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Karlgren, Maria
Publications (10 of 18) Show all publications
Vallianatou, T., Strittmatter, N., Nilsson, A., Shariatgorji, M., Hamm, G., Pereira, M., . . . Andrén, P. E. (2018). A mass spectrometry imaging approach for investigating how drug-drug interactions influence drug blood-brain barrier permeability. NeuroImage, 172, 808-816
Open this publication in new window or tab >>A mass spectrometry imaging approach for investigating how drug-drug interactions influence drug blood-brain barrier permeability
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2018 (English)In: NeuroImage, ISSN 1053-8119, E-ISSN 1095-9572, Vol. 172, p. 808-816Article in journal (Refereed) Published
Abstract [en]

There is a high need to develop quantitative imaging methods capable of providing detailed brain localization information of several molecular species simultaneously. In addition, extensive information on the effect of the blood-brain barrier on the penetration, distribution and efficacy of neuroactive compounds is required. Thus, we have developed a mass spectrometry imaging method to visualize and quantify the brain distribution of drugs with varying blood-brain barrier permeability. With this approach, we were able to determine blood-brain barrier transport of different drugs and define the drug distribution in very small brain structures (e.g., choroid plexus) due to the high spatial resolution provided. Simultaneously, we investigated the effect of drug-drug interactions by inhibiting the membrane transporter multidrug resistance 1 protein. We propose that the described approach can serve as a valuable analytical tool during the development of neuroactive drugs, as it can provide physiologically relevant information often neglected by traditional imaging technologies.

Keywords
Mass spectrometry imaging, Blood-brain barrier, Drug-drug interactions, Elacridar, Loperamide, Propranolol
National Category
Pharmaceutical Sciences
Identifiers
urn:nbn:se:uu:diva-353358 (URN)10.1016/j.neuroimage.2018.01.013 (DOI)000430364100067 ()29329980 (PubMedID)
Funder
Swedish Research Council, 2013-3105]Swedish Research Council, 2014-6215]Swedish Foundation for Strategic Research , RIF14-0078]AstraZenecaEU, FP7, Seventh Framework Programme, 607517The Swedish Brain FoundationScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
Available from: 2018-06-12 Created: 2018-06-12 Last updated: 2019-09-02Bibliographically approved
Skogh, A., Lesniak, A., Sköld, C., Karlgren, M., Gaugaz, F. Z., Svensson, R., . . . Johansson, A. (2018). An imidazole based H-Phe-Phe-NH2 peptidomimetic with anti-allodynic effect in spared nerve injury mice. Bioorganic & Medicinal Chemistry Letters, 28(14), 2446-2450
Open this publication in new window or tab >>An imidazole based H-Phe-Phe-NH2 peptidomimetic with anti-allodynic effect in spared nerve injury mice
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2018 (English)In: Bioorganic & Medicinal Chemistry Letters, ISSN 0960-894X, E-ISSN 1090-2120, Vol. 28, no 14, p. 2446-2450Article in journal (Refereed) Published
Abstract [en]

The dipeptide amide H-Phe-Phe-NH2 (1) that previously was identified as a ligand for the substance P 1-7 (SP1-7) binding site exerts intriguing results in animal models of neuropathic pain after central but not after peripheral administration. The dipeptide 1 is derived from stepwise modifications of the anti-nociceptive heptapeptide SP1-7 and the tetrapeptide endomorphin-2 that is also binding to the SP1-7 site. We herein report a strong anti-allodynic effect of a new H-Phe-Phe-NH2 peptidomimetic (4) comprising an imidazole ring as a bioisosteric element, in the spare nerve injury (SNI) mice model after peripheral administration. Peptidomimetic 4 was stable in plasma, displayed a fair membrane permeability and a favorable neurotoxic profile. Moreover, the effective dose (ED50) of 4 was superior as compared to gabapentin and morphine that are used in clinic.

