cholecalciferol and calcipotriene

Within the past decade it has been shown that psoriasis can be treated topically with analogs of vitamin-D3. Impaired differentiation and increased proliferation of keratinocytes are key features in psoriatic lesions together with a local activation of T lymphocytes. Evidence has accumulated showing that analogs of vitamin D3 increase differentiation and inhibit proliferation of keratinocytes. Therefore, analogs of vitamin D3 have been investigated in a number of trials showing improvement of psoriasis. It has been shown that vitamin D analogs are better than their vehicle and show the same potency as potent topical steroids. However, vitamin D analogs have been proven efficacious and without side effects also when used on long term basis. Vitamin D analogs can be used both as monotherapy and in combination topical steroids, UVB, PUVA, retinoids and cysclosporine. The vitamin D3 analog calcipotriol has been investigated in most detail and is available as an ointment, a creme and as a scalp solutation. From clinical studies involving thousands of patients, it can be concluded that calcipotriol is efficacious, safe, well tolerated and can be used on a long term basis. Other analogs are available, however, these analogs have not been studied in greater details yet.

Topics: Calcitriol; Cholecalciferol; Dermatologic Agents; Dihydroxycholecalciferols; Drug Therapy, Combination; Humans; Psoriasis

The natural form of vitamin D3, 1,25-dihydroxyvitamin D3, modulates epidermal proliferation and differentiation and inhibits immune induction. Calcipotriol (calcipotriene) is a synthetic vitamin D analogue that partially separates the newly identified effects from the classic hypercalcemic effect. Topical treatment with calcipotriene has been shown to be effective, well tolerated, and safe for psoriasis.

Topics: Administration, Cutaneous; Calcitriol; Cell Differentiation; Cell Division; Cholecalciferol; Dermatologic Agents; Epidermis; Humans; Psoriasis

The introduction of this article is devoted to the physiological role of vitamin D3 and its synthesis. The authors refer to the accidental introduction of vitamin D3 into the psoriasis treatment, which later led to the development of a less toxic vitamin D3 analog named calcipotriol, and its administration as a topical antipsoriatic agent. Reviewing the literature the authors discuss the pharmacological and immunological effects of calcipotriol as well as the potential applications of vitamin D3.

Topics: Administration, Topical; Calcitriol; Cholecalciferol; Dermatologic Agents; Humans; Psoriasis

Topics: 25-Hydroxyvitamin D3 1-alpha-Hydroxylase; Calcitriol; Cholecalciferol; Dermatologic Agents; Humans; Hydroxycholecalciferols; Psoriasis

The naturally occurring 1,25(OH)2 vitamin D3 and the two synthetic derivatives 1,24(OH)2 vitamin D3 and calcipotriol (Calcipotriol, INN, Calcipotriene, USAN) have a profound effect on keratinocyte proliferation and differentiation. Clinical trials have shown an effect of all three derivatives in psoriasis. The limiting factor for the use of 1,25(OH)2 vitamin D3 is the risk of hypercalcemia, which is 100 to 200 times less for calcipotriol and that may be less also for 1,24(OH)2 vitamin D3. Presently, calcipotriol in an ointment base has been marketed in a number of countries and represents an alternative to treatment with topical glucocorticoids and dithranol.

Topics: Calcitriol; Cholecalciferol; Dermatologic Agents; Humans; Psoriasis

The physiologically active metabolite of vitamin D3, 1 alpha,25-dihydroxycolicalciferol [1,25(OH)2D3, calcitriol] has achieved the status of a hormone. It is believed to mediate its effects by binding to a specific receptor which belongs to the family of nuclear receptors for glucocorticoids, estrogens, thyroxine, and retinoid acid. It has been discovered that 1,25(OH)2D3 has the ability to regulate growth and differentiation in many cell types, including cancer cells, epidermal keratinocytes, and activated lymphocytes. This has set the stage for the development of a new class of compounds with potential usefulness in hyperproliferative and immune-mediated diseases. Ideally, such agents should possess potent effects as regulators of cell proliferation and differentiation at concentrations well below those that may induce side effects related to the classical vitamin D activity on calcium absorption and bone mineralization. In addition to 1,25(OH)2D3, the synthetic vitamin D3 analogues 1 alpha-OH-D3, 1,24(OH)2D3, and calcipotriol have undergone clinical evaluation. Calcipotriol has been studied most extensively. Compared with 1,25(OH)2D3, calcipotriol is about 200 times less potent in its effect on calcium metabolism although similar in receptor affinity. In double-blind, placebo-controlled multicenter studies, topical calcipotriol has been shown to be both efficacious and safe for the short- and long-term treatment of plaque-type psoriasis. Because some of the novel vitamin D analogues are potent regulators of cell growth and immune responses, they may be of potential interest in the treatment of ichthyoses, cancer, and autoimmune diseases.

