TY - JOUR A1 - Rakers, Christin A1 - Schumacher, Fabian A1 - Meinl, Walter A1 - Glatt, Hansruedi A1 - Kleuser, Burkhard A1 - Wolber, Gerhard T1 - In Silico Prediction of Human Sulfotransferase 1E1 Activity Guided by Pharmacophores from Molecular Dynamics Simulations JF - The journal of biological chemistry N2 - Acting during phase II metabolism, sulfotransferases (SULTs) serve detoxification by transforming a broad spectrum of compounds from pharmaceutical, nutritional, or environmental sources into more easily excretable metabolites. However, SULT activity has also been shown to promote formation of reactive metabolites that may have genotoxic effects. SULT subtype 1E1 (SULT1E1) was identified as a key player in estrogen homeostasis, which is involved in many physiological processes and the pathogenesis of breast and endometrial cancer. The development of an in silico prediction model for SULT1E1 ligands would therefore support the development of metabolically inert drugs and help to assess health risks related to hormonal imbalances. Here, we report on a novel approach to develop a model that enables prediction of substrates and inhibitors of SULT1E1. Molecular dynamics simulations were performed to investigate enzyme flexibility and sample protein conformations. Pharmacophores were developed that served as a cornerstone of the model, and machine learning techniques were applied for prediction refinement. The prediction model was used to screen the DrugBank (a database of experimental and approved drugs): 28% of the predicted hits were reported in literature as ligands of SULT1E1. From the remaining hits, a selection of nine molecules was subjected to biochemical assay validation and experimental results were in accordance with the in silico prediction of SULT1E1 inhibitors and substrates, thus affirming our prediction hypotheses. KW - drug design KW - drug metabolism KW - liver metabolism KW - molecular dynamics KW - molecular modeling KW - sulfotransferase Y1 - 2016 U6 - https://doi.org/10.1074/jbc.M115.685610 SN - 0021-9258 SN - 1083-351X VL - 291 SP - 58 EP - 71 PB - American Society for Biochemistry and Molecular Biology CY - Bethesda ER - TY - GEN A1 - Doege, N. A1 - Hoenzke, S. A1 - Schumacher, Fabian A1 - Balzus, Benjamin A1 - Colombo, Miriam A1 - Hadam, S. A1 - Rancan, F. A1 - Blume-Peytavi, Ulrike A1 - Schindler, A. A1 - Ruehl, E. A1 - Skov, P. A1 - Church, Martin K. A1 - Hedtrich, Sarah A1 - Kleuser, Burkhard A1 - Bodmeier, Roland A1 - Vogt, A. T1 - Ex vivo microdialysis used for the preclinical assessment of anti-inflammatory therapy T2 - Experimental dermatology : the official journal of the European Immunodermatology Society Y1 - 2016 SN - 0906-6705 SN - 1600-0625 VL - 25 SP - E32 EP - E32 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Döge, Nadine A1 - Hönzke, Stefan A1 - Schumacher, Fabian A1 - Balzus, Benjamin A1 - Colombo, Miriam A1 - Hadam, Sabrina A1 - Rancan, Fiorenza A1 - Blume-Peytavi, Ulrike A1 - Schäfer-Korting, Monika A1 - Schindler, Anke A1 - Rühl, Eckart A1 - Skov, Per Stahl A1 - Church, Martin K. A1 - Hedtrich, Sarah A1 - Kleuser, Burkhard A1 - Bodmeier, Roland A1 - Vogt, Annika T1 - Ethyl cellulose nanocarriers and nanocrystals differentially deliver dexamethasone into intact, tape-stripped or sodium lauryl sulfate-exposed ex vivo human skin - assessment by intradermal microdialysis and extraction from the different skin layers JF - Journal of controlled release N2 - Understanding penetration not only in intact, but also in lesional skin with impaired skin barrier function is important, in order to explore the surplus value of nanoparticle-based drug delivery for anti-inflammatory dermatotherapy. Herein, short-termex vivo cultures of (i) intact human skin, (ii) skin pretreated with tape-strippings and (iii) skin pre-exposed to sodium lauryl sulfate (SLS) were used to assess the penetration of dexamethasone (Dex). Intradermal microdialysis was utilized for up to 24 h after drug application as commercial cream, nanocrystals or ethyl cellulose nanocarriers applied at the therapeutic concentration of 0.05%, respectively. In addition, Dex was assessed in culture media and extracts from stratum corneum, epidermis and dermis after 24 h, and the results were compared to those in heat-separated split skin from studies in Franz diffusion cells. Providing fast drug release, nanocrystals significantly accelerated the penetration of Dex. In contrast to the application of cream and ethyl cellulose nanocarriers, Dex was already detectable in eluates after 6 h when applying nanocrystals on intact skin. Disruption of the skin barrier further accelerated and enhanced the penetration. Encapsulation in ethyl cellulose nanocarriers delayed Dex penetration. Interestingly, for all formulations highly increased concentrations in the dialysate were observed in tape-stripped skin, whereas the extent of enhancement was less in SLS-exposed skin. The results were confirmed in tissue extracts and were in line with the predictions made by in vitro release studies and ex vivo Franz diffusion cell experiments. The use of 45 kDa probes further enabled the collection of inflammatory cytokines. However, the estimation of glucocorticoid efficacy by Interleukin (IL)-6 and IL-8 analysis was limited due to the trauma induced by the probe insertion. Ex vivo intradermal microdialysis combined with culture media analysis provides an effective, skin-sparing method for preclinical assessment of novel drug delivery systems at therapeutic doses in models of diseased skin. (C) 2016 Elsevier B.V. All rights reserved. KW - Drug delivery systems KW - Polymeric nanoparticles KW - Dexamethasone KW - Microdialysis KW - Skin penetration KW - Skin barrier disruption Y1 - 2016 U6 - https://doi.org/10.1016/j.jconrel.2016.07.009 SN - 0168-3659 SN - 1873-4995 VL - 242 SP - 25 EP - 34 PB - Elsevier CY - Amsterdam ER -