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BETA
Artursson, Per
Alternative names
Publications (10 of 90) Show all publications
Beloqui, A., Brayden, D. J., Artursson, P., Preat, V. & des Rieux, A. (2017). A human intestinal M-cell-like model for investigating particle, antigen and microorganism translocation. Nature Protocols, 12(7), 1387-1399.
Open this publication in new window or tab >>A human intestinal M-cell-like model for investigating particle, antigen and microorganism translocation
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2017 (English)In: Nature Protocols, ISSN 1754-2189, E-ISSN 1750-2799, Vol. 12, no 7, 1387-1399 p.Article in journal (Refereed) Published
Abstract [en]

The specialized microfold cells (M cells) in the follicle-associated epithelium (FAE) of intestinal Peyer's patches serve as antigen-sampling cells of the intestinal innate immune system. Unlike 'classical' enterocytes, they are able to translocate diverse particulates without digesting them. They act as pathways for microorganism invasion and mediate food tolerance by transcellular transport of intestinal microbiota and antigens. Their ability to transcytose intact particles can be used to develop oral drug delivery and oral immunization strategies. This protocol describes a reproducible and versatile human M-cell-like in vitro model. This model can be exploited to evaluate M-cell transport of microparticles and nanoparticles for protein, drug or vaccine delivery and to study bacterial adherence and translocation across M cells. The inverted in vitro M-cell model consists of three main steps. First, Caco-2 cells are seeded at the apical side of the inserts. Second, the inserts are inverted and B lymphocytes are seeded at the basolateral side of the inserts. Third, the conversion to M cells is assessed. Although various M-cell culture systems exist, this model provides several advantages over the rest: (i) it is based on coculture with well-established differentiated human cell lines; (ii) it is reproducible under the conditions described herein; (iii) it can be easily mastered; and (iv) it does not require the isolation of primary cells or the use of animals. The protocol requires skills in cell culture and microscopy analysis. The model is obtained after 3 weeks, and transport experiments across the differentiated model can be carried out over periods of up to 10 h.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP, 2017
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-328973 (URN)10.1038/nprot.2017.041 (DOI)000403344900005 ()28617450 (PubMedID)
Funder
EU, FP7, Seventh Framework Programme, 281035
Available from: 2017-09-12 Created: 2017-09-12 Last updated: 2017-09-12Bibliographically approved
Llona-Minguez, S., Höglund, A., Wiita, E., Almlof, I., Mateus, A., Calderon-Montano, J. M., . . . Helledayt, T. (2017). Identification of Triazolothiadiazoles as Potent Inhibitors of the dCTP Pyrophosphatase 1. Journal of Medicinal Chemistry, 60(5), 2148-2154.
Open this publication in new window or tab >>Identification of Triazolothiadiazoles as Potent Inhibitors of the dCTP Pyrophosphatase 1
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2017 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 60, no 5, 2148-2154 p.Article in journal (Refereed) Published
Abstract [en]

The dCTP pyrophosphatase 1 (dCTPase) is involved in the regulation of the cellular dNTP pool and has been linked to cancer progression. Here we report on the discovery of a series of 3,6-disubstituted triazolothiadiazoles as potent dCTPase inhibitors. Compounds 16 and 18 display good correlation between enzymatic inhibition and target engagement, together with efficacy in a cellular synergy model, deeming them as a promising starting point for hit -to-lead development.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC, 2017
National Category
Pharmaceutical Sciences
Identifiers
urn:nbn:se:uu:diva-319535 (URN)10.1021/acsjmedchem.6b01786 (DOI)000396296100037 ()28145708 (PubMedID)
Funder
Knut and Alice Wallenberg FoundationSwedish Research CouncilEU, European Research CouncilGöran Gustafsson Foundation for promotion of scientific research at Uppala University and Royal Institute of TechnologySwedish Cancer SocietySwedish Childhood Cancer FoundationTorsten Söderbergs stiftelseRagnar Söderbergs stiftelse
Available from: 2017-04-06 Created: 2017-04-06 Last updated: 2018-01-13Bibliographically approved
Mateus, A., Treyer, A., Wegler, C., Karlgren, M., Matsson, P. & Artursson, P. (2017). Intracellular drug bioavailability: a new predictor of system dependent drug disposition. Scientific Reports, 7, 1-12, Article ID 43047.
Open this publication in new window or tab >>Intracellular drug bioavailability: a new predictor of system dependent drug disposition
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2017 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 7, 1-12 p., 43047Article in journal (Refereed) Published
Abstract [en]