Topics: Calcitriol; Cholecalciferol; Humans; Psoriasis; Skin; Vitamin D

Alopecia areata is a chronic relapsing autoimmune inflammatory hair disorder with no novel therapy. The objectives of this study are to compare the efficacy of topical calcipotriol vs narrow band ultraviolet B phototherapy (NB-UVB) in the treatment of alopecia areata and its correlation with serum vitamin D

Topics: Administration, Topical; Adolescent; Adult; Alopecia Areata; Calcitriol; Cholecalciferol; Combined Modality Therapy; Dermatologic Agents; Female; Humans; Male; Severity of Illness Index; Treatment Outcome; Ultraviolet Therapy; Young Adult

Topics: Adult; Aged; Aged, 80 and over; Calcitriol; Cholecalciferol; Dermatologic Agents; Dihydroxycholecalciferols; Female; Humans; Male; Middle Aged; Psoriasis; Treatment Outcome

Combining phototherapy with topical vitamin D3 analogues is a useful therapy for the treatment of psoriasis by reducing the cumulative UV dose required for clearance of lesions. Experimental investigations demonstrated that calcipotriol is degraded by UV radiation, and suggested that calcipotriol should be applied after phototherapy but not immediately before.. Calcipotriol or maxacalcitol ointment was topically applied to psoriatic plaques of six patients immediately before or after phototherapy on the right or left side of the body, respectively.. Topical application of vitamin D3 analogues either before or after irradiation by psoralen and UVA radiation (PUVA) or narrow-band (NB)-UVB showed exactly similar effects in all patients.. Therapeutic effects of vitamin D3 analogues are not clinically inactivated by subsequent irradiation with PUVA or NB-UVB phototherapy.

Topics: Administration, Cutaneous; Adult; Calcitriol; Cholecalciferol; Combined Modality Therapy; Dermatologic Agents; Female; Humans; Male; Middle Aged; Prospective Studies; Psoriasis; Severity of Illness Index; Treatment Outcome; Ultraviolet Therapy

Vitamin D receptor (VDR) activities have been noted for a number of B cell malignancies which showed varying sensitivities to vitamin D3 (1,25-dihydroxyvitamin D3, VD3, calcitriol) and its synthetic analogs. The objective of this study was to address the potential effects of VD3 and vitamin D3 analogs (VDAs) on the growth of Hodgkin's lymphoma (HL), a malignant pathology of B cell origin, in vitro.. Immunofluorescence staining showed the expression of VDR by primary Hodgkin's (H) and Reed-Sternberg (RS)-HRS-tumor cells in HL histological sections. Western blot analyses revealed expression of VDR in the HL cell lines Hs445, HDLM2, KMH2, and L428. One-way analysis of variance (ANOVA) on data obtained from water-soluble tetrazolium 1 (WST-1) cell proliferation assay showed decreased cell growth in HDLM2 and L428, 72 h after treatment with 10 µM of either VD3 of VDAs. Western blot analyses showed that treatment of L428 cells with the VDAs (calcipotriol and EB1089) resulted in modest increases in nuclear accumulation of VDR (nuVDR) compared to either dimethyl sulfoxide (DMSO) or VD3 treatments. nuVDR for DMSO control and VD3 was comparable. These results suggest that VD3 or VDAs may affect growth of HL.

