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 [email protected]@musc.edu
Joachim Uys, Ph.D. PK project [email protected]