Toxicology Assay Kits

Rapidly assess therapeutic candidates’ potential for drug-drug interactions and accurately predict xenobiotic-induced liver toxicity. INDIGO’s toxicology assay kits provide rapid, reliable results for:

Prospective screening

Retrospective screening

Safety pharmacology

Showing all 3 results

Expression Profiling of Clinically Relevant CYPs Assay Kit

Species

Human

This Expression Profiling of Clinically Relevant CYPs assay kit contains optimized reagents for the culturing and treatment of upcyte® hepatocytes to assess drug-induced changes in CYP3A4, CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP2E1.

The kit provides two aliquots of upcyte® hepatocytes, two cell culture-ready assay plates, optimized Cell Culture Medium for use in all steps of the assay procedure, and three reference compounds (rifampicin, β-naphthoflavone, and CDCA) that activate one or more of the primary xenobiotic-sensing receptors: PXR, CAR, AhR, and FXR. Upon activation, these nuclear receptors modulate the expression of the CYP genes. Also included are seven sets of validated qPCR primers for quantifying drug-induced changes in the expression of CYP3A4, CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP2E1, as well as primers for ACTB.

Please note: This kit does not include reagents or protocols for cell lysis, RNA isolation, cDNA preparation, or qPCR assays.

Human P-Glycoprotein / MDR1 Drug Interaction Assay

Species

Human

The primary application of INDIGO Biosciences MDR1 drug interaction assay kit is to rapidly assess drug candidates as either inhibitors, substrates, or non-substrates of P-glycoprotein. This is accomplished by co-treating the cells with varying concentrations of the test drug and a fixed concentration of daunorubicin, a well-characterized fluorescent substrate for P-glycoprotein mediated transport. HCT-Pgp cells are a proprietary cell line that have been extensively selected for high-level expression of native P-glycoprotein.

The kit contains two aliquots of HCT-Pgp cells and two black, sterile, collagen-coated 96-well assay plates. In addition to HCT-Pgp cells and assay plates, this kit includes Cell Recovery Medium (CRM-p) used to perform a rapid thaw of the -80°C HCT-Pgp cells, Compound Screening Medium supplemented with daunorubicin (CSM+DR) used to prepare drug treatment media, Wash Buffer, two aliquots of Lysis Reagent, and the potent reference inhibitor Tariquidar.

in vitro Screening for Drug-Induced Hepatotoxicity Assay Kit

Species

Human

The principal application of this assay is to rapidly screen test compounds to identify those that induce acute liver cell toxicity.

This kit includes two aliquots of cryopreserved Luminescent Reporter Hepatocytes (upcyte®), donor 10-13, isolated from an adult Caucasian female, that have been further modified to constitutively express the luciferase enzyme. In addition to two aliquots of the reporter hepatocytes, the kit provides two cell culture-ready assay plates, optimized Cell Culture Media (CCM) for use in all steps of the assay procedure (cell thawing, seeding, and preparation of the treatment media), luciferase detection reagent, and a reference compound that provides a positive control for hepatotoxicity.

The reagents and materials provided in this assay kit are formatted to allow the user to choose between two alternative assay setups. In one scenario 48 culture wells may be setup at two different times. In the other assay scenario 96 culture wells may be setup at one time.

upcycte Hepatocytes in INDIGO Toxicology Kits

Xenobiotic-induced liver injury is a major cause of human morbidity and mortality. A key reason for this problem is our inability to predict hepatotoxicity at the preclinical stage using currently available model systems. Such models include in vivo animal models and in vitro models based on human-derived liver cells or transformed cell systems. Species differences in xenobiotic disposition and mechanisms of cytotoxicity can make whole animal studies unreliable for extrapolation to humans. In addition, whole animal models are costly and of low throughput. Therefore, it is essential to develop in vitro models that are more predictive of hepatotoxicity, particularly those that are based on human or "humanized" component cells.

There are two major limitations to the use of human liver cells or their derivatives. First, there are currently limited sources of fresh human hepatocytes worldwide and, when available, they often suffer from low viability and high batch-to-batch variability. Second, the freezing process used to preserve primary hepatocytes and the transformation process needed to make stable and proliferating cell lines results in changes in cell differentiation, proliferation, and metabolic processes.