Topics: Calcitriol; Cell Line, Tumor; Cell Proliferation; Cholecalciferol; Dimethyl Sulfoxide; Gene Expression Regulation, Neoplastic; Hodgkin Disease; Humans; Receptors, Calcitriol; Vitamin D

To study the molecular effects of three different D-vitamins, vitamin D2, vitamin D3 and calcipotriol, in pancreatic stellate cells (PSCs).. Quiescent PSCs were isolated from mouse pancreas and activated. Sustained culture of originally quiescent PSCs induced cell proliferation, loss of lipid droplets and exhibition of stress fibers, indicating cell activation. When added to PSCs in primary culture, all three D-vitamins diminished expression of α-SMA (to 32%-39% of the level of control cells;. The three D-vitamins inhibit, with similar efficiencies, activation of PSCs

Topics: Actins; Animals; Calcitriol; Cell Proliferation; Cells, Cultured; Cellular Senescence; Cholecalciferol; DNA Replication; Ergocalciferols; Fibrosis; Interleukin-6; Lipid Droplets; Mice, Inbred C57BL; Myofibroblasts; Pancreas; Pancreatic Diseases; Pancreatic Stellate Cells; Phenotype; Proline

Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disease with pruritus and high prevalence. Indeed, 15-30 % of children and 2-10 % of adults from industrialized countries are affected. Acute AD lesions are characterized by epidermal hyperplasia associated with a dominant Th2/Th17 immune response and dermal inflammatory infiltrates. Moreover, the expression of alarmins such as TSLP, IL-33, and IL-25 is upregulated in acute AD lesions. Topical application of vitamin D3 or of its low-calcemic analog MC903 induces changes in skin morphology and inflammation resembling immune perturbations observed in acute lesions of patients with AD. Mice treated with MC903 or vitamin D3 additionally display increased serum IgE levels, as observed in patients with extrinsic AD. Interestingly, these symptoms are not dependent on mouse gender or on genetic background. Thus, the easiness of this mouse model renders it very attractive to study immunologic abnormalities involved in AD development or maintenance. Furthermore, this model might be useful for preclinical studies aiming at unraveling new therapeutic strategies to treat AD. In this chapter, we describe the induction and major features of MC903 and vitamin D3-induced AD-like inflammation in mice.

Topics: Administration, Cutaneous; Animals; Biomarkers; Calcitriol; Cholecalciferol; Dermatitis, Atopic; Disease Models, Animal; Drug Administration Schedule; Ear; Female; Gene Expression; Humans; Immunoglobulins; Keratinocytes; Male; Mice; Mice, Inbred C57BL; Receptors, Cytokine; Skin

In relation to Th17 cell actions, interferon (IFN)-α production by plasmacytoid dendritic cells (pDCs) are involved in the pathogenesis of psoriasis. Vitamin D3 analogues are widely used in the treatment of psoriasis, however, their actions on pDCs are not well understood.. To investigate the effects of Vitamin D3 analogue calcipotriol (CAL) on pDCs, focusing on the cytokine production and chemotactic activity.. We compared in mice the effects of CAL, cyclosporine A (CyA), and triamcinolone acetonide (TA) on the cytokine production by pDCs (IFN-α), conventional DCs (TNF-α), and γd T cells (IL-17A). pDCs isolated from mouse spleen cells were stimulated with CpG-ODN in the presence or absence of each drug for 48h. Purified splenic conventional DCs (cDCs) and lymph node γδ T cells were stimulated with CpG-ODN or with anti-CD3/CD28 antibody, respectively. IFN-α, TNF-α and IL-17A in the 48-h culture supernatants were quantified by ELISA. We also studied the ability of CAL to inhibit the chemotaxis of freshly isolated pDCs toward chemerin and VEGF-A, representative chemoattractants of pDCs, by a real-time monitoring method, EZ-Taxiscan. To assess the effect of CAL on pDC accumulation in vivo, we painted CAL ointment to the mouse skin inflamed by topical application of imiquimod cream (IMQ) for 4 consecutive days. In the skin samples, we enumerated 440c. CAL significantly inhibited CpG-enhanced pDC IFN-α production at a comparable level to T cell IL-17A production, whereas its effect on cDC TNF-α production was minimal. Accordingly, CAL suppressed the CpG-augmented expression of TLR9 and MyD88. On the contrary, CyA strongly suppressed the production of TNF-α and IL-17A, but not IFN-α. TA inhibited the production of all the cytokines tested. The effect of CAL on the chemotactic activity of pDCs was also evaluated, demonstrating a significant downmodulation by exposure to the reagent. CAL depressed chemerin receptor CMKLR1 expression in pDCs. The in vivo mouse study showed that simultaneous application of CAL to the imiquimod-applied skin reduce both the recruitment of pDCs and the expression of IFN-α2 in the skin.. Our findings suggest that CAL uniquely downmodulates the cytokine production and chemotactic activity of pDCs. The CAL suppression of the in vivo pDC accumulation to the skin suggests that these actions are therapeutically relevant.

