Validation of Human MDR1-MDCK and BCRP-MDCK Cell Lines to Improve the Prediction of Brain Penetration

Feng, Bo; West, Mark; Patel, Nandini; Wager, Travis T.; Hou, Xinjun; Johnson, Jillian G.; Tremaine, Larry M.; Liras, Jennifer

doi:10.1016/j.xphs.2019.02.005

Cited by 56 publications

(75 citation statements)

References 18 publications

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“…Bidirectional permeability assay using MDR1-transfected MDCK cells is a well-accepted in vitro screening assay in the pharmaceutical industry to address the blood–brain barrier efflux [12,13,21]. However, since the expression levels of the transporters in the cell lines used in different labs may differ strongly [21], it is important for each lab to calibrate its in vitro screening assay using in vivo data. In our case we compared the in vitro efflux measured in our in-house MDR1-overexpressing MDCK cells (MDCK-MDR1) with K p,br/mu measured in rats.…”

Section: Resultsmentioning

confidence: 99%

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Muscle to Brain Partitioning as Measure of Transporter-Mediated Efflux at the Rat Blood–Brain Barrier and Its Implementation into Compound Optimization in Drug Discovery

et al. 2019

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Movement of xenobiotic substances across the blood–brain barrier (BBB) is tightly regulated by various transporter proteins, especially the efflux transporters P-glycoprotein (P-gp/MDR1) and breast cancer resistance protein (BCRP). Avoiding drug efflux at the BBB is a unique challenge for the development of new central nervous system (CNS) drugs. Drug efflux at the BBB is described by the partition coefficient of unbound drug between brain and plasma (Kp,uu,brain) which is typically obtained from in vivo and often additionally in vitro measurements. Here, we describe a new method for the rapid estimation of the in vivo drug efflux at the BBB of rats: the measurement of the partition coefficient of a drug between brain and skeletal muscle (Kp,brain/muscle). Assuming a closely similar distribution of drugs into the brain and muscle and that the efflux transporters are only expressed in the brain, Kp,brain/muscle, similar to Kp,uu,brain, reflects the efflux at the BBB. The new method requires a single in vivo experiment. For 64 compounds from different research programs, we show the comparability to other approaches used to obtain Kp,uu,brain. P-gp- and BCRP-overexpressing cell systems are valuable in vitro tools for prescreening. Drug efflux at the BBB can be most accurately predicted based on a simple algorithm incorporating data from both in vitro assays. In conclusion, the combined use of our new in vivo method and the in vitro tools allows an efficient screening method in drug discovery with respect to efflux at the BBB.

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Section: Resultsmentioning

confidence: 99%

“…Bidirectional permeability measurement using transfected MDCK cells is a well-accepted in vitro screening assay in the pharmaceutical industry to address the blood–brain barrier efflux [12,13,21]. We describe in this work the calibration of our in-house transfected cell lines with the in vivo K p,br/mu data.…”

Section: Discussionmentioning

confidence: 99%

Muscle to Brain Partitioning as Measure of Transporter-Mediated Efflux at the Rat Blood–Brain Barrier and Its Implementation into Compound Optimization in Drug Discovery

et al. 2019

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“…The blood-brain barrier (BBB) is considered as the bottleneck in brain drug development and is an important consideration in determining whether a drug can penetrate the brain during treatment of glioblastoma [39]. Multidrug resistance protein 1 (MDR1) and breast cancer resistance protein (BCRP) are two important efflux drug transporters that are highly expressed at the BBB [40]. These can actively expel these substrates from the brain, limiting the penetration of compounds in the brain [40].…”

Section: Otssp167 Inhibits the Growth Of Gbm Xenografts In Vivomentioning

confidence: 99%

“…Multidrug resistance protein 1 (MDR1) and breast cancer resistance protein (BCRP) are two important efflux drug transporters that are highly expressed at the BBB [40]. These can actively expel these substrates from the brain, limiting the penetration of compounds in the brain [40]. A recent study has shown that OTSSP167 is a substrate of MDR1 and BCPR.…”

Section: Otssp167 Inhibits the Growth Of Gbm Xenografts In Vivomentioning

confidence: 99%

MELK inhibition effectively suppresses growth of glioblastoma and cancer stem-like cells by blocking AKT and FOXM1 pathways

Liu

Wang

et al. 2020

Preprint

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Background: Glioblastoma multiforme (GBM) is a devastating disease yet no effective drug treatment has been established to date. Glioblastoma stem-like cells (GSCs) are insensitive to treatment and may be one of the reasons for the relapse of GBM. Maternal embryonic leucine zipper kinase gene (MELK) plays an important role in the malignant proliferation and the maintenance of GSC stemness properties of GBM. However, the therapeutic effect of targeted inhibition of MELK on GBM remains unclear. Methods: This study analyzed the effect of a MELK oral inhibitor, OTSSP167, on GBM proliferation and the maintenance of GSC stemness. Results: OTSSP167 significantly inhibited cell proliferation, colony formation, invasion, and migration of GBM. OTSSP167 treatment reduced the expression of cell cycle G2/M phase-related proteins, Cyclin B1 and Cdc2, while up-regulation the expression of cell cycle inhibitor protein p21 and subsequently induced cell cycle arrest at the G2/M phase. OTSSP167 effectively prolonged the survival of tumor-bearing mice and inhibited tumor cell growth in an in vivo mouse model. It also reduced protein kinase B (Akt) phosphorylation by inhibiting MELK, thereby disrupting the proliferation and invasion of GBM cells. Furthermore, OTSSP167 inhibited the proliferation and neurosphere formation of GSCs by reducing forkhead box M1 (FOXM1) phosphorylation and transcriptional activity. Interestingly, the inhibitory effect of OTSSP167 on the proliferation of GSCs was 4-fold more effective than GBM cells. Conclusion: In conclusion, MELK inhibition suppresses the growth of GBM and GSCs by double-blocking AKT and FOXM1 signals. Targeted inhibition of MELK may thus be potentially used as a novel treatment for GBM.

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“…4 Nevertheless, the MDCK-MDR1-transfected cell line has become a popular tool similar to the Caco2 model and is used as an in vitro marker of brain transport of P-gp substrates due to the very tight junctions available on this canine model. 5 These are the only two cell lines that currently provide tight junctions adequate enough for bidirectional drug transport where evidence of efflux can be recorded, usually expressed as an efflux ratio of drug moved out to drug moving into the cells that enables researchers to define the potential of a drug to be either a substrate or evidence of potential inhibitors' effectiveness.…”

Section: Introductionmentioning

confidence: 99%

P-Glycoprotein-Mediated Efflux Using a Rapidly Maturing Caco2 Clone (CLEFF4) in Only 5 Days without Requiring Modified Growth Medium

Crowe

2021

SLAS Discovery

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Validation of Human MDR1-MDCK and BCRP-MDCK Cell Lines to Improve the Prediction of Brain Penetration

Cited by 56 publications

References 18 publications

Muscle to Brain Partitioning as Measure of Transporter-Mediated Efflux at the Rat Blood–Brain Barrier and Its Implementation into Compound Optimization in Drug Discovery

Muscle to Brain Partitioning as Measure of Transporter-Mediated Efflux at the Rat Blood–Brain Barrier and Its Implementation into Compound Optimization in Drug Discovery

MELK inhibition effectively suppresses growth of glioblastoma and cancer stem-like cells by blocking AKT and FOXM1 pathways

P-Glycoprotein-Mediated Efflux Using a Rapidly Maturing Caco2 Clone (CLEFF4) in Only 5 Days without Requiring Modified Growth Medium

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