In Vitro ADME

Whether you are in the early stages of discovery or preparing to submit an NDA, our comprehensive platform of in vitro services provides you with the quality data you need to predict pharmacokinetics (PK) and pharmacodynamics (PD) in vivo. Offering a comprehensive list of assays, from early discovery to definite regulatory designs, supported by our bioanalytical services unit for method development and validation, our in vitro ADME services are available for both small and large molecule drugs, and can be tailored to fit the specific requirements of your program.

Specific Capabilities: in vitro ADME

Physical Chemistry

  • Pharmaceutics: Solubility, pKa, logD, logP

Absorption

  • Permeability: Caco-2, MDCK, PAMPA
  • Efflux transporters BCRP, P-gp, BSEP, MRP2, MRP3, MRP4 as membrane vesicles
  • Transfected and double transfected MDCK II cells (BCRP, P-gp, BCRP±OATP2B1)

Distribution

  • Red blood cell partitioning
  • Protein binding: Dialysis, ultracentrifugation, etc.
  • Tissue binding

Metabolism

  • Matrix stability: Tissue, plasma, SGF/SIF, buffer
  • Metabolic stability: Microsomes, S9, hepatocytes, CYP phenotyping
  • Metabolite ID: Structure determination and quantitation

Drug-drug interaction

  • CYP Inhibition/Induction: IC50, reversible inhibition, TDI, KI/Kinact, activity and/or mRNA
  • Transporters OAT1, OAT3, OATP1B1, OATP1B3, OCT1, OCT2, PEPT1, 2, MATE1, 2K, NTCP, ASBT as cell-based assays

Assay Automation

  • Four liquid handling systems (Tecan, Biomek) for ADME assay automation (latest addition Tecan fluent 1080)
  • Rigorous validation and trending of each automated assay.
  • Currently automated assays include:
  • Permeability assays
  • Metabolic stability
  • Transporter assays
  • Cyp Inhibition
  • Dilutions for bioanalytical method development

Learn about our 10mg DMPK Discovery Package

Solution I: We know the target (assume the brain is the target)

Assay

Study Design

Compound Distribution(mg)

In vitro

ADME

Phase Ⅰ

MMS

Liver microsome stability (CD-1 mouse and Human)

2.0

PPB

Plasma protein binding - HTD (CD-1 mouse and Human)

BHB

Brain homogenate binding (CD-1 mouse)

MDR1

Bidirectional permeability in MDR1-MDCK II cells

DDIM

CYP inhibition for 5-in-1 cocktail substrates

KS

Aqueous solubility (Kinetic)

Log D

LogD measurement at pH 7.4

Phase Ⅱ (Option)

HMS

Stability in hepatocytes (CD-1 mouse and Human)

Met ID

In vitro Met ID in liver microsomes (SD rat, Human, Beagle dog,

Cyno monkey and CD-1 mouse)

In vitro Met ID in hepatocytes (SD rat, Human, Beagle dog, Cyno

monkey and CD-1 mouse)

PLS

Stability in plasma (CD-1 mouse and Human)

Caco-2

Bidirectional permeability in Caco-2 Cells

Efflux potential evaluation in Caco-2 Cells

TDI

CYP time dependent inhibition (IC50 shift, 5 CYPs, 6 substrates)

pKa

pKa measurement (spectrometric method)

Mouse PK Study

· 9 male mice (CD-1/C57BL/6), n=3/route

· Group 1& Group 2: Single compound administered via 2 routes (IV+PO), 7-8 blood sampling time points up to 24 hours post dosing per animal

· Group 3: Brain perfusion, terminal sacrifice, collect brain and CSF

· Method development and 48 plasma samples +3 brain samples

+3 CSF samples bioanalysis with LC-MS/MS

· PK data submission in excel report

2.0

Mouse PD Study

· Choose appropriate animal model and the dosage of PD study depends on PK parameters

Group 1: 6 males PO (normal)

Group 2: 6 males PO (vehicle)

Group 3: 6 males PO (positive control)

Group 4: 6 males PO, Plasma, CSF and brain (low) Group 5: 6 males PO, Plasma, CSF and brain (middle) Group 6: 6 males PO, Plasma, CSF and brain (high)

· PK data submission in excel report

6.0

 

Solution II: We don’t know the target

Assay

Study Design

Compound Distribution(mg)

In vitro

ADME

Phase Ⅰ

MMS

Liver microsome stability (CD-1 mouse, SD rat and Human)

2.0

PPB

Plasma protein binding - HTD (CD-1 mouse, SD rat and Human)

MDR1

Bidirectional permeability in MDR1-MDCK II cells

DDIM

CYP inhibition for 5-in-1 cocktail substrates

KS

Aqueous solubility (Kinetic)

Log D

LogD measurement at pH 7.4

Phase Ⅱ (Option)

HMS

Stability in hepatocytes (CD-1 mouse, SD rat and Human)

Met ID

In vitro Met ID in liver microsomes (SD rat, Human, Beagle dog,

Cyno monkey and CD-1 mouse)

In vitro Met ID in hepatocytes (SD rat, Human, Beagle dog, Cyno

monkey and CD-1 mouse)

PLS

Stability in plasma (CD-1 mouse, SD rat and Human)

Caco-2

Bidirectional permeability in Caco-2 Cells

Efflux potential evaluation in Caco-2 Cells

TDI

CYP time dependent inhibition (IC50 shift, 5 CYPs, 6 substrates)

pKa

pKa measurement (spectrometric method)

Rodent PK

Mouse PK study

· 6 male mice (CD-1/C57BL/6), n=3/route

· Group 1& Group 2: Single compound administered via 2 routes (IV+PO), 7-8 blood sampling time points up to 24 hours post dosing per animal

· Method development and 45 plasma samples bioanalysis with LC-MS/MS

· PK data submission in excel report

8.0

Rat PK study (Option)

· 6 male SD rats, n=3/route

· Group 1& Group 2: Single compound administered via 2 routes (IV+PO), 7-8 blood sampling time points up to 24 hours post dosing per animal

· Method development and 45 plasma samples bioanalysis with LC-MS/MS

· PK data submission in excel report

End-to-End ADME Solutions

When you engage the Laboratory Testing Division as your testing platform, you can be confident our team of experts will take your compound from start to finish, navigating all of the complexities in between. Our in vitro services span from discovery through definitive ADME, delivering the data-driven support you need to advance your development efforts and reach the ultimate goal of providing therapeutic value for patients.

 

Discovery ADME

IND-Enabling ADME (Early ADME)

NDA-Enabling ADME (Definitive ADME)

Physicochemical Characterization

X

X

X

Permeability & Solubility

X

X

X

Solution-Phase Stability

X

X

 

Metabolism and Clearance

X

X

X

CYP Inhibition

X

X

X

CYP Induction

 

X

X

Protein Binding (Plasma, Brain or Tissue Homogenate)

X

X

X

Blood-Plasma Partition

 

X

X

Transporter Drug-Drug Interactions (inhibition and substrate)

 

X

X

Metabolite Identification and structure elucidation

 

X

X

Enzyme Phenotyping (CYP, UGT, MAO, AO, SULT, etc)

 

X

X