Topics: Animals; Calcitriol; Chemotaxis; Cholecalciferol; CpG Islands; Dendritic Cells; Dermatologic Agents; Down-Regulation; Female; Flow Cytometry; Humans; Immunohistochemistry; Interferon-alpha; Interferon-gamma; Interleukin-17; Mice; Microscopy, Fluorescence; Oligonucleotides; Psoriasis; Receptors, Antigen, T-Cell, gamma-delta; Skin; Spleen; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Current data from daily practice show that vitamin D₃ analogues, corticosteroids and their fixed combination products are used heterogeneously for topical long-term treatment of psoriasis.. To evaluate scientific evidence about topical long-term therapy with vitamin D3 analogues, corticosteroids and their two-compound-products in psoriasis vulgaris and scalp psoriasis and to develop daily practice recommendations.. Systematic literature review via PubMed and Embase and informal expert consensus.. The search strategy identified 21 relevant clinical trials. Best evidence regarding topical long term treatment was available for the two-compound-formulation containing calcipotriene and betamethasone. In a comparative trial in psoriasis vulgaris the two-compound-formulation showed superior tolerability and cost effectiveness compared to monotherapy. In scalp psoriasis the two-compound-gel was superior compared to calcipotriene monotherapy. Standardized and simplified treatment application modes resulted in a better clinical outcome comparing to on-demand therapies.. Patients should be proactively involved in the choice of treatment, formulation and mode of application. Besides long-term treatment with the two-compound-formulation other treatment regimens including calcipotriene monotherapy can also be considered. Due to a favorable risk-benefit ratio in maintenance trials and due to better cost-effectiveness the application of two-compound-products once or twice a week after initial therapy is recommended.

Topics: Administration, Topical; Adrenal Cortex Hormones; Betamethasone; Calcitriol; Cholecalciferol; Clinical Trials as Topic; Drug Combinations; Evidence-Based Medicine; Guideline Adherence; Humans; Long-Term Care; Psoriasis

We describe here the effects of three drugs that are either approved or have the potential for treating multiple sclerosis (MS) patients through the in vitro activities of human natural killer (NK) cells and dendritic cells (DCs). Our results indicate that 1,25(OH)2D3, the biologically active metabolite of vitamin D3, calcipotriol and FTY720 augment IL-2-activated NK cell lysis of K562 and RAJI tumor cell lines as well as immature (i) and mature (m) DCs, with variable efficacies. These results are corroborated with the ability of the drugs to up-regulate the expression of NK cytotoxicity receptors NKp30 and NKp44, as well as NKG2D on the surfaces of NK cells. Also, they down-regulate the expression of the killer inhibitory receptor CD158. The three drugs down-regulate the expression of CCR6 on the surface of iDCs, whereas vitamin D3 and calcipotriol tend to up-regulate the expression of CCR7 on mDCs, suggesting that they may influence the migration of DCs into the lymph nodes. Finally, vitamin D3, calcipotriol and FTY720 enhance NK17/NK1 cell lysis of K562 cells, suggesting that a possible mechanism of action for these drugs is via activating these newly described cells. In conclusion, our results show novel mechanisms of action for vitamin D3, calcipotriol and FTY720 on cells of the innate immune system.

