mice

• ob/ob mice

• ZDF rats

• Chemically induced diabetes (STZ)

Respiratory

• Rhinitis

• Bronchitis

Additional models are under evaluation.

BA LAB FACILITY

The BA lab is equipped with state-of-the-art technology that is constantly updated to afford our clients the most modern research and analytical tools. Our LC-MS/MS/UV instrumentation is built around Sciex API 4000 QqQ and 4000 Q Trap MS/MS systems with Shimadzu HPLCs, and we are implementing a variety of other technologies and processes to increase data quality and speed and keep in accordance with industry standards.

BA LAB TEAM

The BA Lab has assembled an excellent team of scientists with 60+ combined years of pharmaceutical industry experience in bioanalytical, ADME, PK, In-Life, and pharmacology related fields.

Recently, Andrew Vick, PhD was appointed Executive Vice President and Senior Director of PDM. His contributions will further enhance our efforts in pharmacokinetics and drug metabolism for early stage drug development, where the identification of robust drug candidates is a critical first step in the development process. He brings extensive experience working within the biotechnology and pharmaceutical industry in the fields of toxicology, pharmacology, and drug disposition (PK/PD).

Seventh Wave President and Founder, Dr. John Sagartz (2nd from left), accepts the 2009 Outstanding Entrepreneur Award

Andrew Vick, PhDExecutive Vice President,Senior Director of PDM

SPEED, QUALITY, and RESPONSE TO CHALLENGES

The turnaround time for most single compound plasma PK studies is within 3 days with excellent data quality. (Statistics are based on studies completed between September 2008 and June 2009.)

Our Bioanalytical team has the problem-solving ability to respond to pharmaceutical and biotechnology clients' toughest drug discovery challenges.

XIC of +MRM (4 pairs): 271.2/155.2 amu from Sample 12 (Std. 0.5uM) of mdz_kcz_ r.wiff (Turbo Spray) Max. 4.3e4 cps.

0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2Time, min

0.0

5.0e5

7.4e5

In

te

ns

it

y,

c

ps

XIC of +MRM (4 pairs): 271.2/155.2 amu from Sample 12 (Std. 0.5uM) of mdz_kcz_ r.wiff (Turbo Spray) Max. 4.3e4 cps.

0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2Time, min

0.0

4.3e4

In

te

ns

it

y,

c

ps

1.91

XIC of +MRM (4 pairs): 325.8/291.3 amu from Sample 12 (Std. 0.5uM) of mdz_kcz_ r.wiff (Turbo Spray) Max. 7.4e5 cps.

0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2Time, min

0.0

5.0e5

7.4e5

In

te

ns

it

y,

c

ps

1.68

XIC of +MRM (4 pairs): 342.1/203.0 amu from Sample 12 (Std. 0.5uM) of mdz_kcz_ r.wiff (Turbo Spray) Max. 1.7e5 cps.

0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2Time, min

0.0

1.0e5

1.7e5

In

te

ns

it

y,

c

ps

1.66

XIC of +MRM (4 pairs): 531.9/490.3 amu from Sample 12 (Std. 0.5uM) of mdz_kcz_ r.wiff (Turbo Spray) Max. 1.3e4 cps.

0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2Time, min

0.00

1.00e4

In

te

ns

it

y,

c

ps

1.85

Column: Prevail C18_3u, 2.1x20, Lot No. 3506, PN: 43827 Filter: Javelin Column FilterMP-A: 0.1% FA in WaterMP-B: 0.1% FA in AcetonitrileWash-1: 1% FA in Acetonitrile: Water (15:85)Wash-2: AcetonitrileFlow Rate: 0.45mL/minTemperature: 35-40°CLoop size: 5uL

Representative Bioanalytical Method Representative In Vitro Data for CYP3A4 Inhibition Assay

Compound Name

Parent Ion

(amu)

Daughter Ion

(amu)Dwell Time

(ms) DP CE CXP

Tolbutamide 271.2 155.2 80 51 25 14

Midazolam 325.8 291.3 80 111 39 8

1-OH Midazolam 342.1 203 80 76 37 18

Ketoconazole 531.9 490.3 80 85 45 10

Time (min) % B

0.30 5

2.50 95

2.80 95

2.85 5

3.00 5

3.30 stop

Bioanalytical Lab of Seventh Wave

Percent Finished & Cumulative Percent Finished v.s. Study Turnaround Time (Days)

0

5

10

15

20

25

30

35

40

45

1 2 3 4 5 moreTime (Days)

% F

inis

he

d

0

20

40

60

80

100

120

Cu

mu

lati

ve

% F

inis

he

d

Midazolam Metabolic Stability in Human Recombinant CYP3A4 w/o Ketoconazole

0

0.5

1

0 5 10 15 20 30

Incubation Time (Minutes) (n=3)

Mid

azo

lam

Re

ma

inin

g (

µM

)

- NADPH + NADPH

Midazolam - Human CYP3A4Intrinsic Clearance w/, w/o Ketoconazole

y = -0.136x + 4.8092R2 = 0.9801

-2.2

-1.2

-0.2

0.8

1.8

2.8

3.8

4.8

5.8

6.8

0 5 10 15 20 25 30

Incubation Time (min)

Ln

(%

Rem

ain

ing

)

NADPH, w/o Ketoconazole NADPH, w/ ketoconazole

Midazolam - Human CYP3A4Intrinsic Clearance w/, w/o Ketoconazole

y = 122.64e -0.136x

R2 = 0.9801

0

20

40

60

80

100

120

0 5 10 15 20 25 30

Incubation Time (min)

% R

em

ain

ing

NADPH, w/o ketoconazole NADPH, w/ ketoconzole

Midazolam Metabolic Stability in Human Recombinant CYP3A4 w/ ketoconazole, w/ and w/o NADPH

0

0.5

1

0 5 10 15 20 30

Incubation Time (Minutes) (n=3)

Mid

azo

lam

Rem

ain

ing

(µ

M)

- NADPH, w/ ketoconazole + NADPH, w/ ketoconazole