For clarity only residues that have major interactions with the ligand are shown

For clarity only residues that have major interactions with the ligand are shown. inhibiting test compounds over 17-HSD1 and 17-HSD2. The selected test compounds at a concentration of 1 1 M were analyzed for their ability to inhibit 17-HSD1-dependent conversion of 200 nM estrone to estradiol (A) and the 17-HSD2-dependent conversion of 200 nM estradiol to estrone (B). Apigenin (CTRL 2) and compound 22 of Vuorinen et al. [22] (CTRL 3) served as positive controls. Data represent imply SD from three impartial experiments.(TIF) pone.0171079.s004.tif (452K) GUID:?3B0A95D1-BFEE-4EB4-BD65-FF756C8E0B8E S5 Fig: Impact of UV irradiation and determined compounds on cell viability in human skin biopsies. The experiments with human full skin biopsies were performed by Cutech Biotechnology. Skin samples were treated topically with vehicle or the respective compounds (4 L of 10 M or 100 M compound applied on top of each biopsy specimen) for 6 days either in the absence of UV treatment or upon exposure to 3.0 J/cm2 or 6.0 J/cm2 UV irradiation. Cell viability was decided after 6 days using MTT. Data symbolize imply SEM from 6 human biopsies.(TIF) pone.0171079.s005.tif (621K) GUID:?18CAC132-6C22-49AA-999F-C3968981A9BD S6 Fig: Effect of cortisone on cell viability in human skin biopsies. Human skin biopsies were treated with 0.01 M, 0.1 M or 1 M cortisone for 6 days, followed by assessment of skin viability using the MTT assay. Data symbolize imply SEM from 6 samples derived from 2 different skin biopsies. ** p<0.01 vs vehicle control.(TIF) pone.0171079.s006.tif (345K) GUID:?AD025A0B-63EF-40AE-947F-5A888398E259 S1 File: Experimental section. Materials and methods for chemistry.(DOC) pone.0171079.s007.doc (180K) GUID:?993360AB-69CC-4EE7-8167-5BC3A69C64F7 Data Availability StatementAll relevant data are within the paper and its Supporting Information files. Abstract Activity and selectivity assessment of new bi-aryl amide 11-hydroxysteroid dehydrogenase 1 (11-HSD1) inhibitors, prepared in a modular manner via Suzuki cross-coupling, are explained. Several compounds inhibiting 11-HSD1 at nanomolar concentrations were identified. Compounds 2b, 3e, 7b and 12e were shown to selectively inhibit 11-HSD1 over 11-HSD2, 17-HSD1 and 17-HSD2. These inhibitors also potently inhibited 11-HSD1 activity in intact HEK-293 cells expressing the recombinant enzyme and in intact main human keratinocytes expressing endogenous 11-HSD1. Moreover, compounds 2b, 3e and 12e were tested for their activity in human skin biopsies. They were able to prevent, at least in part, both the cortisone- and the UV-mediated decreases in collagen content. Thus, inhibition of 11-HSD1 by these compounds can be further investigated to delay or prevent UV-mediated skin damage and skin aging. Introduction GW7604 With an aging population, UV-mediated GW7604 skin damage and skin aging-related diseases represent an increasing problem and there is an increasing demand for novel therapies against skin diseases [1]. Excessive exposure to UV light results in skin damage, with erythema and DNA damage, oxidative stress, and an inflammatory response with the production of pro-inflammatory mediators such as tumor necrosis factor (TNF), interleukin 6 (IL6) and interleukin 1 (IL1), and GW7604 the activation of nuclear factor-B (NF-B) [2C4]. Glucocorticoids play an important immune modulatory role and by activating glucocorticoid receptors (GR) they suppress the expression of pro-inflammatory cytokines and activation of NF-B, thereby aiding in the resolution of the inflammatory response [5]. Human skin has the capacity to produce glucocorticoids, androgens and estrogens from synthesis of cholesterol via the steroidogenic pathway [6C10]. Besides, the local concentration of cortisol is usually controlled by 11-hydroxysteroid dehydrogenase (11-HSD) enzymes, catalyzing the interconversion of active cortisol and inactive cortisone [11]. 11-HSD1 is usually a bidirectional enzyme utilizing GW7604 cofactor NADPH and functions predominantly as an oxo-reductase transforming cortisone to cortisol [12]. It is widely expressed; in skin it has been detected in keratinocytes, dermal fibroblasts and the outer root sheath of hair follicles [13]. In contrast, 11-HSD2 utilizes cofactor NAD+, oxidizes cortisol to cortisone, and is expressed in mineralocorticoid target tissues such as kidney, colon and salivary gland but also in placenta [11], and it has also been found in keratinocytes [14, 15]. The production of glucocorticoids in skin has been shown to be strongly influenced by ultraviolet (UV) radiation. On one hand it has been shown that UVB results in an activation of a dermal regulatory system analogous to that of the hypothalamus-pituitary-adrenal (HPA) axis and activation of steroidogenic synthesis of cortisol [8, 16, 17], and on the other hand UVB and UVC (but not UVA) exposure led to an enhanced expression of 11-HSD1 but experienced no effect on 11-HSD2 (which was increased by UVA) [14]. These observations Rabbit Polyclonal to OR10H1 show that UVB exposure results in.