Topics: Calcitriol; Cell Line, Tumor; Cholecalciferol; Cytotoxicity, Immunologic; Dendritic Cells; Down-Regulation; Fingolimod Hydrochloride; Gene Expression Regulation, Neoplastic; Humans; Interleukin-2; K562 Cells; Killer Cells, Natural; Membrane Proteins; Multiple Sclerosis; Natural Cytotoxicity Triggering Receptor 2; Natural Cytotoxicity Triggering Receptor 3; NK Cell Lectin-Like Receptor Subfamily K; Propylene Glycols; Receptors, CCR6; Receptors, CCR7; Sphingosine; Up-Regulation

Psoriasis is a chronic inflammatory skin disease of unknown aetiology, and an active form of vitamin D(3) (1α,25-dihydroxyvitamin D(3)) and its analogues (VD3As) are widely used topical reagents for psoriasis treatment. Besides their well-known calcium homeostasis functions, VD3As have been shown to have various immune-modulating effects including the induction of thymic stromal lymphopoietin (TSLP), a master cytokine for inducing Th2 inflammation, in mouse models, but not yet in human psoriasis. VD3As also have been shown to induce cathelicidin, an antimicrobial peptide and strong inducer of innate immunity. Cathelicidin is overexpressed in psoriatic skin lesions; however, its role in this disease seems as yet inconclusive.. To clarify whether topical VD3As induce TSLP and cathelicidin, and to examine the modulation of expression patterns of related cytokines in human psoriatic lesions.. Skin biopsy samples from psoriatic lesions with or without VD3A treatment were subjected to immunohistochemical staining and quantitative reverse transcription-polymerase chain reaction analyses to measure the expression levels of various cytokines.. Significantly higher levels of TSLP, thymus and activation-related chemokine and CCR4 expression were observed in VD3A+ skin samples than in VD3A- samples. In contrast, significantly lower levels of interleukin (IL)-12/23 p40, IL-1α, IL-1β and tumour necrosis factor (TNF)-α expression were observed in the VD3A+ samples than in the VD3A- samples. Expression of cathelicidin was elevated in VD3A+ samples.. Topical VD3As induce TSLP and cathelicidin in psoriatic lesions, resulting in suppression of IL-12/23 p40, IL-1α, IL-1β and TNF-α, thereby ameliorating psoriatic plaques.

Topics: Administration, Cutaneous; Adult; Aged; Antimicrobial Cationic Peptides; Blotting, Western; Calcitriol; Cathelicidins; Cholecalciferol; Cytokines; Dermatologic Agents; Dihydroxycholecalciferols; Drug Therapy, Combination; Female; Humans; Interleukins; Male; Middle Aged; Psoriasis; Thymic Stromal Lymphopoietin; Tumor Necrosis Factor-alpha

Peptide conjugation to the surface of stealth liposomes has been studied for liposomal drug targeting to cells expressing specific receptors to provide a site-specific drug delivery. This study investigated the potential of peptide-conjugated liposomes for targeting cells expressing the human integrin α(2)β(1) receptor. A 12 amino acid head-to-tail cyclic peptide derived from the Jararhagin protein containing the Arg-Lys-Lys-His (RKKH)-specific binding site was conjugated to the distal ends of poly(ethylene glycol) (PEG) chains on PEGylated liposomes. Epithelial cells expressing the receptor showed increased cellular association and uptake of peptide-conjugated liposomes at 4 °C, compared to liposomes conjugated with a non-specific peptide. The interaction between cells and peptide-conjugated liposomes was significantly increased at 37 °C suggesting that a possible uptake mechanism might be energy-dependent endocytosis. In keratinocyte cell cultures, the ligand-conjugated liposomes loaded with the vitamin D(3) analogue calcipotriol induced transcription of the gene encoding the antimicrobial peptide cathelicidin, which is activated through the vitamin D(3) receptor upon binding of vitamin D(3) analogues. This suggests that the liposomes are internalized and that calcipotriol is delivered intracellularly and released in an active form. In conclusion, the 12 amino acid head-to-tail cyclic RKKH peptide seems promising for targeting of liposomes to the integrin α(2)β(1) receptor.

