In vitro assessment of P450 induction potential of novel chemopreventive agents SR13668, 9-cis-UAB30, and pentamethychromanol in primary cultures of human hepatocytes

Jackson, Jonathan P.; Kabirov, Kasim K.; Kapetanović, Izet M.; Lyubimov, Alexander V.

doi:10.1016/j.cbi.2008.12.005

Cited by 13 publications

(15 citation statements)

References 19 publications

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“…The high binding affinity of human PXR for rifampicin followed by PCN has been observed by other researchers [7,9,15,28,40,41]. Although, 3-MC is predicted to have a low binding energy to the LBD of human PXR [42], it has been shown to be activated by members of the cytochrome P450 family of enzymes, which include activating CYP1A1/2 enzymes [42][43][44][45]. The magnitude response order of rifampicin > PCN > 3-MC is what we predicted from the literature.…”

Section: Detection Of Known Pxr Activators Andsupporting

confidence: 57%

Microcantilever-based Nanomechanical Studies of the Orphan Nuclear Receptor Pregnane X Receptor-Ligand Interactions

Hill¹,

Dutta²,

Zhang³

2011

JBNB

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Human pregnane X receptor (PXR) is of vital importance in pharmaceutical and exogenous compound metabolism within the body. This provides strong motivation for investigating this orphan receptor's activation by various pharmaceuticals, xenobiotics, and endocrine disrupting chemicals (EDCs). A nanomechanical transducer is developed to study xenobiotic and EDC interactions with the bioreceptor PXR's ligand binding domain (LBD). The combination of immobilized LBD PXR with a nanostructured microcantilever (MC) platform allows for the sensitive, label-free study of ligand interaction with the receptor. PXR shows real-time, reversible responses when exposed to a specific pharmaceutical, EDCs, and xenobiotic ligands. Three EDCs binding interactions are tested, which include phthalic acid, nonylphenol, and bisphenol A, with PXR. PXR LBD was exposed to rifampicin, a potent PXR activator, with various injection and recovery times to study their interaction. A two protein array of PXR and estrogen receptor  (ER-) directlycompares the nanomechanical responses of these receptors with rifampicin on a single platform.

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Section: Detection Of Known Pxr Activators Andsupporting

confidence: 57%

Microcantilever-based Nanomechanical Studies of the Orphan Nuclear Receptor Pregnane X Receptor-Ligand Interactions

Hill¹,

Dutta²,

Zhang³

2011

JBNB

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“…The following final substrate concentrations were used in cell culture media: 100 mM phenacetin (CYP1A2), 5 mM coumarin (CYP2A6), 500 mM bupropion (CYP2B6), 20 mM paclitaxel (CYP2C8), 25 mM diclofenac (CYP2C9), 250 mM S-mephenytoin (CYP2C19), 15 mM dextromethorphan (CYP2D6), 250 mM chlorzoxazone, 10 mM midazolam (CYP3A4/5), 200 mM testosterone (CYP3A4/5), 250 mM benzydamine (FMO), or 100 mM 7-hydroxycoumarin (sulfotransferase, UDPglucuronosyltransferase). All in situ metabolism assays were conducted in singleprobe format as previously described (Jackson et al, 2009). All cell suspension metabolism assays were performed in single probe format as previously described (Smith et al, 2012).…”

Section: Methodsmentioning

confidence: 99%

“…Metabolism assay samples were collected and stored frozen at -80°C until they were processed for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. LC-MS/MS analysis of metabolism assay samples was conducted as previously described (Jackson et al, 2009;Smith et al, 2012) and included standard curves with at least six calibration standards along with 12 quality control samples (at three different concentrations) dispersed at the beginning, middle, and end of analytical runs to assess quantitative continuity throughout the run.…”

