Automated Applications of Sandwich-Cultured Hepatocytes in the Evaluation of Hepatic Drug Transport

Predictions of the absorption, distribution, metabolism, excretion, and toxicity of compounds in pharmaceutical development are essential aspects of the drug discovery process. B-CLEAR is an in vitro system that uses sandwich-cultured hepatocytes to evaluate and predict in vivo hepatobiliary disposition (hepatic uptake, biliary excretion, and biliary clearance), transporter-based hepatic drug-drug interactions, and potential drug-induced hepatotoxicity. Automation of predictive technologies is an advantageous and preferred format in drug discovery. In this study, manual and automated studies are investigated and equivalence is demonstrated. In addition, automated applications using model probe substrates and inhibitors to assess the cholestatic potential of drugs and evaluate hepatic drug transport are examined. The successful automation of this technology provides a more reproducible and less labor-intensive approach, reducing potential operator error in complex studies and facilitating technology transfer.

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A NEW GUIDANCE FOR THE PREDICTION OF HEPATIC CLEARANCE IN THE EARLY DRUG DISCOVERY AND DEVELOPMENT FROM THE IN VITRO-TO-IN VIVO EXTRAPOLATION METHOD AND AN APPROACH FOR EXPLORING WHETHER AN ALBUMIN-MEDIATED HEPATIC UPTAKE PHENOMENON COULD BE PRESENT UNDER IN VIVO CONDITIONS

Journal of Pharmaceutical Sciences ◽

10.1016/j.xphs.2021.04.002 ◽

2021 ◽

Author(s):

Patrick Poulin ◽

Sami Haddad

Keyword(s):

Drug Discovery ◽

Hepatic Clearance ◽

Hepatic Uptake ◽

Extrapolation Method ◽

Drug Discovery And Development ◽

In Vivo Extrapolation ◽

Early Drug

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Current Approaches to Drug Discovery for Chagas Disease: Methodological Advances

Combinatorial Chemistry & High Throughput Screening ◽

10.2174/1386207322666191010144111 ◽

2019 ◽

Vol 22 (8) ◽

pp. 509-520

Author(s):

Cauê B. Scarim ◽

Chung M. Chin

Keyword(s):

Drug Discovery ◽

Chagas Disease ◽

Drug Candidates ◽

Lead Compounds ◽

New Drug ◽

Compound Libraries ◽

Screening Assays ◽

Cruzi Infection

Background: In recent years, there has been an improvement in the in vitro and in vivo methodology for the screening of anti-chagasic compounds. Millions of compounds can now have their activity evaluated (in large compound libraries) by means of high throughput in vitro screening assays. Objective: Current approaches to drug discovery for Chagas disease. Method: This review article examines the contribution of these methodological advances in medicinal chemistry in the last four years, focusing on Trypanosoma cruzi infection, obtained from the PubMed, Web of Science, and Scopus databases. Results: Here, we have shown that the promise is increasing each year for more lead compounds for the development of a new drug against Chagas disease. Conclusion: There is increased optimism among those working with the objective to find new drug candidates for optimal treatments against Chagas disease.

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Assessing Drug Transport Across the Human Placental Barrier: From In Vivo and In Vitro Measurements to the Ex Vivo Perfusion Method and In silico Techniques

Current Pharmaceutical Biotechnology ◽

10.2174/138920111795470930 ◽

2011 ◽

Vol 12 (5) ◽

pp. 804-813 ◽

Cited By ~ 11

Author(s):

Constantinos Giaginis ◽

Anna Tsantili-Kakoulidou ◽

Stamatios Theocharis

Keyword(s):

In Silico ◽

Drug Transport ◽

Ex Vivo ◽

Placental Barrier ◽

Ex Vivo Perfusion ◽

Perfusion Method ◽

In Vitro Measurements

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Enhancement of hepatic 4-hydroxylation of 25-hydroxyvitamin D3through CYP3A4 induction in vitro and in vivo: Implications for drug-induced osteomalacia

Journal of Bone and Mineral Research ◽

10.1002/jbmr.1839 ◽

2013 ◽

Vol 28 (5) ◽

pp. 1101-1116 ◽

Cited By ~ 38

Author(s):

Zhican Wang ◽

Yvonne S Lin ◽

Leslie J Dickmann ◽

Emma-Jane Poulton ◽

David L Eaton ◽

...

Keyword(s):

Drug Induced ◽

Cyp3a4 Induction

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Consideration of albumin-mediated hepatic uptake for highly protein-bound anionic drugs: Bridging the gap of hepatic uptake clearance between in vitro and in vivo

Pharmacology & Therapeutics ◽

10.1016/j.pharmthera.2021.107938 ◽

2021 ◽

pp. 107938

Author(s):

Seiji Miyauchi ◽

Soo-Jin Kim ◽

Wooin Lee ◽

Yuichi Sugiyama

Keyword(s):

Hepatic Uptake ◽

Anionic Drugs

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Imidazolidinyl urea activates mast cells via MRGPRX2 to induce non-histaminergic allergy

Toxicology Research ◽

10.1093/toxres/tfab035 ◽

2021 ◽

Author(s):

Jiapan Gao ◽

Delu Che ◽

Xueshan Du ◽

Yi Zheng ◽

Huiling Jing ◽

...

