Open Access Central Laboratory’s: Application in Preclinical and Clinical Research Stages An Overview Rasmy E. Talaat, PhD Wyeth/Pfizer Fellow-Retired rasmy.talaat@gmail.com 1 ALEX: 29 November 2017
Drug MetabolismMission Statement The Mission of the Drug Metabolism Department is to conduct early DMPK studies to guide the selection, optimization and advancement of Discovery phase candidates, and to provide detailed metabolic disposition characterization of Development compounds in support of clinical development and regulatory submissions - Both Small Molecules and Proteins from all therapeutic areas ALEX: 29 November 2017 2
Drug Metabolism Strategic Advantages Organizational Structure Why it works: – Leadership. – Preclinical regulatory considerations to early support clinical development strategies – Efficient processes and workflow permits timely dissemination of data – Maximizes efficiencies through harmonization, standardization and automation of routine methodological processes – Innovative approaches using state-of-the-art technology to characterize potential development candidates Consequence: – Improved productivity, elimination of redundant studies to produce better quality compounds advancing into Development via consensus on decisions to advance or terminate a NCI. – Proven productivity track record of supporting 8+ INDs and 2-3 NDA per year ALEX: 29 November 2017 3
LEADERSHIP • Experience: UW, ABC Labs, Covance, Parke Davis, Wyeth/Pfizer • Open and Free Training across departments as well as Clients • UMBC Undergraduate and Graduate students Hands on Training • Training 200+ users • Customized levels of training • Independent operating research with Free guidance • Free consultation to external MS users • Intra-, Inter-, and External Collaboration between team researchers
Drug Metabolism - Strategic Advantages Staff Resources and Technology Platforms • Centralized, highly integrated, full-service Drug Metabolism laboratories • Highly skilled, flexible, diverse and multi-disciplined workforce provides a complement of expertise that can be used to address DMPK/Safety/Toxicology in Discovery and Development – 39% of Staff have Ph.Ds, with expertise in a range of disciplines, including Drug Metabolism , PK , Mass Spectrometry, Toxicology, Molecular Biology and Analytical Chemistr y • Laboratories equipped with sophisticated, state-of-the-art instruments capable of supporting both Discovery and Development phase studies from all therapeutic areas and platforms ALEX: 29 November 2017 5
Mass Spectrometry Principle of operation A Mass spectrometer is a “ Molecule Smasher ” Measures molecular and atomic masses of whole molecules, molecular fragments by generation and detection of the corresponding gas phase of ionized molecules in vacuum , separated according to their mass-to-charge ratio (m/z) with the aim of one or more of the following: • Molecular weight determination • Structural characterization • Gas phase reactivity study • Qualitative and Quantitative analysis of components in a mixture. 6 ALEX: 29 November 2017
Drug Metabolism Capital Equipment Mass Spectrometry, Automation, NMR • Mass Spectrometers - Total of 31 in Drug Metabolism Department (35 users-40% PhD) – Quantitative Analyses : Seventeen Triple Quadrupole MS – Metabolite Identification : Fourteen High Resolution Mass Spectrometers e.g. LTQ Orbitrap, Q-TOF-2, …… . • LC-MS/MS-NMR 600 MHz Bruker NMR Magnet (1 PhD and 1 MSc) • IT (MSc) • Automation (3 MSc) – Eight Liquid Handlers e.g. Tomtec Quadra-4, Tecan Freedom, Packard Multiprobe II, …… . ALEX: 29 November 2017 7
Major Functions of Mass Spectrometry Discovery/Development Metabolism • In vitro Metabolite Profiling: • Provide metabolite profiles for drug candidates generated from in vitro systems (liver microsomes + Cytosole, Hepatocytes) to assist BETTER drug candidate design. • In vivo Metabolite Profiling of PK/TK Plasma : Associate in vitro metabolism with the in vivo • Examine the correlation of metabolite with adverse effects. • Reactive Metabolite Identification: • Assess potential bio-liabilities by detection of reactive metabolite s using trapping techniques (e.g. GSH, KCN, Dipeptide) along with the recommendations to minimize/avoid the formation of such reactive metabolite in drug design . 8 ALEX: 29 November 2017