INDIGO's in vitro toxicology platform was developed to meet the increasing demand for more predictive models for liver toxicity due to the fact that hepatotoxicity remains a major reason for drug withdrawal from pharmaceutical development and clinical use. Consequently, the use of hepatocytes for the early identification of drug candidates that induce acute hepatotoxicity provides a powerful predictive tool that can inform drug development decisions.

Application of this platform is to screen chemicals for induction and activity of hepatic drug metabolizing enzymes (DME), which have been linked to Non-Alcoholic Fatty Liver Disease, Steatosis, Cholestasis and other conditions.

The platform now utilizes upcyte^® human liver cells to assess chemical and drug-induced toxicity.

EXPAND CONTENT

Expression Profiling of Clinically Relevant CYPs

Cytochrome p450 (CYP) enzymes are responsible for the Phase I metabolism of most drugs. And, it is noteworthy that the expression of cytochrome p450 (CYP) genes are predominantly regulated by ligand-activated receptors/transcription factors such as pregnane X receptor (PXR, NR1I2), constitutive androstane receptor (CAR, NR1I3), aryl hydrocarbon receptor (AhR), farnesoid x receptor (FXR, NR1H4), glucocorticoid receptor (GR, NR3C1), and to lesser degrees liver X receptors (LXR, NR1C3), vitamin D receptor (VDR, NR1I1), and peroxisome proliferator-activated receptor alpha (PPARα, NR1C1). Consequently, drugs that activate any of these xenobiotic sensing receptors can dramatically change the endogenous levels of CYP expression in the liver, potentially impacting the rate of their own metabolism, as well as the metabolism of all other co-administered drugs. Of particular concern are metabolic outcomes that transform a drug to greater potency, or to an altered bioactivity.

Assessing drug-induced changes in the expression of CYP genes provides a reliable predictive indicator of altered (either heightened or inhibited) metabolic activities leading to drug-drug interactions in vivo. Cytochrome p450 enzymes with the greatest clinical relevance belong to CYP1, 2, and 3 families. It is estimated that they are involved in the metabolism of 70-80% of drugs currently on the market. For example, CYP3A4 is estimated to be involved in ~30% of all prescription drugs.

INDIGO's assay kit for the Expression Profiling of Clinically Relevant CYPs contains optimized reagents for the culturing and treatment of upcyte^® hepatocytes to assess drug-induced changes in seven clinically-relevant CYPs: CYP3A4. CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP2E1.

EXPAND CONTENT

What is MDR1?

Human P-glycoprotein (P-gp) is also known as MDR1 or ABCB1, the human multidrug resistance protein 1. It is a 170kDa transmembrane glycoprotein that functions as an ATP-dependent efflux transporter present in the human body’s epithelial tissues.

P-gp consists of twelve membrane-spanning domains and two cytosolic nucleotide-binding domains. The transmembrane domains comprise several substrate binding pockets capable of interacting with a broad range of both endogenous and foreign small molecule chemotypes. Xenobiotic substrates of P-gp range from pollutants, such as those encountered through unintended exposure to industrial and agricultural chemicals, to small molecule drugs that are intentionally administered for therapeutic benefit.

P-gp is highly expressed in gastrointestinal epithelium, liver, pancreatic and kidney cells, and capillary endothelial cells that establish the blood-brain barrier. Multidrug resistance research has found MDR1 expressed at high levels on the lines of transformed and tumor cells. In combination with the activities of Cytochrome P450 oxidases, the robust efflux activity of P-gp plays a critical role in limiting the absorption and systemic physiological distribution of xenobiotics and facilitating their ultimate elimination from the body. The MDR1 transporter role is important for drug-drug interaction (DDI) and drug safety assessments.

EXPAND CONTENT

Toxicology Assay Kits

Expression Profiling of Clinically Relevant CYPs Assay Kit

Human P-Glycoprotein / MDR1 Drug Interaction Assay

in vitro Screening for Drug-Induced Hepatotoxicity Assay Kit

upcycte Hepatocytes in INDIGO Toxicology Kits

Expression Profiling of Clinically Relevant CYPs

What is MDR1?

Related Products and Services

Reporter Assay Services

Expression Profiling

Custom Assay Development