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 Phase II: Neo-adjuvant breast cancer trial at MUSC

Pharmacokinetic Analysis

B. A. ug/mL GSSG ug/mL GSH

Estimated pharmacokinetic parameters for NOV-002 (GSSG) and its primary metabolite (GSH) PK Parameter GSH GSSG C o (ug/mL) C max (ug/mL) T max (hr) Volume of distribution, V/F (L/kg) K01 (absorption rate constant), hr K10 (elimination rate constant), hr AUC 0-30min (ug.h/mL) Clearance, CL/F (L/hr/kg)

Evaluation of Organ Toxicity

Table 2. Mortality and Nephrotoxicity of Cisplatin in Mice Elevated Abnormal Mortality Creatinine Renal Histology Saline0/6*0/60/6 NOV-0020/60/60/6 Cisplatin1/8§ 6/7§7/7§ Cisplatin + NOV-0020/93/93/9 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) Female Male x 10 9 cells/mL Normal 6 x 10 9 Female +/+ Cisplatin + + NOV

Measure GSH/GSSG Redox Potential E h (mV)

Redox analysis: -S- TG TM -SH NOV TG TM + -S-SG % Free Sulfhydryls * * * *

A B C D Kda NOV-002 (25 mg/kg, iv) 0 ¼ ½ 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 Danyelle Townsend, Ph.D., DirectorYefim Manevich, Ph.D., Drug Discovery Building Joachim Uys, Ph.D. PK project manager