Paracellular and passive transcellular permeability in immortalized human corneal epithelial cell culture model

Toropainen, Elisa; Ranta, Veli‐Pekka; Vellonen, Kati‐Sisko; Palmgrén, Joni; Talvitie, Anu; Laavola, Mirka; Suhonen, Pekka; Hämäläinen, Kirsi; Auriola, Seppo; Urtti, Arto

doi:10.1016/s0928-0987(03)00173-8

Cited by 82 publications

(87 citation statements)

References 30 publications

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“…10,61 In contrast, the hydrophilic markers Na-Fl and LY displayed low permeability, Papp ¼ 6.1 6 1.1 3 10 À7 and 6.8 6 0.7 3 10 À7 , respectively, which was consistent with previous reports. 61,62 High molecular weight marker FD-4, as expected, demonstrated the lowest permeability, Papp ¼ 5.7 6 3.8 3 10…”

Section: Permeation Of Model Compoundssupporting

confidence: 61%

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

Kaluzhny

Kinuthia

Truong

et al. 2018

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. The purpose of the current work was to develop a physiologically relevant, in vitro human three-dimensional (3D) corneal epithelial tissue model for use in ophthalmic drug development.METHODS. Normal human corneal epithelial cells were cultured at the air-liquid interface to produce the 3D corneal tissue model. Corneal barrier was determined by measuring transepithelial electrical resistance (TEER). Quantitative PCR arrays were utilized to investigate expression of 84 phase I/II metabolizing enzymes and 84 drug transporter genes. Permeability was evaluated using model compounds with a wide range of hydrophobicity, molecular weight, and excipients. Finally, different formulations of latanoprost and bimatoprost were administered and drug absorption and tissue viability and integrity were investigated.RESULTS. Histologic assessment and TEER of the corneal tissue model revealed tissue structure, thickness, and barrier formation (1000 6 146 XÁcm 2 ) comparable to native human corneal epithelium. The 3D corneal tissue expressed tight junctions, mucins, and key corneal epithelial detoxification enzymes. Drug-metabolizing enzyme and transporter gene expression in 3D corneal tissue and excised human corneal epithelium were highly correlated (r 2 ¼ 0.87). Coefficients of permeation for model drugs in the tissue model and excised rabbit corneas also showed a high correlation (r 2 ¼ 0.94). As expected, latanoprost and bimatoprost free acids had much lower permeability (Papp ¼ 1.2 3 10 À6 and 1.9 3 10 À6 ) than the corresponding prodrugs (Papp ¼ 2.5 3 10 À5 and 5.6 3 10 À5 ), respectively. The presence of 0.02% benzalkonium chloride in ophthalmic formulations significantly affected tissue barrier and viability.CONCLUSIONS. The newly developed 3D corneal tissue model appears to be very useful for evaluation of corneal drug permeability and safety during ophthalmic drug development.

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Section: Permeation Of Model Compoundssupporting

confidence: 61%

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

Kaluzhny

Kinuthia

Truong

et al. 2018

Invest. Ophthalmol. Vis. Sci.

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“…The relationships identified here are unsurprising, being similar to those for permeability of epithelia and biological membranes (e.g., skin, intestine, blood-brain barrier, cornea) where lipophilicity, hydrogen bonding potential, and MW are known to be key determinants. [51,55,57,58] They are also entirely consistent with Lipinski's rules for drug-like small molecules, which emphasizes the requirement for low molecular weight (<500), low numbers of hydrogen bond donors and acceptors (<5 and <10, respectively), and a log P <5 for optimal intestinal absorption, which is a balance between aqueous solubility and membrane permeability. [54] While Eqn 1 provides a potentially useful tool for estimating tumour tissue diffusion coefficients computationally, given that all the required parameter values can be readily calculated for virtual compounds, some cautions need to be sounded.…”

Section: Discussionsupporting

confidence: 70%

Prediction of Tumour Tissue Diffusion Coefficients of Hypoxia-Activated Prodrugs from Physicochemical Parameters

et al. 2008

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The therapeutic activity of anticancer agents depends critically on their ability to penetrate through tumour tissue to reach their target cells, a requirement that is especially important for hypoxia-activated prodrugs. Here we use multicellular layers (MCL) grown in vitro from HT29 colon carcinoma cells to measure tissue diffusion coefficients (D mcl ) of 67 structurally diverse benzotriazine di-N-oxides (analogues of the hypoxia-activated prodrug tirapazamine) plus four miscellaneous compounds. An algorithm was developed to predict D mcl from physicochemical parameters (molecular weight, octanol/water partition coefficient at pH 7.4, number of hydrogen bond donors and acceptors); the fitted multivariate relationship had an explained variance (R 2 ) of 0.907 and predictive power (Q 2 ) of 0.879. Using a subset of nine compounds tested as a single cassette, the algorithm was shown to apply, with some adjustment of coefficients, to MCLs from three other tumour cell lines with differing cell packing densities (SiHa, HCT8-Ea, and HCT8-Ra). The demonstrated relationships provide tools for optimizing extravascular transport of anticancer agents during lead optimization.

