Transcript Drug Metabolism & Pharmacokinetics Core Facility

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
NOV-002, a myeloproliferative
agent
COOH
O
NH
NH
H2N
COONa
.
O
S
S
[Pt(NH3)2Cl2]
O
H2N
NH
COONa
NH
COOH
O
Phase II: Neo-adjuvant breast cancer trial
at MUSC
Pharmacokinetic Analysis
1 0 .0 0
ug/mL GSSG
A.
1 .0 0
Observed
Predict ed
0 .1 0
0 .0 1
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
0 .3 5
0 .4 0
0 .4 5
0 .5 0
Tim e
1 0 0 .0
ug/mL GSH
B.
1 0 .0
Observed
Predict ed
1 .0
0 .1
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
Ti m e
0 .3 0
0 .3 5
0 .4 0
0 .4 5
0 .5 0
Estimated pharmacokinetic parameters for NOV-002 (GSSG) and its
primary metabolite (GSH)
PK Parameter
GSH
GSSG
Co (ug/mL)
4.10
0.65
18.30
2.16
Cmax (ug/mL)
Tmax (hr)
0.16
Volume of distribution, V/F (L/kg)
0.15
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
Mortality
Saline
NOV-002
Cisplatin
Cisplatin + NOV-002
0/6*
0/6
1/8§
0/9
Elevated
Creatinine
0/6
0/6
6/7§
3/9
Abnormal
Renal Histology
0/6
0/6
7/7§
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 Toxicity
White Blood Cells

HM2 Hematology System (Abaxis)
Normal 6 x 109
Female +/+
x 109 cells/mL
3.00
5.00
2.80
4.00
2.60
3.00
2.00
2.40
1.00
2.20
0.00
2.00
1
Female
Male
Cisplatin +
NOV-002 -
Series1
Series3
1
+
+
Measure GSH/GSSG
Redox Potential
0
Eh (mV)
-180
-190
-200
-210
-220
50
100
150
200
250
Redox analysis:
-SH + TGTM
-S- TGTM
+
NOV-002
-S-SG
2D Graph 2
Y Data
% Free Sulfhydryls
110
100
90
*
80
*
*
70
*
60
50
0
5
10
30
X Data
Time vs % of Free SH
60
240
1440
Identification of Plasma Markers
following NOV-002 treatment
NOV-002 (25 mg/kg, iv)
Kda 0 ¼ ½ 1 4 24 h
150
100
A
B
75
50
C
37
D
WB: PSSG
WB: actin
Cost Recovery System:
•
•
•
•
•
•
•
•
Assay Development
PK Modeling
Plasma / Serum Assay
Organ toxicity analysis
Hematology (CBC)
GSH/GSSG levels
Protein Thiol Analysis
Breeze HPLC
– User Operated Training
– User Operation fee
– Over-night / weekend fee
$1,500 + Column
$50/ hr
$50 / sample
$35 / sample
$20 / sample
$25 / sample
$35 / sample
$ 50 / hour
$ 25 / hour
$18 / hour
Drug Metabolism and
Pharmacokinetics Core
Drug Discovery Building 420
843-876-2391
Danyelle Townsend, Ph.D., Director
Drug Discovery Building 420
843-876-2391
[email protected]
Yefim Manevich, Ph.D.,
[email protected]
Joachim Uys, Ph.D.
PK project manager
[email protected]