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Improved In Vitro-In Vivo Correlation by Using the Unbound-Fraction-Adjusted IC50 for Breast Cancer Resistance Protein Inhibition

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Abstract

Purpose

One function of the blood-brain barrier (BBB) is the efflux of xenobiotics by breast cancer resistance protein (BCRP), and inhibition of BCRP can cause unexpected central nervous system toxicity. Despite the importance of BCRP inhibition and the associated risk of BBB penetration in vivo, there has been little investigation of it to date. In this study, inhibition of BCRP-mediated transport was assessed by in vitro assay in the presence of bovine serum albumin (BSA) to change the unbound inhibitor concentrations, and the in vitro-in vivo correlation (IVIVC) at the BBB was evaluated.

Methods and Results

The IC50 values of BCRP inhibitors were determined in vitro with and without BSA and the inhibitors were categorized into two groups. One group of compounds had little risk of inhibiting BCRP because of their low unbound concentrations. In contrast, the other group has the potential to facilitate BBB penetration by inhibiting BCRP. In the IVIVC approach, brain concentrations and the brain-to-plasma ratio were better correlated with the ratio of the unbound plasma concentration at steady-state to the unbound-fraction-adjusted IC50.

Conclusion

We have found a way to obtain a better in vitro-in vivo correlation for BCRP-mediated transport.

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Abbreviations

BBB:

Blood-brain barrier

BSA:

Bovine serum albumin

BCRP (ABCG2):

Breast cancer resistance protein

Ct :

Total concentration

Cu :

Unbound concentration

DMSO:

Dimethyl sulfoxide

ER:

Efflux ratio

f u,1%BSA :

Unbound fraction with 1% BSA

f u,p :

Unbound fraction in plasma

HBSS:

Hanks’ Balanced Salt Solution

HEPES:

2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid

HPLC:

High-performance liquid chromatography

I ss,total :

Total plasma concentration at steady-state

I ss,unbound :

Unbound plasma concentration at steady-state

IVIVC:

In vitro-in vivo correlation

K i :

Inhibition constant

K p,brain :

Brain-to-plasma ratio

MDCKII-BCRP:

Madin–Darby canine kidney cells transfected with human BCRP gene

P app :

Apparent permeability coefficient

P-gp (ABCB1):

P-glycoprotein

RA:

Residual activity (% of control)

TB:

Transport buffer

TB-BSA:

Transport buffer with BSA

UPLC:

Ultra-performance liquid chromatography

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Acknowledgements and Disclosures

We thank Dr. Gerald E. Smyth, Discovery Research Laboratories in Tsukuba, Nippon Shinyaku Co., Ltd., for his help with the manuscript.

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Authors and Affiliations

  1. Pharmacokinetics and Safety Assessment Department, Discovery Research Laboratories, Nippon Shinyaku Co., Ltd, Kyoto, Japan

    Shunji Imai, Toshiki Arai, Tetsuhiro Yamada & Makoto Niwa

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  1. Shunji Imai
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  2. Toshiki Arai
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  3. Tetsuhiro Yamada
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  4. Makoto Niwa
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Correspondence to Shunji Imai.

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The authors declare no competing financial interest. This study was supported by Nippon Shinyaku Co., Ltd.

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Imai, S., Arai, T., Yamada, T. et al. Improved In Vitro-In Vivo Correlation by Using the Unbound-Fraction-Adjusted IC50 for Breast Cancer Resistance Protein Inhibition. Pharm Res 37, 230 (2020). https://doi.org/10.1007/s11095-020-02954-1

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