National Category
Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-343682 (URN)10.1016/j.bmcl.2018.06.009 (DOI)000438467200020 ()29929882 (PubMedID)
Funder
Swedish Research Council, 9459
Note

Title in dissertation reference list: An Imidazole-Based H-Phe-Phe-NH2 Peptidomimetic with Anti-Allodynic Effect in Spared Nerve Injury Mice and without Neurotoxic Liability

Available from: 2018-02-28 Created: 2018-02-28 Last updated: 2018-09-24Bibliographically approved
Karlgren, M., Simoff, I., Backlund, M., Wegler, C., Keiser, M., Handin, N., . . . Artursson, P. (2017). A CRISPR-Cas9 Generated MDCK Cell Line Expressing Human MDR1 Without Endogenous Canine MDR1 (cABCB1): An Improved Tool for Drug Efflux Studies.. Journal of Pharmaceutical Sciences, 106(9), 2909-2913
Open this publication in new window or tab >>A CRISPR-Cas9 Generated MDCK Cell Line Expressing Human MDR1 Without Endogenous Canine MDR1 (cABCB1): An Improved Tool for Drug Efflux Studies.
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2017 (English)In: Journal of Pharmaceutical Sciences, ISSN 0022-3549, E-ISSN 1520-6017, Vol. 106, no 9, p. 2909-2913Article in journal (Refereed) Published
Abstract [en]

Madin-Darby canine kidney (MDCK) II cells stably transfected with transport proteins are commonly used models for drug transport studies. However, endogenous expression of especially canine MDR1 (cMDR1) confounds the interpretation of such studies. Here we have established an MDCK cell line stably overexpressing the human MDR1 transporter (hMDR1; P-glycoprotein), and used CRISPR-Cas9 gene editing to knockout the endogenous cMDR1. Genomic screening revealed the generation of a clonal cell line homozygous for a 4-nucleotide deletion in the canine ABCB1 gene leading to a frameshift and a premature stop codon. Knockout of cMDR1 expression was verified by quantitative protein analysis and functional studies showing retained activity of the human MDR1 transporter. Application of this cell line allowed unbiased reclassification of drugs previously defined as both substrates and non-substrates in different studies using commonly used MDCK-MDR1 clones. Our new MDCK-hMDR1 cell line, together with a previously developed control cell line, both with identical deletions in the canine ABCB1 gene and lack of cMDR1 expression represent excellent in vitro tools for use in drug discovery.

Keywords
ABC transporters, MDCK cells, P-glycoprotein, drug transport, efflux pumps, genomics, membrane transporter, multidrug resistance transporters, permeability, proteomics
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-343598 (URN)10.1016/j.xphs.2017.04.018 (DOI)000417339900089 ()28450237 (PubMedID)
Funder
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Available from: 2018-02-28 Created: 2018-02-28 Last updated: 2019-07-23Bibliographically 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, p. 1-12, article id 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: 2019-07-23Bibliographically 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, p. 96-96Article 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
Karlgren, M. & Bergström, C. (2016). How Physicochemical Properties of Drugs Affect Their Metabolism and Clearance. In: Alan G. E. Wilson (Ed.), New Horizons in Predictive Drug Metabolism and Pharmacokinetics: (pp. 1-26). Cambridge: Royal Society of Chemistry
Open this publication in new window or tab >>How Physicochemical Properties of Drugs Affect Their Metabolism and Clearance
2016 (English)In: New Horizons in Predictive Drug Metabolism and Pharmacokinetics / [ed] Alan G. E. Wilson, Cambridge: Royal Society of Chemistry, 2016, p. 1-26Chapter in book (Refereed)
Abstract [en]

In this chapter the transport proteins and enzymes of importance for drug clearance are discussed. The primary organ for drug metabolism is the liver and to reach the intracellular compartment of hepatocytes, orally administered drugs must cross both the intestinal wall and the cell membrane of the liver cells. Transport proteins present in the cellular membrane may facilitate or hinder the compounds crossing these cellular barriers and hence will influence to what extent compounds will reach the enzymes. Here, the enzymes and transport proteins of importance for drug clearance are discussed. The molecular features of importance for drug interactions with transport proteins and enzymes are analyzed and the possibility to predict molecular features vulnerable to enzymatic degradation is discussed. From detailed analysis of the current literature it is concluded that for interaction, both with transport proteins and enzymes, lipophilicity plays a major role. In addition to this property, molecular properties such as hydrogen bond acceptors and donors, charge, aromaticity and molecular size can be used to distinguish between routes of clearance.

