Drug Metabolism and Pharmacokinetics Core Goal: To provide comprehensive pharmacology support for...

Drug Metabolism and Pharmacokinetics Core

Goal: To provide comprehensive pharmacology support for preclinical and clinical research

Services:

• Pharmacokinetic studies: Quantify drug levels in serum/plasma, tissue or cells

• Evaluation of Organ Toxicity: Blood chemistry analysis for liver, kidney and bone marrow toxicity

• Hematology: Measure complete blood counts• Measure GSH/GSSG• Redox analysis: Detection of protein thiol

status in cells, tissue, serum and plasma.• Education and training

H2N

COOH

O

NH

O

NH

S

S

O

NHNH

O

COOH

H2N

COONa

COONa

. [Pt(NH3)2Cl2]

NOV-002, a myeloproliferative agent

Phase II: Neo-adjuvant breast cancer trial at MUSC

Pharmacokinetic Analysis

B.

A.

0.1

1.0

10.0

100.0

0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50

Time

Observed

Predicted

0.01

0.10

1.00

10.00

0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50

Time

Observed

Predicted

ug/

mL

GS

SG

ug/

mL

GS

H

Estimated pharmacokinetic parameters for NOV-002 (GSSG) and its primary metabolite (GSH)

PK Parameter GSH GSSG

Co (ug/mL) 4.10 0.65

Cmax (ug/mL) 18.30 2.16

Tmax (hr) 0.16 0.15

Volume of distribution, V/F (L/kg) 42.61

K01 (absorption rate constant), hr-1 6.42

K10 (elimination rate constant), hr-1 5.81 6.40

AUC 0-30min (ug.h/mL) 6.37 1.18

Clearance, CL/F (L/hr/kg) 273.15

Evaluation of Organ Toxicity

Table 2. Mortality and Nephrotoxicity of Cisplatin in Mice

Elevated AbnormalMortality Creatinine Renal Histology

Saline 0/6* 0/6 0/6NOV-002 0/6 0/6 0/6Cisplatin 1/8§ 6/7§ 7/7§Cisplatin + NOV-002 0/9 3/9 3/9

•Data are the number of mice who died, had elevated plasma creatinine (>0.2 mg/dL) or abnormal renal histology over the number of mice treated.§ Statistically significant differences between treatment groups were detected with a Fisher Exact Test, P<0.01.

Saline NOV-002

Cisplatin Cisplatin + NOV-002

Hematology:

Cisplatin Induced Bone Marrow ToxicityWhite Blood Cells

HM2 Hematology System (Abaxis)

Female Male

x 1

09 c

ells

/mL

Normal 6 x 109 Female +/+

Cisplatin + + NOV-002 - +

Measure GSH/GSSGRedox Potential

Eh (

mV

)

Redox analysis:

-S- TGTM -SH

NOV-002

+ TGTM

+

-S-SG

% F

ree

Su

lfhyd

ryls

**

*

*

A B

C

D

Kda150100

75 50

37

NOV-002 (25 mg/kg, iv) 0 ¼ ½ 1 4 24 h

WB: PSSG

WB: actin

Identification of Plasma Markers following NOV-002 treatment

Cost Recovery System:

• Assay Development $1,500 + Column• PK Modeling $50/ hr• Plasma / Serum Assay $50 / sample• Organ toxicity analysis $35 / sample• Hematology (CBC) $20 / sample• GSH/GSSG levels $25 / sample• Protein Thiol Analysis $35 / sample• Breeze HPLC

– User Operated Training $ 50 / hour– User Operation fee $ 25 / hour– Over-night / weekend fee $18 / hour

Drug Metabolism and Pharmacokinetics Core

Drug Discovery Building 420843-876-2391

Danyelle Townsend, Ph.D., Director Yefim Manevich, Ph.D., Drug Discovery Building 420 manevich@musc.edu843-876-2391townsed@musc.edu

Joachim Uys, Ph.D. PK project manageruys@musc.edu