Topics: Antimicrobial Cationic Peptides; Binding Sites; Calcitriol; Cathelicidins; Cell Line, Tumor; Cells, Cultured; Chemistry, Pharmaceutical; Cholecalciferol; Drug Delivery Systems; Endocytosis; Epithelial Cells; Humans; Integrin alpha2beta1; Keratinocytes; Ligands; Liposomes; Peptides, Cyclic; Polyethylene Glycols; Receptors, Calcitriol

Analogues of vitamin D3 are extensively used in the treatment of various illnesses, such as osteoporosis, inflammatory skin diseases, and cancer. Functional testing of new vitamin D3 analogues and formulations for improved systemic and topical administration is supported by sensitive screening methods that allow a comparative evaluation of drug properties. As a new tool in functional screening of vitamin D3 analogues, we describe a genomically integratable sensor for sensitive drug detection. This system facilitates assessment of the pharmacokinetic and pharmadynamic properties of vitamin D3 analogues. The tri-cistronic genetic sensor encodes a drug-sensoring protein, a reporter protein expressed from an activated sensor-responsive promoter, and a resistance marker.. The three expression cassettes, inserted in a head-to-tail orientation in a Sleeping Beauty DNA transposon vector, are efficiently inserted as a single genetic entity into the genome of cells of interest in a reaction catalyzed by the hyperactive SB100X transposase. The applicability of the sensor for screening purposes is demonstrated by the functional comparison of potent synthetic analogues of vitamin D3 designed for the treatment of psoriasis and cancer. In clones of human keratinocytes carrying from a single to numerous insertions of the vitamin D3 sensor, a sensitive sensor read-out is detected upon exposure to even low concentrations of vitamin D3 analogues. In comparative studies, the sensor unveils superior potency of new candidate drugs in comparison with analogues that are currently in clinical use.. Our findings demonstrate the use of the genetic sensor as a tool in first-line evaluation of new vitamin D3 analogues and pave the way for new types of drug delivery studies in sensor-transgenic animals.

Topics: Animals; Calcitriol; Cell Line; Cell Proliferation; Cell Survival; Cholecalciferol; DNA Transposable Elements; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Genetic Vectors; Green Fluorescent Proteins; Humans; Hydroxycholecalciferols; Keratinocytes; Microscopy, Fluorescence; Mutagenesis, Insertional; Response Elements; Retinoid X Receptors; Transfection; Transposases; Vitamin D-Binding Protein

Analogs of 1,25-dihydroxyvitamin D3 with a reversed configuration at C-1 or C-24 and E or Z geometry of the double bond at C-22 in the side chain or at C-5 in the triene system were examined for their antiproliferative activity in vitro against a spectrum of various human cancer cell lines. The analogs coded PRI-2201 (calcipotriol), PRI-2202 and PRI-2205, such as calcitriol and tacalcitol (used as a referential agents), revealed antiproliferative activity against human HL-60, HL-60/MX2, MCF-7, T47D, SCC-25 and mouse WEHI-3 cancer cell lines. The toxicity studies in vivo showed that PRI-2202 and PRI-2205 are less toxic than referential agents. Even at total doses of 2.5-5.0 mg/kg distributed during 5 successive days, no changes in body weight were observed. Calcitriol and tacalcitol showed toxicity in the same protocol at 100 times lower doses. Calcipotriol was lethal to all mice after administration of a total dose of 5.0 mg/kg. The analog PRI-2205 appeared to be more active in mouse Levis lung cancer tumor growth inhibition than calcitriol, calcipotriol or PRI-2202. This analog did not reveal calcemic activity at doses which inhibit tumor growth in vivo nor at higher doses.

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Calcitriol; Calcium; Carcinoma, Lewis Lung; CD11b Antigen; Cell Cycle; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Cholecalciferol; Coloring Agents; Female; Fibroblasts; HL-60 Cells; Humans; Lipopolysaccharide Receptors; Male; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Pancreatic Neoplasms; Prostatic Neoplasms; Rhodamines; Stereoisomerism; Tetrazolium Salts; Thiazoles