Place, publisher, year, edition, pages
Cambridge: Royal Society of Chemistry, 2016
Series
RSC Drug Discovery Series, ISSN 2041-3203 ; 49
National Category
Pharmaceutical Sciences
Research subject
Pharmacokinetics and Drug Therapy
Identifiers
urn:nbn:se:uu:diva-288002 (URN)10.1039/9781782622376-00001 (DOI)2-s2.0-84952641815 (Scopus ID)978-1-84973-828-6 (ISBN)978-1-78262-237-6 (ISBN)
Available from: 2016-04-27 Created: 2016-04-27 Last updated: 2018-01-10Bibliographically approved
Vildhede, A., Mateus, A., Khan, E. K., Lai, Y., Karlgren, M., Artursson, P. & Kjellsson, M. C. (2016). Mechanistic modeling of hepatic pitavastatin disposition: a proteomics-informed bottom-up approach. 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, 56-57
Open this publication in new window or tab >>Mechanistic modeling of hepatic pitavastatin disposition: a proteomics-informed bottom-up approach
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2016 (English)In: Drug metabolism reviews (Softcover ed.), ISSN 0360-2532, E-ISSN 1097-9883, Vol. 48, p. 56-57Article in journal, Meeting abstract (Other academic) Published
National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-303429 (URN)000380744900118 ()
Conference
20th North American Meeting of the International-Society-for-the-Study-of-Xenobiotics (ISSX), OCT 18-22, 2015, Orlando, FL
Available from: 2016-11-21 Created: 2016-09-19 Last updated: 2018-01-13Bibliographically approved
Vildhede, A., Mateus, A., Khan, E. K., Lai, Y., Karlgren, M., Artursson, P. & Kjellsson, M. C. (2016). Mechanistic Modeling of Pitavastatin Disposition in Sandwich-Cultured Human Hepatocytes: A Proteomics-Informed Bottom-Up Approach. Drug Metabolism And Disposition, 44(4), 505-516
Open this publication in new window or tab >>Mechanistic Modeling of Pitavastatin Disposition in Sandwich-Cultured Human Hepatocytes: A Proteomics-Informed Bottom-Up Approach
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2016 (English)In: Drug Metabolism And Disposition, ISSN 0090-9556, E-ISSN 1521-009X, Vol. 44, no 4, p. 505-516Article in journal (Refereed) Published
Abstract [en]

Isolated human hepatocytes are commonly used to predict transporter-mediated clearance in vivo. Such predictions, however, do not provide estimations of transporter contributions and consequently do not allow predictions of the outcome resulting from a change in the activity of a certain transporter, e.g., by inhibition or a genetic variant with reduced function. Pitavastatin is a drug that is heavily dependent on hepatic transporters for its elimination and it is mainly excreted as unchanged drug in the bile. For this reason, pitavastatin was used as a model drug to demonstrate the applicability of a bottom-up approach to predict transporter-mediated disposition in sandwich-cultured human hepatocytes (SCHH), allowing for the estimation of transporter contributions. Transport experiments in transfected HEK293 cells and inverted membrane vesicles overexpressing each of the relevant transport proteins were used to generate parameter estimates for the mechanistic model. By adjusting for differences in transporter abundance between the in vitro systems and individual SCHH batches, the model successfully predicted time profiles of medium and intracellular accumulation. Our predictions of pitavastatin bile accumulation could, however, not be confirmed due to a very low biliary excretion of pitavastatin in relation to the hepatic uptake in our SCHH. This study is, to our knowledge, the first to successfully simulate transporter-mediated processes in a complex system such as SCHH at the level of individual transport proteins using a bottom-up approach.

Keywords
pitavastatin, SCHH, hepatocytes, drug transport, OATP, NTCP, hepatic uptake, Pgp, BCRP, MRP2, MRP3
National Category
Pharmaceutical Sciences
Research subject
Biopharmaceutics
Identifiers
urn:nbn:se:uu:diva-241375 (URN)10.1124/dmd.115.066746 (DOI)000372880600005 ()26842596 (PubMedID)
Funder
Swedish Research Council, 2822Lars Hierta Memorial Foundation
Available from: 2015-01-12 Created: 2015-01-12 Last updated: 2018-01-11Bibliographically approved
Pellegrini, P., Dyczynski, M., Sbrana, F. V., Karlgren, M., Buoncervello, M., Hagg-Olofsson, M., . . . De Milito, A. (2016). Tumor acidosis enhances cytotoxic effects and autophagy inhibition by salinomycin on cancer cell lines and cancer stem cells. OncoTarget, 7(24), 35703-35723
Open this publication in new window or tab >>Tumor acidosis enhances cytotoxic effects and autophagy inhibition by salinomycin on cancer cell lines and cancer stem cells
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2016 (English)In: OncoTarget, ISSN 1949-2553, E-ISSN 1949-2553, Vol. 7, no 24, p. 35703-35723Article in journal (Refereed) Published
Abstract [en]