Section: Methodsmentioning

confidence: 99%

Contextualizing Hepatocyte Functionality of Cryopreserved HepaRG Cell Cultures

Jackson

Chamberlain

et al. 2016

Drug Metabolism and Disposition

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Over the last decade HepaRG cells have emerged as a promising alternative to primary human hepatocytes (PHH) and have been featured in over 300 research publications. Most of these reports employed freshly differentiated HepaRG cells that require timeconsuming culture (∼28 days) for full differentiation. Recently, a cryopreserved, predifferentiated format of HepaRG cells (termed here "cryo-HepaRG") has emerged as a new model that improves global availability and experimental flexibility; however, it is largely unknown whether HepaRG cells in this format fully retain their hepatic characteristics. Therefore, we systematically investigated the hepatocyte functionality of cryo-HepaRG cultures in context with the range of interindividual variation observed with PHH in both sandwich-culture and suspension formats. These evaluations uncovered a novel adaptation period for the cryo-HepaRG format and demonstrated the impact of extracellular matrix on cryo-HepaRG functionality. Pharmacologically important drug-metabolizing alleles were genotyped in HepaRG cells and poor metabolizer alleles for CYP2D6, CYP2C9, and CYP3A5 were identified and consistent with higher frequency alleles found in individuals of Caucasian decent. We observed liver enzyme inducibility with aryl hydrocarbon receptor, constitutive androstane receptor (CAR), and pregnane X receptor activators comparable to that of sandwich-cultured PHH. Finally, we show for the first time that cryo-HepaRG supports proper CAR cytosolic sequestration and translocation to hepatocyte nuclei in response to phenobarbital treatment. Taken together, these data reveal important considerations for the use of this cell model and demonstrate that cryo-HepaRG are suitable for metabolism and toxicology screening.

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“…In vitro studies with human liver microsomes (HLMs) suggest that 9-cis-UAB30 has low potential to induce CYP1A2 and CYP3A4 expression at the concentrations examined. However, 9-cis-UAB30 was shown to significantly induce CYP2B6 enzyme activity at 10 μM (Jackson et al, 2009) Hypervitaminosis A results in liver abnormalities, including fibrosis and cirrhosis (Roenigk, 1988). Retinoids have been observed to cause hepatic changes, due to vitamin A metabolism involving storage in the liver.…”

Section: Research Articlementioning

confidence: 97%

Assessment of oral toxicity and safety of 9-cis-UAB30, a potential chemopreventive agent, in rat and dog studies

Lindeblad

Kapetanović

Kabirov

et al. 2011

Drug and Chemical Toxicology

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9-cis-UAB30 is a potential chemopreventative agent that has been shown to be effective on many different types of tumors. The safety and toxicity of 9-cis-UAB30 had not been previously established. These studies were conducted to evaluate the potential toxicity and pharmacokinetics in a rodent and a nonrodent species for the purpose of investigational new drug submission. Oral gavage administration of 9-cis-UAB30 at the doses 0, 3, 15, and 100 mg/kg/day to CD® rats for 28 days showed a dose-dependent (although not dose-proportional) increase in plasma drug levels in week 4. The liver was the target organ for toxicity of 9-cis-UAB30. Hepatomegaly along with increases in serum aspartate-aminotransferase and alkaline-phosphatase levels were seen in rats. Moderate hypoalbuminemia and hyperglobulinemia resulted in a decreased albumin/globulin ratio. Histopathology revealed hepatocellular change consistent with hepatic glycogen deposition. Toxicity studies in dogs did not show treatment-related toxicity at doses as high as 100 mg/kg/day (highest dose tested) administered by capsules for 28 days. No effects on the central nervous system (functional observational battery in rats) or cardiovascular function (safety pharmacology study in telemeterized dogs) were seen. The no observed adverse effect level (NOAEL) in the rat studies was 3 mg/kg/day; however, the adverse effects seen in rats receiving 15 mg/kg/day (the least observed adverse effect level) was a slight, but statistically significant, elevation in fibrinogen and decrease in prothrombin time, which may be a sign of some tendency for increased blood coagulation. The NOAEL in the dog study was at least 100 mg/kg/day.

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In vitro assessment of P450 induction potential of novel chemopreventive agents SR13668, 9-cis-UAB30, and pentamethychromanol in primary cultures of human hepatocytes

Cited by 13 publications

References 19 publications

Microcantilever-based Nanomechanical Studies of the Orphan Nuclear Receptor Pregnane X Receptor-Ligand Interactions

Microcantilever-based Nanomechanical Studies of the Orphan Nuclear Receptor Pregnane X Receptor-Ligand Interactions

Contextualizing Hepatocyte Functionality of Cryopreserved HepaRG Cell Cultures

Assessment of oral toxicity and safety of 9-cis-UAB30, a potential chemopreventive agent, in rat and dog studies

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