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“…The buffer pH in both donor and receiver chambers was 7.4 for all of the compounds tested except for ampicillin, which was dosed in the cell medium with pH adjusted to 6.5. Samples of 100 l were collected from the receiver side every 60 min for up to 4 h. Each 100-l aliquot removed was immediately replenished with an equal Primary RCE 1) Introduced by Kawazu et al (1998); 2) introduced by Chang et al (2000) Rabbit origin; TEER low (at ϳ150 ohm ⅐ cm 2 ) in model of Kawazu et al (1998) and high (at Ͼ2000 ohm ⅐ cm 2 ) in model of Chang et al (2000); used to study transport mechanism Kamazu et al, 1998Kamazu et al, , 1999Kamazu et al, , 2006Chang et al, 2000;Dey et al, 2003;Sakanaka et al, 2006 SIRC Established by Phillips et al (1966); ATCC Immortalized cell line from rabbit cornea; TEER low at ϳ100 ohm ⅐ cm 2 ; a fibroblast phenotype; used to study permeability, transporter and esterase presence Tak et al, 2001;Dey et al, 2003;Majumdar et al, 2003;Becker et al, 2008;Goskonda et al, 1999Goskonda et al, , 2000 RCE Introduced by Araki et al (1993); ECACC Immortalized cell line from albino rabbit cornea; TEER low at ϳ150 ohm ⅐ cm 2 ; used to study drug permeation Burgalassi et al, 2004 HCE-T Established by Araki-Sasaki et al (1995) Immortalized cell line by infection of human corneal epithelial cells with a recombinant simian virus 40-adenovirus vector; tight multilayered with TEER Ͼ400 ohm ⅐ cm 2 ; used to study drug transport and transporter presence Toropainen et al, 2001Toropainen et al, , 2003Ranta et al, 2003;Becker et al, 2007Becker et al, , 2008 Permeability Data and Statistical Analysis. Apparent permeability coefficients (P app ) were calculated using the following equation:…”

Section: Methodsmentioning

confidence: 99%

Characterization of Human Corneal Epithelial Cell Model As a Surrogate for Corneal Permeability Assessment: Metabolism and Transport

Xiang

Batugo²,

Gale³

et al. 2009

Drug Metab Dispos

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ABSTRACT:The recently introduced Clonetics human corneal epithelium (cHCE) cell line is considered a promising in vitro permeability model, replacing excised animal cornea to predict corneal permeability of topically administered compounds. The purpose of this study was to further characterize cHCE as a corneal permeability model from both drug metabolism and transport aspects. First, good correlation was found in the permeability values (P app ) obtained from cHCE and rabbit corneas for various ophthalmic drugs and permeability markers. Second, a previously established realtime quantitative polymerase chain reaction method was used to profile mRNA expression of drug-metabolizing enzymes (major cytochromes P450 and UDP glucuronosyltransferase 1A1) and transporters in cHCE in comparison with human cornea. Findings indicated that 1) the mRNA expression of most metabolizing enzymes tested was lower in cHCE than in excised human cornea, 2) the mRNA expression of efflux transporters [multidrug resistantassociated protein (MRP) 1, MRP2, MRP3, and breast cancer resistance protein], peptide transporters (PEPT1 and PEPT2), and organic cation transporters (OCTN1, OCTN2, OCT1, and OCT3) could be detected in cHCE as in human cornea. However, multidrug resistance (MDR) 1 and organic anion transporting polypeptide 2B1 was not detected in cHCE; 3) cHCE was demonstrated to possess both esterase and ketone reductase activities known to be present in human cornea; and 4) transport studies using probe substrates suggested that both active efflux and uptake transport may be limited in cHCE. As the first detailed report to delineate drug metabolism and transport characteristics of cHCE, this work shed light on the usefulness and potential limitations of cHCE in predicting the corneal permeability of ophthalmic drugs, including ester prodrugs, and transporter substrates.Topical instillation is the desired route of administration for ophthalmic drugs to treat diseases in the anterior segment of the eye including glaucoma, inflammations, infections, and dry eye. The primary pathway of drug permeation from the tear fluid to the anterior chamber of the eye is via the transcorneal route. The cornea has a multilayered structure constituted primarily of corneal epithelium, stroma, and endothelium. However, passage through the corneal epithelium is considered to be the rate-limiting step in the transcorneal penetration of most ophthalmic drugs (Maurice and Mishima, 1984).In the past decade, several in vitro corneal permeability models that can substitute for the isolated cornea to predict ocular absorption and facilitate ophthalmic drug discovery have been developed (Reichl and Becker, 2008). These models evolved from primary cultures using rabbit corneas and immortalized rabbit cell lines to the immortalized human corneal epithelial (HCE) cell lines, and even include complex systems such as human corneal constructs (summarized in Table 1). Given the pros and cons of each model, HCE cell lines seem to be most appealing owing to their human o...

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Paracellular and passive transcellular permeability in immortalized human corneal epithelial cell culture model

Cited by 82 publications

References 30 publications

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

Prediction of Tumour Tissue Diffusion Coefficients of Hypoxia-Activated Prodrugs from Physicochemical Parameters

Characterization of Human Corneal Epithelial Cell Model As a Surrogate for Corneal Permeability Assessment: Metabolism and Transport

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