Intracellular drug exposure is influenced by cell-and tissue-dependent expression of drug-transporting proteins and metabolizing enzymes. Here, we introduce the concept of intracellular bioavailability (F-ic) as the fraction of extracellular drug available to bind intracellular targets, and we assess how Fic is affected by cellular drug disposition processes. We first investigated the impact of two essential drug transporters separately, one influx transporter (OATP1B1; SLCO1B1) and one efflux transporter (P-gp; ABCB1), in cells overexpressing these proteins. We showed that OATP1B1 increased Fic of its substrates, while P-gp decreased Fic. We then investigated the impact of the concerted action of multiple transporters and metabolizing enzymes in freshly-isolated human hepatocytes in culture configurations with different levels of expression and activity of these proteins. We observed that Fic was up to 35-fold lower in the configuration with high expression of drug-eliminating transporters and enzymes. We conclude that Fic provides a measurement of the net impact of all cellular drug disposition processes on intracellular bioavailable drug levels. Importantly, no prior knowledge of the involved drug distribution pathways is required, allowing for high-throughput determination of drug access to intracellular targets in highly defined cell systems (e.g., single-transporter transfectants) or in complex ones (including primary human cells).

National Category
Medical Biotechnology
Identifiers
urn:nbn:se:uu:diva-317940 (URN)10.1038/srep43047 (DOI)000394530900001 ()28225057 (PubMedID)
Available from: 2017-04-01 Created: 2017-04-01 Last updated: 2017-11-29Bibliographically approved
Llona-Minguez, S., Höglund, A., Ghassemian, A., Desroses, M., Calderon-Montano, J. M., Moron, E. B., . . . Helleday, T. (2017). Piperazin-1-ylpyridazine Derivatives Are a Novel Class of Human dCTP Pyrophosphatase 1 Inhibitors. Journal of Medicinal Chemistry, 60(10), 4279-4292.
Open this publication in new window or tab >>Piperazin-1-ylpyridazine Derivatives Are a Novel Class of Human dCTP Pyrophosphatase 1 Inhibitors
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2017 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 60, no 10, 4279-4292 p.Article in journal (Refereed) Published
Abstract [en]

The dCTP pyrophosphatase 1 (dCTPase) is a nucleotide pool "housekeeping" enzyme responsible for the catabolism of canonical and noncanonical nucleoside triphosphates (dNTPs) and has been associated with cancer progression and cancer cell sternness. We have identified a series of piperazin-1-ylpyridazines as a new class of potent dCTPase inhibitors. Lead compounds increase dCTPase thermal and protease stability, display outstanding selectivity over related enzymes and synergize with a cytidine analogue against leukemic cells. This new class of dCTPase inhibitors lays the first stone toward the development of drug-like probes for the dCTPase enzyme.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC, 2017
National Category
Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-326233 (URN)10.1021/acs.jmedchem.7b00182 (DOI)000402498200013 ()28508636 (PubMedID)
Funder
Knut and Alice Wallenberg FoundationSwedish Research CouncilEU, European Research CouncilGöran Gustafsson Foundation for Research in Natural Sciences and MedicineSwedish Cancer SocietySwedish Childhood Cancer FoundationTorsten Söderbergs stiftelseRagnar Söderbergs stiftelse
Available from: 2017-08-10 Created: 2017-08-10 Last updated: 2018-01-13Bibliographically approved
Mateus, A., Gordon, L. J., Wayne, G. J., Almqvist, H., Axelsson, H., Seashore-Ludlow, B., . . . Artursson, P. (2017). Prediction of intracellular exposure bridges the gap between target- and cell-based drug discovery. Proceedings of the National Academy of Sciences of the United States of America, 114(30), E6231-E6239.
Open this publication in new window or tab >>Prediction of intracellular exposure bridges the gap between target- and cell-based drug discovery
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2017 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 114, no 30, E6231-E6239 p.Article in journal (Refereed) Published
Abstract [en]

Inadequate target exposure is a major cause of high attrition in drug discovery. Here, we show that a label-free method for quantifying the intracellular bioavailability (F-ic) of drug molecules predicts drug access to intracellular targets and hence, pharmacological effect. We determined F-ic in multiple cellular assays and cell types representing different targets from a number of therapeutic areas, including cancer, inflammation, and dementia. Both cytosolic targets and targets localized in subcellular compartments were investigated. F-ic gives insights on membrane-permeable compounds in terms of cellular potency and intracellular target engagement, compared with biochemical potency measurements alone. Knowledge of the amount of drug that is locally available to bind intracellular targets provides a powerful tool for compound selection in early drug discovery.

