N
Drug Metabolite Identification Services for FDA
IND/NDA Safety Assessments
extcea
Nextcea performs drug metabolism assays to identify and quantitate drug metabolites in
animals and humans in compliance with FDA guidance for safety testing of drug metabolites
Metabolite Identification Services for Safety Assessment
Biotransformation occurs via Phase I (aliphatic and aromatic oxidation, N-oxidations, dealkylations, reductions...)
and Phase II (glucuronidation, glutathione, sulfation conjugations...) metabolism. The FDA encourages “the
identification of differences in drug metabolism between animals used in nonclinical safety assessments and humans
as early as possible during the drug development process1.” Animals used in drug safety assessment should have
systemic exposure to drug metabolites at least comparable to the major drug metabolites observed in humans.
In Vitro Assays
Phenacetin metabolic pathway
O
O
H3 C
HN
O
Nextcea offers a full range of in vitro services for drug metabolite
identification and across-species profiling including incubations
with hepatocytes, liver microsomes, S9 fractions, and recombinant
human enzymes such as CYP450 isozymes (1A2, 3A4, 2D6,
2C8, 2C9, 2C19...), flavin monoxygenase isozymes, and
UDP glucuronosyl transferases isozymes (UGT1A1, 1A10, 1A3,
1A4...).
O
HO
H3 C
S
HN
O
O
Chemical Formula: C 8H9NO5S
Exact Mass: 231 .02 01
Phenacetin
Chemical Formula: C 18H22N 2O4
Exact Mass: 33 0.1 580
M4 (MO, D, MK, H)
Sulfation
Dealkylation
O
H3 C
HN
OH
Acetaminophen
Chemical Formula: C 8H9NO2
Exact Mass: 15 1.0 633
M1 (MO,D, MK, H)
Glucuronidation
O
O
H
O
O
H3C
C
O
OH
S
HN
H
HO
OH
HO
H
Chemical Formula: C 14H17NO8
Exact Mass: 3 27.0954
M2 (MO, D, MK, H)
In Vivo Assays
OH
HN
O
H3C
HN
Gluthathione
Conjugation
O
O
NH2
HN
OH
O
O
Chemical Formu la: C 18H 24N4O 8S
Ex act M ass: 456.131 5
M3 (MO, D, MK, H)
Nextcea also performs in vivo metabolite identification services
across any animal species the client prefers. Animal and human
biological in vivo samples can be provided by the client for
metabolite identification. Alternatively, test compounds can be
submitted directly to Nextcea and the animal dosing and sample
collection will be coordinated at a partner animal facility.
Metabolite Synthesis
Nextcea synthesizes potential metabolites for structure confirmation and quantitation. The FDA recommended the
synthesis and dosing of suspected toxic metabolite(s) to animals if they are found exclusively or in disproportionately
higher levels in humans1.
Study Design & Across Species Metabolite
Analysis
Nextcea provides expertise in in vivo drug metabolism
study design to meet the clients’ drug development needs.
Data is presented in a detailed report including drug
metabolic pathways with across species comparison.
Nextcea can conduct studies to confirm the structures
of metabolites and develop assays to quantitate
metabolites and determine exposure in humans and
animals. Additionally, drug metabolism study results
can be correlated with toxicological outcomes if
desired by the client.
About Nextcea, Inc.
Admetry
File
Pharmacokinetics Drug Metabolism User Database Help
Metabolite ID
Exact Mass:
511.215I
5I onization Method: (M+H)+
Drug Metabolism
Calculated Metabolite Results
Phase I Metabolites
One
Reaction
Hydrogenation
Hydrogenation, Oxidation, Epoxidation (Aromatic &
Aliphatic )
S-Oxidation
P-Oxidation (S replacement)
Hydrolysis (Ester, O=C-O-CH3 -> OH )
Hydrolysis (Ester, O=C-O-CH2-CH3 -> O=C-OH )
Hydrolysis (Amide, O=C-NH-CH3 -> OH )
Hydrolysis (Amide, O=C-NH-CH2-CH3 -> NH2 )
Two
Reaction s
Two
Combined
Reaction s
498.207 1
484.191 4
514.20 2
528.217 7
544.212 6
Three
Combined
Reactions
Hydroxylation, Oxidation, Epoxidation (Aromatic & Aliphatic )
Mass: 15.9949
Hydroxylation of Aliphatic Carbon:
O
O
O
H3CS
NH
O
HOCH 2
S
HN
O
Dehydrogenation
NH
HN
O
Tolbutamide
Dehydrogenation (alcohol oxidation to aldehyde, double
bond formation)
O
O
CH 2CH 3
HN
CH 2CH 3
HN
CH 2CH 2CH 2CH 2CH 3
Dealkylation (N, O, S de-methylation)
CH 2CH 2CH 2CHCH3
OH
O
N
H
O
O
N
H
O
Nextcea employs proprietary bioinformatic
software (Admetry®) to identify metabolites across
different species.
Nextcea, Inc. is a drug development service company dedicated to
optimizing efficacy and minimizing toxicity in all phases of drug
development.
Nextcea integrates cross-species ADME and
biomarker studies with traditional PK/PD and TK/TD. In-house
platforms include mass spectrometry LC-MS and LC-MS/MS
(API-6500s and TripleTOF 6600), and proprietary pharmacokinetic
and drug metabolism bioinformatics (Admetry®).
500 W. Cummings Park, #4550
Woburn, MA 01801
781-457-4010
[email protected]
Hirohide Mimura (三 村 博 英):
[email protected]
1 Guidance for Industry: Safety Testing of Drug Metabolites. Pharmacokinetic Subcommittee at the Food and Drug Administration.