We have added to our extensive selection of metabolism and related →ADME/DMPK services new assays for conjugation metabolism analysis (also known as phase II metabolism). Conjugation metabolism, in addition to the redox metabolism (phase I metabolism, mainly by CYP P450 enzymes) forms the major biotransformation machinery of endogenous and xenobiotic (drug) compounds.

The importance of studying the conjugation metabolism is also added to the very recent (Feb-2012) FDA Draft Guidance → Guidance for Industry: Drug Interaction Studies — Study Design, Data Analysis, Implications for Dosing, and Labeling Recommendations, 2012.

Whilst the →phase I metabolism analytics (CYP identification, inhibition, induction, suppression) is currently routinely done in drug discovery programs, good scientific expertise in phase II conjugation metabolism is much less commonly available.

We are offering a spectrum of analytics and scientific expertise in phase II conjugation metabolism

We have started with the most common conjugation metabolism enzymes, the UGTs (Uridine 5′-diphospho-glucuronosyltransferases) with our recent staffings and the currently available analytical assays include:

1) UGT-Pilot – General assessment of glucuronidation for the drug compounds

This study is devised to detect if glucuronidation happens for the studied compounds as part of its biotransformation. It does not pinpoint which UGT enzyme is responsible for the glucuronidation. Should this study indicate involvement of UGTs in the drug metabolism, the recommended followup is to identify the metabolizing UGT(s) (see also the flowchart in the FDA new draft guidance).
The compound is incubated in test system, typically liver microsomes (different species available). Formation of glucoronides is detected by validated UPLC-MS/MS analytics.

2) UGT-Id – Pinpointing the specific UGT enzyme responsible for the glucuronidation of the compounds

This study is meant to identify the UGT enzyme(s) responsible for the glucuronidation and thus it is most often used as UGT-Pilot follow-up study. It is thus analogous to →our CYP identification assay (see this page for more). We currently have analytical assays for the 10 most common human UGT enzymes, including UGT1A1 (see also the FDA new draft guidance).
The compound is incubated with insect cell expressed human UGT enzymes (Supersomes). Formation of the gulucorinides is detected by UPLC-MS/MS analytics.

3) Inhib-UGT – Detection of the UGT inhibition by the studied compound(s)

This study is for detecting whether the compounds inhibit the activity of a specific UGT enzyme. This is analogous to →our CYP inhibition assay (see this page for more). We are detecting the potency to inhibit any of the 10 most common human UGTs.
The compound is incubated with a specific reporter substrate and insect cell expressed human UGT enzymes (Supersomes). The inhibition is detected by lower amounts of the glucuronidation on the reporter substrates, as detected by a specific UPL-MS/MS analytics.

It should be noted that a compound can inhibit the UGT enzyme activity, even though it would not be metabolized itself by the enzyme, and thus Inhib-UGT study is recommended to be doen irregardless of the results from the UGT-Pilot and UGT-Id studies.

Other phase II conjugation metabolism assays

These UGT specific assays will soon be followed with analytical assays for other classes of phase II metabolizing enzymes as well as various phase II enzyme induction assays.

Promotional offer for phase II conjugation metabolism studies

We offer a special 20% discount on phase II conjugation metabolism studies, which have been ordered during March-April, 2012. The promotional offer also includes 25% discount on CYP inhibition and CYP identification studies, when ordered together with phase II conjugation metabolism studies. This promotional offer will also include the other phase II conjugation metabolism assays that will be launched during this period.

Please contact us for more information.

Cytochrome P450 enzymes (CYPs) have a central role in drug metabolism. Drug candidates inducing CYP expression or enzyme activity can potentially increase rate of metabolism and under-exposure of co-administered drugs. Strong CYP induction can be also a sign of hepatotoxicity.

We are pleased to offer a novel solution to study CYP and other drug metabolism related genes activity – TRAC. TRAC is a novel high-throughput solution for multiplexed expression analysis. We offer TRAC in collaboration with →Plexpress, the developers of the patented TRAC method and located next door to SBW in Viikki, Helsinki. Our combined service allows more efficient screening of gene expression induction of CYPs and other ADME related transcripts in human primary hepatocytes or HepaRG cells by the drug compounds. The gene expression analysis can also be combined with LC/MS-MS determination of →CYP metabolites as well as →CYP identification and CYP inhibition.