Place, publisher, year, edition, pages
NATL ACAD SCIENCES, 2017
Keyword
intracellular drug bioavailability, drug exposure, target engagement, published kinase inhibitor set, MAPK14
National Category
Pharmaceutical Sciences
Identifiers
urn:nbn:se:uu:diva-332843 (URN)10.1073/pnas.1701848114 (DOI)000406189900026 ()28701380 (PubMedID)
Funder
Swedish Research Council, 2822Carl Tryggers foundation Magnus Bergvall FoundationÅke Wiberg FoundationScience for Life Laboratory - a national resource center for high-throughput molecular bioscienceEU, FP7, Seventh Framework Programme, 607517
Available from: 2017-11-09 Created: 2017-11-09 Last updated: 2018-01-13Bibliographically approved
Llona-Minguez, S., Ghassemian, A., Baranczewski, P., Desroses, M., Koolmeister, T., Artursson, P., . . . Helleday, T. (2017). Structure-metabolism-relationships in the microsomal clearance of piperazin-1-ylpyridazines. MedChemComm, 8(7), 1553-1560.
Open this publication in new window or tab >>Structure-metabolism-relationships in the microsomal clearance of piperazin-1-ylpyridazines
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2017 (English)In: MedChemComm, ISSN 2040-2503, E-ISSN 2040-2511, Vol. 8, no 7, 1553-1560 p.Article in journal (Refereed) Published
Abstract [en]

In this study, we provide insight into the metabolic profile of a series of piperazin-1-ylpyridazines suffering from rapid in vitro intrinsic clearance in a metabolic stability assay using liver microsomes (e.g. compound 1 MLM/HLM t(1/2) = 2/3 min). Aided by empirical metabolite identification and computational predictive models, we designed the structural modifications required to improve in vitro intrinsic clearance by more than 50-fold (e.g. compound 29 MLM/HLM t(1/2) = 113/105 min).

National Category
Biochemistry and Molecular Biology Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-332436 (URN)10.1039/c7md00230k (DOI)000406090200019 ()
Funder
Swedish Research CouncilKnut and Alice Wallenberg FoundationGöran Gustafsson Foundation for Research in Natural Sciences and MedicineTorsten Söderbergs stiftelseRagnar Söderbergs stiftelseSwedish Cancer SocietySwedish Childhood Cancer Foundation
Note

De 3 första författarna delar förstaförfattarskapet.

Available from: 2017-10-31 Created: 2017-10-31 Last updated: 2018-01-13Bibliographically approved
Pedersen, J. M., Khan, E. K., Bergström, C., Palm, J., Hoogstraate, J. & Artursson, P. (2017). Substrate and method dependent inhibition of three ABC-transporters (MDR1, BCRP, and MRP2). European Journal of Pharmaceutical Sciences, 103, 70-76.
Open this publication in new window or tab >>Substrate and method dependent inhibition of three ABC-transporters (MDR1, BCRP, and MRP2)
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2017 (English)In: European Journal of Pharmaceutical Sciences, ISSN 0928-0987, E-ISSN 1879-0720, Vol. 103, 70-76 p.Article in journal (Refereed) Published
Abstract [en]

Drug transport and drug-drug interactions (DDI) with human ABC transporters are generally investigated in mammalian cell lines or inverted membrane vesicles from insect cells (Sf9) overexpressing the transporter of interest. In this study, we instead used membrane vesicles from human embryonic kidney cells (HEK293) overexpressing wild type MDR1/Pgp (ABCB1), BCRP (ABCG2), and MRP2 (ABCC2) with the aim to study the concentration dependent inhibition of shared and prototypic probe substrates. We first investigated 15 substrates and identified estrone-17-beta-glucorinide (E17G) as shared substrate. Nine specific and general inhibitors were then studied using El7G and prototypic probe substrates. The results were compared with those previously obtained in Sf9 vesicles and cell lines of canine (MDCKII) and human (Saos-2) origin. For the majority of inhibitors, K-i; values differed <10-fold between EI7G and probe substrates. Significant differences in K-i; values were observed for about one third of the inhibitors. The transport inhibition potencies in HEK293 vesicles were in good agreement with those obtained in Sf9 vesicles. Large differences were found in the inhibition potencies observed in the vesicular systems compared to the cellular systems. Nevertheless, the rank order correlations between the different experimental systems were generally good. Our study provides further information on substrate dependent inhibition of ABC-transporters, and suggests that simple ranking of compounds can be used as a tier one approach to bridge results obtained in different experimental systems.

Place, publisher, year, edition, pages
ELSEVIER SCIENCE BV, 2017
Keyword
Transport protein, ATP binding cassette transporter, Transport inhibition, DDI drug-drug-interaction, Membrane vesicles, HEK293
National Category
Pharmaceutical Sciences
Identifiers
urn:nbn:se:uu:diva-326226 (URN)10.1016/j.ejps.2017.03.002 (DOI)000402349700009 ()28263911 (PubMedID)
Funder
Swedish Research Council, 521-2009-4085, 521-2009-2651
Available from: 2017-07-04 Created: 2017-07-04 Last updated: 2018-01-13Bibliographically approved
Wegler, C., Gaugaz, F. Z., Andersson, T. B., Wisniewsk, J. R., Busch, D., Gröer, C., . . . Artursson, P. (2017). Variability in Mass Spectrometry-based Quantification of Clinically Relevant Drug Transporters and Drug Metabolizing Enzymes. Molecular Pharmaceutics, 14(9), 3142-3151.
Open this publication in new window or tab >>Variability in Mass Spectrometry-based Quantification of Clinically Relevant Drug Transporters and Drug Metabolizing Enzymes
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2017 (English)In: Molecular Pharmaceutics, ISSN 1543-8384, E-ISSN 1543-8392, Vol. 14, no 9, 3142-3151 p.Article in journal (Refereed) Published
Abstract [en]

