The new low calcemic vitamin D analog 22-ene-25-oxa-vitamin D prominently ameliorates T helper cell type 1-mediated colitis in mice

1st Department of Internal Medicine, Division of Gastroenterology and Clinical Nutrition, Johann Wolfgang Goethe University, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany.
Journal of Pharmacology and Experimental Therapeutics (Impact Factor: 3.86). 12/2006; 319(2):622-31. DOI: 10.1124/jpet.106.107599
Source: PubMed

ABSTRACT In addition to its well defined role as a key regulator of calcium and bone metabolism, 1,25-dihydroxyvitamin D(3) (calcitriol) has been established as a potent modulator of immune cell function. Still, because of the hypercalcemic toxicity occurring after systemic application of the parent compound, its clinical application as an immunosuppressant has been hampered. Recently, we described 22-ene-25-oxa-vitamin D (ZK156979) as a representative of a novel class of low calcemic vitamin D analogs with well preserved immunosuppressive activity in vitro. Here, in vivo colitis was induced by applying a rectal enema of 2,4,6-trinitrobenzene sulfonic acid (TNBS) to male BALB/c mice, and calcitriol (0.2 microg/kg) or ZK156979 (0.1-2.0 microg/kg) was given i.p. from days 0 to 3 or 3 to 5. Body mass and clinical activity score of colitis were recorded daily. Colon tissue was analyzed macroscopically and microscopically, myeloperoxidase activity and cytokine levels [tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-10, and IL-4] were determined by enzyme-linked immunosorbent assay, and T-box transcription factor (T-bet) expression was determined by immunoblot analysis. We found that treatment with ZK156979 clearly reduced the severity of TNBS-induced colitis without exhibiting calcemic effects. Both early and late treatment abrogated body weight loss, diarrhea, and macroscopic intestinal inflammation with a potency comparable with that of calcitriol. The therapeutic effect of ZK156979 was accompanied by a down-regulation of myeloperoxidase activity, TNF-alpha, IFN-gamma, and T-bet expression decreased, whereas local tissue IL-10 and IL-4 protein levels increased. To conclude, our data provide the first clear evidence that ZK156979 exhibits a beneficial prophylactic as well as therapeutic profile in T helper cell type 1-like experimental colitis, offering new therapeutic options for the treatment of human inflammatory bowel diseases.

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Available from: Heinfried H Radeke, Nov 11, 2014
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    • "By performing modifications in the side chain of 1,25-(OH) 2 D 3 , one group described the VDR agonist ZK156979 (22-ene-25-oxa- vitamin D), which at normocalcemic doses effectively improved the symptoms of colitis induced by 2,4,6- trinitrobenzene sulfonic acid by inhibiting TNFa production and increasing levels of anti-inflammatory cytokines. In vivo experiments performed in rats showed that ZK156979 exhibited 100-fold lower hypercalcemic activity than 1,25-(OH) 2 D 3 (Daniel et al., 2005, 2006). Use of another VDR analog, TX527 [19-nor-14,20- bisepi-23-yne-1,25(OH) 2 D 3 ], proved efficacious at inhibiting cell proliferation and TNFa production in vitro in human PBMCs from Crohn's disease patients (Stio et al., 2007). "
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    • "Protein extracts from colon samples (obtained 2 and 4 cm above the anus and pooled), PBMCs, MM6 cells, and activated mesenteric lymph nodes (MLNs) were prepared by using a cell extraction kit (Active Motif Nuclear Extract Kit; Active Motif, Rixensart, Belgium) according to the instructions of the manufacturer and analyzed for protein content with bicinchoninic acid protein assay reagent (Thermo Fisher Scientific, Bonn, Germany). For SDS-polyacrylamide gel electrophoresis and Western blotting refer to the protocol described previously (Daniel et al., 2006). Visualization and quantification of protein bands were carried out with the Odyssey Infrared Imaging System (LI-COR Biosciences, Bad Homburg, Germany). "
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    • "Extending two previous investigations (Daniel et al., 2005, 2006), the data of this in vivo study in an accepted model of Th1-mediated colitis in mice clearly indicates and unravels that combining dexamethasone with calcitriol represents an attractive new immune modulatory treatment regimen of inflammatory disorders, including CD. To our knowledge, here we show for the first time that the complex immunemodulating effects of calcitriol may result from a differential down-regulation of proinflammatory capacities of intestinal DC as assessed by analysis of IL-12p70 as well as IL-23p19. "
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