DPK 2008 Productivity • Small Molecules – Total Studies 1969 – Neurosciences 583 – CVMD 470 – Inflammation 353 – Oncology 419 – WH&B 144 • Proteins – Total Studies 50 ALEX: 29 November 2017 9
Pre-clinical PK Productivity # of Studies 2250 1969 Total Studies 2000 1787 1665 Contracted Studies 1750 1500 1284 1250 966 1000 741 750 488 500 170 122 98 250 77 59 47 0 0 2002 2003 2004 2005 2006 2007 2008 # of FTEs has been reasonably constant over the years ~23 ALEX: 29 November 2017 10
APPLICATIONS OF THE MASS SPECTROMETRY/AUTOMATION GROUP IN ADME-SAFETY of DRUG DISCOVERY-DEVELOPMENT STAGES DISCOVERY METABOLISM DISCOVERY PHARMACOKINETICS • CYP inhibition reversible IC 50 cocktail assay • Protein precipitation • CYP inhibition mechanism-based screen (conventional and filter plate) • Liquid-liquid extraction • CYP inhibition kinetic studies: K i or K I , (conventional and solid- k inact supported) On-line LC/MS automated systems • Reaction phenotype (cold studies) HIGH THROUGHPUT INCUBATION • Solid phase extraction • K m substrate depletion or K m , V max AND EXTRACTION TOOLS o (microplate based and on-line) determinations Variable channel liquid handlers • Method Development & • Intrinsic clearance and metabolite ID 96-channel pipettors Optimization Robotic arms and rails • Trapping reactive intermediate metabolites • Biomarker assays PROTEIN BINDING DEVELOPMENT METABOLISM • Equilibrium dialysis (hot and cold studies) • Reaction phenotype (hot studies) • Ultracentrifugation (hot and cold studies) • Plasma and whole blood stability • Ultrafiltration (hot and cold studies) • 384 well UGT assay Plasma, Microsomes and Brain Tissue for better • Clinical BA Studies First in Human dose projections and improved PK- PD in CNS
TYPICAL FACT SHEET L. Leung May 13, 2009 PHARMACOLOGY PHYSICOCHEMICAL PHARMACOKINETICS In vitro IC 50 PROPERTIES Balb/C Mouse Enzymatic: 0.9 nM m.w. 437 logP: 3.13 iv 2 mpk WBC Stat PO 4 : 59 nM CL = 12 mL/min/kg Permeability (x 10-6 cm/sec) Whole blood: 580 nM Vss = 2.1 L/kg; T 1/2 = 2.3 h Caco2: A-B 14.9 (DSM) ER ~2 P-gp inhibition IC 50 : 10 – 100 M In vivo po 10 mpk (MC/TW) Mouse IL-2/IFN g : ED 50 : 1 mg/kg AUC last = 5200 h*ng/mL Solubility C max = 750 ng/mL; F = 36% DTH mode ED 50 : 10 mg/kg QD <LOQ (pH 7.4), 0.002 mg/mL (pH 1), SD Rats 0.019 mg/mL in MC/TW Mouse CIA model: MED 3 mg/kg BID iv 2 mpk 0.32 mg/mL in 10% TPGS 3 mg/kg single dose Cmax: 217 ng/mL CL = 16 mL/min/kg BCS: 2 AUC last =2089 ng.hr/mL Vss = 1.8 L/kg; T 1/2 = 1.4 h Protein Binding Extrapolated AUC at MED = 4178 ng.hr/mL po 10 mpk (10% TPGS) >97% human; 92% rat and AUC last = 1056 h*ng/mL SAFETY 91% mice (PCOP) Moderate cytotoxicity (IC 50 = 6 M) C max = 240 ng/mL; F = 9.4% Unbound 3.8% human, 4.1% DBA Ascending po dose (10% TPGS) mouse, 6.3% rat, 4.6% dog and 3.9% No major findings in 7-day EDS AUC last (h*ng/mL) = 9276 (100 mg/kg), monkey (DSM) assessemnt in CD-1 mice at 10 and 14231 (300 mg/kg), 24517 (1000 Blood/Plasma: 1 (r), 0.7 (h) 100 mg/kg/day METABOLISM mg/kg) hERG (IC50) Stability (DSM) Dogs Weak activity Hepatocytes CL int (µL/min/10^6 cells): iv 2 mpk (27% inhibition at 10 M) CD-1 Mouse(LM)/Rat/ dog/ monkey/ CL =25 mL/min/kg Ames human Vss = 1.2 L/kg; T 1/2 = 0.8 h Negative 8/4.0/4.6 / 63 / 9.4 po 10 mpk (10% TPGS) PHARMACOKINETICS CYP450 Inhibition AUC last = 2426 h*ng/mL F = 35% Monkeys DSM IC 50 (µM) C max = 1556 ng/mL iv 2 mpk Ascending po dose (10% TPGS) 1A2: NI 2A6: NI 2C9: 4 2C19: 24 CL =25 mL/min/kg AUC last (h*ng/mL) = 1213 (30 mg/kg), 2D6: 47 3A (MDZ): NI (TST): 56 Vss = 0.95 L/kg; T 1/2 = 0.6 h 6754 (100 mg/kg), 8988 (300 mg/kg) Not a MBI po 10 mpk (10% TPGS) DDI: possible for CYP2C9 AUC last = 35 h*ng/mL F = 0.4% Induction: Reporter assay at 2 uM: 2.4 C max = 6 ng/mL 12 fold (CYP3A4), 3 fold (CYP1A2) ALEX: 29 November 2017
The Bioanalytical Center • To establish an International Bioanalytical Center . • To provide superior quality analytical data with unsurpassed turn-around time to support Preclinical and Clinical Research and to enhance Life Sciences Research . • Create highly versatile scientific community • Support and Advance e.g. Medical and Clinical studies, Academic research, Analytical research, and • Drug Discovery and Development • Economically highly rewarding • Generate Steady Income to self support 13 ALEX: 29 November 2017
Recommend
More recommend