Many different methods are used for mass-spectrometry-based protein quantification in pharmacokinetics and systems pharmacology. It has not been established to what extent the results from these various methods are comparable. Here, we compared six different mass spectrometry-based proteomics methods by measuring the expression of clinically relevant drug transporters and metabolizing enzymes in human liver. Mean protein concentrations were in general quantified to similar levels by methods using whole tissue lysates. Methods using subcellular membrane fractionation gave incomplete enrichment of the proteins. When the enriched proteins were adjusted to levels in whole tissue lysates, they were on average 4 fold lower than those quantified directly in whole tissue lysates. The differences in protein levels were propagated into differences in predictions of hepatic clearance. In conclusion, caution is needed when comparing and applying quantitative proteomics data obtained with different methods, especially since membrane fractionation is common practice for protein quantification used in drug clearance predictions.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC, 2017
Keyword
drug transporters, drug metabolizing enzymes, membrane proteins, protein quantification, targeted proteomics, label-free proteomics
National Category
Pharmaceutical Sciences
Identifiers
urn:nbn:se:uu:diva-335414 (URN)10.1021/acs.molpharmaceut.7b00364 (DOI)000410005100027 ()28767254 (PubMedID)
Funder
Swedish Research Council, 2822, 5715
Available from: 2017-12-06 Created: 2017-12-06 Last updated: 2018-01-13Bibliographically approved
Almqvist, H., Axelsson, H., Jafari, R., Dan, C., Mateus, A., Haraldsson, M., . . . Nordlund, P. (2016). CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil. Nature Communications, 7, Article ID 11040.
Open this publication in new window or tab >>CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil
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2016 (English)In: Nature Communications, ISSN 2041-1723, E-ISSN 2041-1723, Vol. 7, 11040Article in journal (Refereed) Published
Abstract [en]

Target engagement is a critical factor for therapeutic efficacy. Assessment of compound binding to native target proteins in live cells is therefore highly desirable in all stages of drug discovery. We report here the first compound library screen based on biophysical measurements of intracellular target binding, exemplified by human thymidylate synthase (TS). The screen selected accurately for all the tested known drugs acting on TS. We also identified TS inhibitors with novel chemistry and marketed drugs that were not previously known to target TS, including the DNA methyltransferase inhibitor decitabine. By following the cellular uptake and enzymatic conversion of known drugs we correlated the appearance of active metabolites over time with intracellular target engagement. These data distinguished a much slower activation of 5-fluorouracil when compared with nucleoside-based drugs. The approach establishes efficient means to associate drug uptake and activation with target binding during drug discovery.

National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-276077 (URN)10.1038/ncomms11040 (DOI)000372887500001 ()27010513 (PubMedID)
Funder
The Karolinska Institutet's Research FoundationSwedish Research CouncilSwedish Cancer SocietyKnut and Alice Wallenberg Foundation
Note

Artursson, P., Martinez-Molina, D och Nordlund, P. delar sistaförfattarskapet.

Available from: 2016-02-09 Created: 2016-02-09 Last updated: 2018-01-10Bibliographically approved
Vildhede, A., Wisniewski, J., Norén, A., Karlgren, M. & Artursson, P. (2016). Comparative proteome analysis of human liver tissue and isolated hepatocytes: application to predictions of hepatic pitavastatin uptake clearance. Paper presented at 20th North American Meeting of the International-Society-for-the-Study-of-Xenobiotics (ISSX), OCT 18-22, 2015, Orlando, FL. Drug metabolism reviews (Softcover ed.), 48, 96-96.
Open this publication in new window or tab >>Comparative proteome analysis of human liver tissue and isolated hepatocytes: application to predictions of hepatic pitavastatin uptake clearance
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2016 (English)In: Drug metabolism reviews (Softcover ed.), ISSN 0360-2532, E-ISSN 1097-9883, Vol. 48, 96-96 p.Article in journal, Meeting abstract (Other academic) Published
National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-303430 (URN)000380744900197 ()
Conference
20th North American Meeting of the International-Society-for-the-Study-of-Xenobiotics (ISSX), OCT 18-22, 2015, Orlando, FL
Note

Supplement: 1, Meeting Abstract: P143

Available from: 2016-11-21 Created: 2016-09-19 Last updated: 2018-01-13Bibliographically approved
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