A variety of approaches (in vitro-/ex vivo studies, animal models, human studies and clinical trials) are available to assess compounds with potential antipsoriatic properties. Over the past few years various rodent models that mirror aspects of psoriasis phenotypes and/or pathogenesis have been created (e. g. knockout rodents, xenotransplantation models). Unfortunately these animal models do not reflect the complete pathogenesis of psoriasis. Therefore, screening procedures involving psoriatic lesions in humans are necessary. Even in the era of biologicals, the psoriasis plaque test (PPT) remains an important in vivo tool. In addition to screening potential antipsoriatic substances, the PPT can help answer other questions (frequency of use, dose-response relationship). A prerequisite for correct performance of PPT is knowledge of the toxicological and pharmacological data of the investigational compounds. The PPT is relatively simple, not time-consuming and allows the simultaneous testing of multiple substance. All the results from PPT must be confirmed by controlled clinical trials.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Biological Products; Calcineurin Inhibitors; Calcitriol; Cholecalciferol; Dermatologic Agents; Dose-Response Relationship, Drug; Drug Administration Schedule; Drug Evaluation, Preclinical; Humans; Psoriasis; Rodentia; Skin

The beta-catenin signaling pathway is deregulated in nearly all colon cancers. Nonhypercalcemic vitamin D3 (1alpha,25-dehydroxyvitamin D(3)) analogues are candidate drugs to treat this neoplasia. We show that these compounds promote the differentiation of human colon carcinoma SW480 cells expressing vitamin D receptors (VDRs) (SW480-ADH) but not that of a malignant subline (SW480-R) or metastasic derivative (SW620) cells lacking VDR. 1alpha,25(OH)2D(3) induced the expression of E-cadherin and other adhesion proteins (occludin, Zonula occludens [ZO]-1, ZO-2, vinculin) and promoted the translocation of beta-catenin, plakoglobin, and ZO-1 from the nucleus to the plasma membrane. Ligand-activated VDR competed with T cell transcription factor (TCF)-4 for beta-catenin binding. Accordingly, 1alpha,25(OH)2D(3) repressed beta-catenin-TCF-4 transcriptional activity. Moreover, VDR activity was enhanced by ectopic beta-catenin and reduced by TCF-4. Also, 1alpha,25(OH)2D(3) inhibited expression of beta-catenin-TCF-4-responsive genes, c-myc, peroxisome proliferator-activated receptor delta, Tcf-1, and CD44, whereas it induced expression of ZO-1. Our results show that 1alpha,25(OH)2D(3) induces E-cadherin and modulates beta-catenin-TCF-4 target genes in a manner opposite to that of beta-catenin, promoting the differentiation of colon carcinoma cells.

Topics: Active Transport, Cell Nucleus; Adenocarcinoma; Antineoplastic Agents; beta Catenin; Cadherins; Calcitriol; Cell Adhesion Molecules; Cell Differentiation; Cell Membrane; Cholecalciferol; Colonic Neoplasms; Cytoskeletal Proteins; Gene Expression Regulation, Neoplastic; Humans; Ligands; Macromolecular Substances; Phenotype; Protein Binding; Receptors, Calcitriol; RNA, Messenger; Signal Transduction; TCF Transcription Factors; Trans-Activators; Transcription Factor 7-Like 2 Protein; Transcription Factors; Transfection; Tumor Cells, Cultured; Vitamin D

The effects of cholesterol, 7-dehydrocholesterol, vitamin D3 and several synthetic vitamin D3 analogs on ribonuclease P (RNase P) were investigated using a cell-free system from the slime mold Dictyostelium discoideum. RNase P is an ubiquitous and essential enzyme that endonucleolytically cleaves all tRNA precursors to produce the mature 5' end. Among the compounds tested, only calcipotriol was capable of affecting RNase P activity, and revealed a bimodal action at the kinetic phase of the reaction. Depending on the concentration of the drug, both activation and inhibition of tRNA maturation were observed, indicating that calcipotriol may have a direct effect on tRNA biogenesis, possibly associated with the presence of a highly reactive small ring on the side chain of its molecule.