Sustained autophagy contributes to the metabolic adaptation of cancer cells to hypoxic and acidic microenvironments. Since cells in such environments are resistant to conventional cytotoxic drugs, inhibition of autophagy represents a promising therapeutic strategy in clinical oncology. We previously reported that the efficacy of hydroxychloroquine (HCQ), an autophagy inhibitor under clinical investigation is strongly impaired in acidic tumor environments, due to poor uptake of the drug, a phenomenon widely associated with drug resistance towards many weak bases. In this study we identified salinomycin (SAL) as a potent inhibitor of autophagy and cytotoxic agent effective on several cancer cell lines under conditions of transient and chronic acidosis. Since SAL has been reported to specifically target cancer-stem cells (CSC), we used an established model of breast CSC and CSC derived from breast cancer patients to examine whether this specificity may be associated with autophagy inhibition. We indeed found that CSC-like cells are more sensitive to autophagy inhibition compared to cells not expressing CSC markers. We also report that the ability of SAL to inhibit mammosphere formation from CSC-like cells was dramatically enhanced in acidic conditions. We propose that the development and use of clinically suitable SAL derivatives may result in improved autophagy inhibition in cancer cells and CSC in the acidic tumor microenvironment and lead to clinical benefits.

Keywords
autophagy, chloroquine, tumor acidosis, cancer therapy, pH
National Category
Cancer and Oncology
Identifiers
urn:nbn:se:uu:diva-299572 (URN)10.18632/oncotarget.9601 (DOI)000377756800018 ()
Funder
Swedish Cancer Society, CAN 2012/415
Available from: 2016-07-22 Created: 2016-07-22 Last updated: 2017-11-28Bibliographically approved
Vildhede, A., Wisniewski, J. R., Noren, A., Karlgren, M. & Artursson, P. (2015). Comparative Proteomic Analysis of Human Liver Tissue and Isolated Hepatocytes with a Focus on Proteins Determining Drug Exposure. Journal of Proteome Research, 14(8), 3305-3314
Open this publication in new window or tab >>Comparative Proteomic Analysis of Human Liver Tissue and Isolated Hepatocytes with a Focus on Proteins Determining Drug Exposure
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2015 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 14, no 8, p. 3305-3314Article in journal (Refereed) Published
Abstract [en]

Freshly isolated human hepatocytes are considered the gold standard for in vitro studies of liver functions, including drug transport, metabolism, and toxicity. For accurate predictions of the in vivo outcome, the isolated hepatocytes should reflect the phenotype of their in vivo counterpart, i.e., hepatocytes in human liver tissue. Here, we quantified and compared the membrane proteomes of freshly isolated hepatocytes and human liver tissue using a label-free shotgun proteomics approach. A total of 5144 unique proteins were identified, spanning over 6 orders of magnitude in abundance. There was a good global correlation in protein abundance. However, the expression of many plasma membrane proteins was lower in the isolated hepatocytes than in the liver tissue. This included transport proteins that determine hepatocyte exposure to many drugs and endogenous compounds. Pathway analysis of the differentially expressed proteins confirmed that hepatocytes are exposed to oxidative stress during isolation and suggested that plasma membrane proteins were degraded via the protein ubiquitination pathway. Finally, using pitavastatin as an example, we show how protein quantifications can improve in vitro predictions of in vivo liver clearance. We tentatively conclude that our data set will be a useful resource for improved hepatocyte predictions of the in vivo outcome.

Keywords
quantitative proteomics, total protein approach, oxidative stress, human hepatocytes, membrane proteome, ADME proteins, CYP enzymes, UGT enzymes, drug transporters, in vitro-in vivo extrapolation
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-261985 (URN)10.1021/acs.jproteome.5b00334 (DOI)000359394000025 ()26167961 (PubMedID)
Funder
Swedish Research Council, 2822
Available from: 2015-09-08 Created: 2015-09-07 Last updated: 2017-12-04Bibliographically approved
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