Compared to other methods, TRAC offers
• Significant cost savings in gene expression screening compared to qPCR
• All relevant CYP genes readily available as a multiplexed assay
• Multiplexing capacity of up to 30 genes/sample
• Reliable detection: excellent repeatability with CVs <10%, good correlation with qPCR
• Reliable normalization: housekeeping genes are analyzed from the same sample as the target genes
• Detection directly from cell lysates: no need for RNA extraction or cDNA conversion

The transcripts detected by the CYP TRAC service include

Cytochrome P450 family enzymes
• Cyp1A1, Cyp1A2, Cyp2A6, Cyp2B6, Cyp2C8, Cyp2C9, Cyp2C19, Cyp2D6, Cyp2E1, Cyp3A4, CYP11A, CYP11B2, CYP17 and CYP19

Other genes:
• 17bHSD1, 17bHSD4, 3bHSD2, Por, Sult2A1, Ugt1A1, HMOX1, ABCA, ABCC2

Expression profiles of CYP3A4, CYP1A2 and CYP2B6 analyzed by TRAC in differently treated primary
hepatocytes from 3 donors. Error bars represent the standard error of the mean (SEM). Expression levels are
presented as fold change to vehicle treatment (DMSO). Data by Plexpress.

Promotional offer for CYP induction by TRAC

We offer a special 25% discount on CYP induction by TRAC studies, which have been ordered during February-March, 2012. The studies are conducted either by using cryopreserved primary hepatocytes or HepaRG cells. The detected transcripts include those listed above. Further customization to the study protocols are available, like used cell systems and detected transcripts.

The promotional offer also includes 25% discount on CYP inhibition and CYP identification studies, when ordered together with CYP TRAC.

Please contact us for more information and case studies comparing TRAC to other methods.

Adjunct professor and Research group leader, Dr. Moshe Finel has joined SBW Scientific Advisory Board.

Dr Finel is a Group leader at the Centre for Drug Research (CDR), Faculty of Pharmacy, University of Helsinki, Finland. Dr. Finel got his M.Sc. in Life Sciences in 1983 from the Weizmann Institute of Science, Rehovot, Israel and his Ph.D. in Biochemistry in 1989 from the University of Helsinki. He obtained postdoctoral training in the Laboratory of Molecular Biology, Medical Research Council, Cambridge, UK.

Dr. Finel’s research interests are in Phase II / conjugation drug metabolism, mainly the glucuronidation of drugs and related compounds, as well as the enzymes that catalyze these reactions, the UDP-glucuronosyltransferases (UGTs). Dr. Finel has published over 100 publications and he has supervised many Ph.D. theses.

>>Find our more on Adjunct professor Finel’s research on this page

Pharmacokinetic studies can become a bottleneck in the screening phase due to the high number of samples collected to assess the PK properties of multiple molecules, in various time points, and by alternative administration routes. Cassette dosing, simultaneous administration of multiple molecules in one animal, dramatically reduces the number of samples collected and animals sacrificed, and consequently, saves time and money. Read more on our PK expertise from this link.

In December 2011 we offer SimaPilot -a cassette dosing study:
* Up to 5 simultaneously administered molecules
* In rat or in mouse
* With 5 time points, 2 animals per time point
* With iv and po administration
* The parent concentrations are analysed by UPLC/MS-MS

During the campaign, the price of the SimaPilot includes free Plasma Protein Binding (RED) for the same molecules in rat or mouse.

This offer is valid until the end of 2011. The studies will be conducted in January – February, 2012. Maximum of one cassette of compounds per a customer.

Please send price list and quotation on request to info@sbw.fi or connect to us from this page.

We have a new in vivo efficacy model available for Inflammatory bowel disease, like ulcerative colitis and Crohn disease mice model. This is a mice model, where colitis is induced with dextran sodium sulfate (DSS).

“Crohn’s disease causes inflammation of the digestive system. It is one of a group of diseases called inflammatory bowel disease. The disease can affect any area from the mouth to the anus. It often affects the lower part of the small intestine called the ileum.”National Institute of Diabetes and Digestive and Kidney Diseases

The acute DSS colitis model in susceptible mice strains is particularly useful to study the contribution of innate immune mechanisms of colitis.

The Average Colitis Index (ACI) is calculated, including the body weight, faeces consistency and faecal blood. Experimental groups were compared to healthy controls that were access to normal tap water. The results were evaluated statistically.

See more on our Gastro-Intestinal models on this page and an overview of our in vivo disease models on this link.