Keyword(s):

Contact Dermatitis ◽

Mast Cells ◽

Pharmaceutical Products ◽

Drug Induced ◽

Inflammatory Infiltration ◽

Local Inflammatory Response ◽

Allergic Contact ◽

G Protein Coupled

Abstract Imidazolidinyl urea (IU) is used as an antimicrobial preservative in cosmetic and pharmaceutical products. IU induces allergic contact dermatitis, however, the mechanism has not yet been elucidated. Mas-related G protein-coupled receptor-X2 (MRGPRX2) triggers drug-induced pseudo-allergic reactions. The aims of this study were to determine whether IU activated mast cells through MRGPRX2 to further trigger contact dermatitis. Wild-type (WT) and KitW-sh/HNihrJaeBsmJNju (MUT) mice were treated with IU to observe its effects on local inflammation and mast cells degranulation in vivo. Laboratory of allergic disease 2 cells were used to detect calcium mobilization and release of inflammatory mediators in vitro. WT mice showed a severe local inflammatory response and contact dermatitis, whereas only slight inflammatory infiltration was observed in MUT mice. Thus, MRGPRX2 mediated the IU-induced activation of mast cells. However, histamine, a typical allergen, was not involved in this process. Tryptase expressed by mast cells was the major non-histaminergic inflammatory mediator of contact dermatitis. IU induced anaphylactic reaction via MRGPRX2 and further triggering non-histaminergic contact dermatitis, which explained why antihistamines are clinically ineffective against some chronic dermatitis.

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Expression of laminin γ 1 cultured hepatocytes involves repeated CTC and GC elements in the LAMC1 promoter

Biochemical Journal ◽

10.1042/bj3130745 ◽

1996 ◽

Vol 313 (3) ◽

pp. 745-752 ◽

Cited By ~ 5

Author(s):

Françoise LEVAVASSEUR ◽

Jocelyne LIÉTARD ◽

Kohei OGAWA ◽

Nathalie THÉRET ◽

Peter D. BURBELO ◽

...

Keyword(s):

Transcriptional Activation ◽

Hepatoma Cell ◽

Primary Cultures ◽

Regulatory Elements ◽

Hepatoma Cells ◽

Basement Membranes ◽

Major Protein ◽

Cultured Hepatocytes

Laminin γ1 chain is present in all basement membranes and is expressed at high levels in various diseases, such as hepatic fibrosis. We have identified cis- and trans-acting elements involved in the regulation of this gene in normal rat liver, as well as in hepatocyte primary cultures and hepatoma cell lines. Northern-blot analyses showed that laminin γ1 mRNA was barely detectable in freshly isolated hepatocytes and expressed at high levels in hepatocyte primary cultures, as early as 4 h after liver dissociation. Actinomycin D and cycloheximide treatment in vivo and in vitro indicated that laminin γ1 overexpression in cultured hepatocytes was under the control of transcriptional mechanisms. Transfection of deletion mutants of the 5´ flanking region of murine LAMC1 gene in hepatoma cells that constitutively express laminin γ1 indicated that regulatory elements were located between -594 bp and -94 bp. This segment included GC- and CTC-containing motifs. Gel-shift analyses showed that two complexes were resolved with different affinity for the CTC sequence depending on the location of the GC box. The pattern of complex formation with nuclear factors from freshly isolated and cultured hepatocytes was different from that obtained with total liver and similar to that with hepatoma cells. Southwestern analysis indicated that several polypeptides bound the CTC-rich sequence. Affinity chromatography demonstrated that a Mr 60000 polypeptide was a major protein binding to the CTC motif. This polypeptide is probably involved in the transcriptional activation of various proto-oncogenes and extracellular matrix genes that are expressed at high levels in both hepatoma cells and early hepatocyte cultures.

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In Vivo and In Vitro Toxicodynamic Analyses of New Quinolone-and Nonsteroidal Anti-Inflammatory Drug-Induced Effects on the Central Nervous System

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.43.5.1091 ◽

1999 ◽

Vol 43 (5) ◽

pp. 1091-1097 ◽

Cited By ~ 7

Author(s):

Hideki Kita ◽

Hirotami Matsuo ◽

Hitomi Takanaga ◽

Junichi Kawakami ◽

Koujirou Yamamoto ◽

...

Keyword(s):

Brain Tissue ◽

Threshold Concentration ◽

Current Response ◽

Drug Induced ◽

Inhibition Ratio ◽

New Quinolones ◽

The Central Nervous System ◽

Anti Inflammatory

ABSTRACT We investigated the correlation between an in vivo isobologram based on the concentrations of new quinolones (NQs) in brain tissue and the administration of nonsteroidal anti-inflammatory drugs (NSAIDs) for the occurrence of convulsions in mice and an in vitro isobologram based on the concentrations of both drugs for changes in the γ-aminobutyric acid (GABA)-induced current response in Xenopus oocytes injected with mRNA from mouse brains in the presence of NQs and/or NSAIDs. After the administration of enoxacin (ENX) in the presence or absence of felbinac (FLB), ketoprofen (KTP), or flurbiprofen (FRP), a synergistic effect was observed in the isobologram based on the threshold concentration in brain tissue between mice with convulsions and those without convulsions. The three NSAIDs did not affect the pharmacokinetic behavior of ENX in the brain. However, the ENX-induced inhibition of the GABA response in the GABAA receptor expressed in Xenopus oocytes was enhanced in the presence of the three NSAIDs. The inhibition ratio profiles of the GABA responses for both drugs were analyzed with a newly developed toxicodynamic model. The inhibitory profiles for ENX in the presence of NSAIDs followed the order KTP (1.2 μM) > FRP (0.3 μM) > FLB (0.2 μM). These were 50- to 280-fold smaller than those observed in the absence of NSAIDs. The inhibition ratio (0.01 to 0.02) of the GABAA receptor in the presence of both drugs was well-fitted to the isobologram based on threshold concentrations of both drugs in brain tissue between mice with convulsions and those without convulsions, despite the presence of NSAIDs. In mice with convulsions, the inhibitory profiles of the threshold concentrations of both drugs in brain tissue of mice with convulsions and those without convulsions can be predicted quantitatively by using in vitro GABA response data and toxicodynamic model.

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