GVK’s In-vitro ADME services offer a portfolio of assays for investigating: metabolism, distribution and toxicity, permeability, solubility & physicochemical properties, for more info please visit http://www.gvkbio.com/adme.html

1. CMYK
ADME SERVICES
Our In-vitro ADME screening service offers a portfolio of Solubility
assays for investigating: metabolism, distribution and toxicity.
permeability, solubility & physicochemical properties, Solubility is one of the most important physicochemical
GVK BIO delivers consistent, accurate compound data properties. We determine equilibrium solubility by dried DMSO
with cost-efficiency. method. We can also determine solubility in aqueous buffer (pH
1-9), organic solvents (DMSO, Ethanol, etc), formulations and
In-vitro ADME Capabilities excipients by pION/Multiscreen/HPLC/UV/MS/MSMS methods.
Physicochemical studies
• Log D/Log P
Protein Binding
• Solubility(Kinetic/Equilibrium)
In-vitro binding studies with plasma have proven to be a valuable
• Chemical stability tool for predicting In-vivo protein binding. We determine the
protein binding by both ultra filtration and rapid equilibrium
• Biological matrix stability (serum/ plasma/
dialysis. Using RED device we determine protein binding in
microsomes/ blood /hepatocytes/tissue homogenates)
microsomes. Plasma and tissue homogenate across various
Absorption/Distribution Assays species (Rat/mice/human &dog).
• Caco2 and PAMPA permeability assay
• Pgp substrate / inhibitor assay
Caco2 Permeability Assay
• Protein binding
Caco2 cells are the most frequently
• Blood/Plasma Partitioning Ratio used In-vitro models to assess
intestinal permeability. Permeability
Metabolism/Excretion across Caco2 cell monolayer is used
• Half life/clearance determination using to predict human permeability of drug
microsomes/Hepatocytes /S9 fractions/Cyp across species candidates, to perform in-depth
(Human/Rat/Mouse/Dog/ monkey) mechanistic and absorption studies,
to study the effects of transporters on
• CYP Inhibition (CYP1A2, CYP2C9, CYP2C19, permeability. We can determine
CaCo-2 Cells
CYP2D6, CYP3A4) apparent permeability (Papp) / efflux Semi-permeable
• Pathway determination (Phase I and Phase II) ratio / Unidirectional / Bidirectional membrane
by LCMS.
• Metabolite identification using Microsomes / Hepatocytes
and
• Characterization of potential metabolites using microsomes
and Hepatocytes across species (Human/Rat/Mouse/Dog/ PAMPA
monkey) (Parallel artificial membrane permeability assay)
Log D/Log P The Parallel Artificial Membrane Permeability Assay (PAMPA)
assay is used as an in-vitro model of passive, transcellular
Log D is determined by the shake-flask method, by dissolving
some of the solute in a volume of octanol and water/buffer and permeability. As well as for the prediction of oral absorption and
measure the concentration of the solute in each solvent. Log D is brain penetration. Effective permeability (log Pe) may be
determined by HPLC-UV with confirmation by mass. measured by pION/Multiscreen/LCMS.
CMYK

2. CMYK
ADME SERVICES
Microsomal Microsomal Stability
CYP2D6 inhibition by Furafyline CYP2D6 inhibition by Qunidine
Stability A
B
C
1 2 3 4 5 6 7 8 9 10 1112
D
96 - Well Plate
Metabolic stability plays E
F
G
H
110 EC50 3.092e-006 80 EC50 1.015e-008
an important role in the 90 R2 0.9968 70 R2 0.9900
Chemicals
success of drug 70 60
candidates. First pass 50
% Inhibition
% Inhibition
50
40
metabolism is one of Add Hepatocytes 30
30
the major causes of or Microsomes 10
20
0
poor oral bioavailability 10
-10
and short half life and -8 -7 -6 -5 -4
0
-10 -9 -8 -7 -6 -5
the study influences
Incubate
both oral bioavailability Log drug Con [M] Log drug concentration [M]
and half life. The half life
/ clearance / % Extract
metabolized can be CYP2D6 inhibition by Sulphaphenazole CYP3A4 - Ketoconazole
determined in
microsomes / LCMS Analysis
S9 fractions / 35 EC50 6.372e-007
110
EC50 1.884e-007
100
hepatocytes. 30
2
R 0.9431 90
R2 0.9979
80
25
% Inhibition 70
20 60
Hepatocyte metabolic stability
% Inhibition
50
15 40
10 30
Per orally administered drug may undergo first-pass metabolism 5
20
10
which influences the pharmacokinetic properties such as the 0 0
-9 -8 -7 -6 -5 -4 -10
clearance, the half-life or the bioavailability. Cryopreserved -10 -9 -8 -7 -6
hepatocytes are used to calculate half life/clearance/% parent Log drug Concentrations [M] Log drug concentration [M]
compound remaining.
Human hepatocytes metabolic stability Metabolite identification using
Light Sight Software
% Parent compound remaining
Metabolic stability and the identification of formed metabolites has
100
become an important tool. In-vitro metabolite identification &
90
80 characterization of the test compounds is determined using Q
70 trap with light sight software.
60
50
40
30 Bioanalysis
20
10
LC-MS/MS
0 ?
3200 QTRAP
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API3200
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mil
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min
no
7-E
no
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Ate
API4000
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Pro
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HPLC
Compound details ? RRLC with PDA
Agilent 1200
?
Shimadzu Prominence with UV
? RRLC with HTS PAL
Agilent 1200
CYP Inhibition Others
CYP inhibition occurs either as reversible inhibition, quasi- ? (Molecular Devices)
Spectramax
irreversible inhibition or irreversible inhibition. CYP inhibition is a
fluorescent based/LCMS assay. Specific isoforms of CYP ?
BMG Polarstar
(CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) are ? Handler (Tomtec)
Qudra4 Liquid
used to determine the inhibition (IC50).
GVK Biosciences Private Limited
28A, IDA, Nacharam, Hyderabad 500 076, India. T 91 40 66281823
F 91 40 6628 1505 E-mail: bdbio@gvkbio.com Website: www.gvkbio.com
CMYK