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We offer an extended range of non-clinical services to the customers requiring reliable data for lead optimization prior to in vivo testing. We help to obtain the data necessary for the effective and well-grounded decision-making using our cost-effective high-throughput in vitro Adsorption, Distribution, Metabolism, and Excretion (In Vitro ADME) testing of the compounds. In Vitro ADME is an essential step in the drug development process, which may critically influence the success of the entire drug development program. The use of our in vitro high-throughput ADME screening protocols affords a significant saving of time, labor, and other resources. Ask about our unique assay miniaturization to save your precious testing material.

We focus on the most critical but also high-quality labor-intensive procedures comprising the In Vitro ADME, such as metabolism and permeability evaluation.

Cell Proliferation Assay

In our 384-well high throughput assay, we are using high content imaging to visualize and count cells after exposure to chemicals. The cells are treated with a wide range of concentrations of the tested compound and at the end of the exposure, the cells are labeled with live-dead viability dyes, and the plate is imaged with INCell 2200 high content imager. Data is analyzed for  The dose-response curves are plotted and analyzed. The experiment is repeated three times on three independent days. 

The high throughput format allows us to screen many compounds at the same time.

Since we are using high content imaging for our assays, additional biomarkers can be included in the assay and evaluated at the same.

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Hepatocyte Stability Assay

The liver is a guardian of human health. Liver enzymes eliminate half of the marketed drugs producing a pool of metabolites specific to each drug. Chemicals and drug metabolites can be toxic to the body, therefore, it is essential to know of the clearance rate of a drug candidate and both the identity and the clearance rate of its metabolites.

We offer hepatocyte stability assay, which involves a drug incubation with live hepatocytes using primary hepatocytes or immortalized cell lines, such as HepRG cells – cells containing phase I and phase II metabolizing enzymes. Cells and cell culture media are used to quantify the amount of drug and to identify the metabolites. This assay allows to determine the fate of the drug in the body and calculate its clearance rate. Using species-specific hepatocytes, we can help to determine the species-specificity of hepatic clearance and translate animal data to the human situation.

Microsome Stability Assay

Human microsomes provide convenience and high reproducibility of an acellular assay. Hepatic microsomes contain CYP450 enzymes that normally reside in the endoplasmic reticulum. While not providing a full spectrum of liver metabolizing enzymes, microsomal stability assay is less prone to variability associated with donor-specific nature of human primary hepatocytes and is a cost-saving alternative. This metabolic data from the assay can then be used to pinpoint any metabolic liabilities at an early stage to help improve drug candidates. In addition to hepatic microsomes, we offer stability and metabolism assay using microsomes derived from skin, intestine, and endothelium (as a model for in blood-brain barrier).

Plasma Stability Assay

A plasma stability assay, as part of our in vitro ADME assay screenings, can be used to asses how much your compounds will degrade once it enters the bloodstream. It can be combined with drug absorption measurement to determine the percent of an active/unbound drug in the blood. This can be particularly important for antedrugs and prodrugs screenings, where rapid conversion is particularly desirable. Generally, drugs that degrade quickly also have poor in vivo efficacy. If your compound has a functional group such as amides, lactams, esters, sulphonamides, lactones, or carbamides, then they may be more likely to succumb to hydrolysis in plasma. The actual plasma instability of a drug can significantly affect decision-making after initial in vitro screening results. It’s also possible for inter-species differences for plasma degradation and species-specific binding of testing compounds, which reduces the efficacy of a drug. Therefore, we offer both human and animal plasma testing.

Metabolite Profiling and Identification

Discovering any metabolites formed is an important step in drug discovery, as it brings awareness to potentially toxic metabolites that can form as well as leading the way for a more metabolically stable drug. Thus, to further understand the fate of your compound in the human body, you may want to go on and identify the metabolites that are formed during incubation with cells, subcellular organelles, or plasma. As part of our In Vitro ADME service, we will identify these metabolites. Using our data, you will be able to predict the pharmacological and toxicological consequences of exposure to your drug candidate.

Permeability Assay

As part of our In Vitro ADME screening, the Caco-2 permeability assay will help to determine oral bioavailability of a compound as well as its safety.  In this assay, we will determine the compound’s rate of transport through the barrier formed by the Caco-2 cell line. As a model for the intestine barrier, Caco-2 cell line is a golden standard for in vitro permeability testing characterized by specific tight junctions between cells that form a polarized monolayer. Our assay can assess the transport of the compound in both directions to provide an indication of whether or not the drug undergoes active efflux.

Another highly popular model for intra-body barriers is the MDCK cell line, which is used as an effective surrogate for kidney and gastrointestinal epithelia and even as a blood-brain barrier.  

We also offer a 96-well high-throughput parallel artificial membrane permeability assay (PAMPA). This assay provides exceptional reproducibility because it does not involve live cells but still can provide essential information about a passive permeability of a drug, which is especially beneficial for conditions involving a range of pH values – from the acidic environment of the stomach to the alkaline environment of the small intestine. This will give an understanding of how a drug is taken orally will be absorbed in different parts of the gastrointestinal tract.

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