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Human Primary Cell-Based Organotypic Microtissues for Modeling Small Intestinal Drug Absorption

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Abstract

Purpose

The study evaluates the use of new in vitro primary human cell-based organotypic small intestinal (SMI) microtissues for predicting intestinal drug absorption and drug-drug interaction.

Methods

The SMI microtissues were reconstructed using human intestinal fibroblasts and enterocytes cultured on a permeable support. To evaluate the suitability of the intestinal microtissues to model drug absorption, the permeability coefficients across the microtissues were determined for a panel of 11 benchmark drugs with known human absorption and Caco-2 permeability data. Drug-drug interactions were examined using efflux transporter substrates and inhibitors.

Results

The 3D–intestinal microtissues recapitulate the structural features and physiological barrier properties of the human small intestine. The microtissues also expressed drug transporters and metabolizing enzymes found on the intestinal wall. Functionally, the SMI microtissues were able to discriminate between low and high permeability drugs and correlated better with human absorption data (r2 = 0.91) compared to Caco-2 cells (r2 = 0.71). Finally, the functionality of efflux transporters was confirmed using efflux substrates and inhibitors which resulted in efflux ratios of >2.0 fold and by a decrease in efflux ratios following the addition of inhibitors.

Conclusion

The SMI microtissues appear to be a useful pre-clinical tool for predicting drug bioavailability of orally administered drugs.

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Abbreviations

Ω:

Ohm

2D:

Two dimensional

3D:

Three dimensional

Ǻ:

Angstrom

(A):

Apical

ABCB1:

ATP binding cassette subfamily B member 1

ABCC1:

ATP Binding Cassette Subfamily C Member 1

ABCC2:

ATP Binding Cassette Subfamily C Member 2

ABCG2:

ATP-binding cassette sub-family G member 2

ADR:

Adverse drug reaction

ALI:

Air–liquid interface

(B):

Basolateral

BCRP:

Breast cancer resistance protein

BCS:

Biopharmaceutical Classification System

CDCF:

5(6)-carboxy-2′,7′-dichlorofluorescein (CDCF)

cDNA:

Complementary DNA

CK:

Cytokeratin

Cq:

PCR cycles

Ct:

Threshold cycle

CYP450:

Cytochrome P450

DDI:

Drug-drug interaction

FDA:

Food and Drug Administration

FT:

Full-thickness

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GI:

Gastrointestinal

HPLC:

High performance liquid chromatography

H:

Hour

IIAM:

International Institute for the Advancement of Medicine

LC/MS:

Liquid chromatography–mass spectrometry

LY:

Lucifer Yellow

MDCK:

Madin-Darby canine kidney

MDR1:

Multi-drug resistance gene (MDR)-1

Met:

Metabolite

Min:

Minutes

MRP-1:

Multidrug-resistance associated protein-1

MRP-2:

Multidrug-resistance associated protein-2

MS MRM:

Mass spectroscopy multiple reaction monitoring

MTT:

3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide

OPO:

Organ Procurement Organization

Papp :

Apparent permeability coefficient

P-gp:

p-Glycoprotein

PR:

Parental

PT:

Partial thickness

qPCR:

Quantitative polymerase chain reaction

RFU:

Relative fluorescence unit

RNA:

Ribonucleic acid

RT:

Room temperature

RT-PCR:

Reverse Transcription Polymerase Chain Reaction

SEM:

Scanning electron microscopy

SMI:

Small intestine

TEER:

Transepithelial electrical resistance

TEM:

Transmission electron microscopy

TTT:

TEER of treated tissues

TUT:

TEER of untreated tissues

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Authors and Affiliations

  1. MatTek Corporation, 200 Homer Avenue, Ashland, Massachusetts, USA

    Seyoum Ayehunie, Tim Landry, Zachary Stevens, Alex Armento, Patrick Hayden & Mitchell Klausner

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  1. Seyoum Ayehunie
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Correspondence to Seyoum Ayehunie.

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Ayehunie, S., Landry, T., Stevens, Z. et al. Human Primary Cell-Based Organotypic Microtissues for Modeling Small Intestinal Drug Absorption. Pharm Res 35, 72 (2018). https://doi.org/10.1007/s11095-018-2362-0

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