Topics: Animals; Calcitriol; Cholecalciferol; Cholesterol; Dehydrocholesterols; Dictyostelium; Dose-Response Relationship, Drug; Endoribonucleases; Enzyme Activation; Kinetics; Ribonuclease P; Ribonuclease, Pancreatic; RNA Processing, Post-Transcriptional; RNA, Catalytic; RNA, Transfer; Structure-Activity Relationship

In this paper, two methods are presented. One involves the separation of calcipotriol, a new synthetic analogue, from two related compounds, specifically cholecalciferol (Vitamin D3) and calcitriol (1,25-dihydroxyvitamin D3). The other involves the isolation and assay of calcipotriol from a topical ointment. The study was performed with reversed-phase high-performance liquid chromatography using an RP18 column and ultraviolet detection. Applying the method of Snyder, a mobile phase mixture containing methanol-acetonitrile-water (67:23:10, v/v) was found which achieved a total separation within 18 min. A mobile phase of methanol-water (80:20, v/v) attained a slower elution of calcipotriol. For isolation and assay of calcipotriol from an ointment (Daivonex), dissolution in chloroform gave the highest recovery (> 98%). The isolation and assay process can be performed within 2 h.

Topics: Acetonitriles; Calcitriol; Chloroform; Cholecalciferol; Chromatography, High Pressure Liquid; Methanol; Quality Control; Technology, Pharmaceutical; Time Factors; Water

Calcipotriol, 1,25(OH)2D3 and 1,24(OH)2D3 are potent drugs for the treatment of psoriasis. It has recently been published that these compounds induce epidermal hyperproliferation in hairless mouse skin.. The aim of our study was to examine the effect of vitamin D3 derivatives on epidermal growth, keratinization and permeability barrier function in vivo.. Calcipotriol, 1,25(OH)2D3 and 1,24(OH)2D3 in isopropanol or in an ointment formula were applied to normal hairless mouse skin. Transepidermal water loss (TEWL), a marker of cutaneous barrier function, and epidermal proliferation were determined at different time points 0-264 h after treatment. In addition, light and electron microscopy studies were performed.. A single treatment in solution led to a transient (2- to 3-fold) increase in TEWL after application of calcipotriol or 1,25(OH)2D3 and to a 3- to 6-fold increase in epidermal proliferation after application of each of the compounds. Repeated applications also resulted in an up to 3-fold increase in TEWL which persisted for 3 days after the end of the treatment. By light microscopy an increase in epidermal thickness was observed. There was no sign of inflammation. Electron microscopy studies showed the formation of a transitional cell zone as a sign of a premature keratinization.. These results demonstrate that in normal mouse skin vitamin D3 and its analogues disrupt the epidermal permeability barrier by induction of epidermal proliferation and premature keratinization but without morphological signs of inflammation.

Topics: 1-Propanol; Administration, Cutaneous; Animals; Calcitriol; Cell Division; Cholecalciferol; Dermatologic Agents; Dihydroxycholecalciferols; Epidermis; Keratins; Male; Mice; Mice, Hairless; Microscopy, Electron; Ointments; Permeability; Psoriasis; Skin; Time Factors; Water Loss, Insensible

Topics: Calcitriol; Cholecalciferol; Dermatologic Agents; Humans; Psoriasis

Starting with (20S)-20-(p-toluenesulfonyl)oxymethyl-pregna-1, 5-dien-3 alpha-ol (4), we synthesized three vitamin D analogs in 10 to 11 steps: 1 alpha, 26-dihydroxy-27-nor-vitamin D3 (1), its 3-epi analog (2), and 2 beta-methoxy-1 alpha, 26-dihydroxy-27-nor-vitamin D3 (3). We tested the derivatives in the murine mesenchymal cell line C3H1OT1/2. All substances were less potent in inhibition of cell proliferation, inhibition of adipocyte differentiation, and induction of gene activation, and had a lower affinity to the vitamin D receptor than the native vitamin D3 metabolite 1.25(OH)2D3. The affinity of 1 to the vitamin D binding protein was about three times higher than that of 1.25(OH)2D3.

Topics: Adipocytes; Animals; Calcitriol; Cell Differentiation; Cell Division; Chloramphenicol O-Acetyltransferase; Cholecalciferol; Fibroblasts; Genes, Reporter; Mice; Osteocalcin; Receptors, Calcitriol; Structure-Activity Relationship; Transcription, Genetic; Transfection; Vitamin D-Binding Protein

Although numerous studies have shown potent antiproliferative and differentiation-inducing effects of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) and its analogs on cells not directly related to bone metabolism, only few reports focussed on the effects of these analogs on bone. We compared the action of several recently developed analogs with that of 1,25-(OH)2D3 on human (MG-63) and rat (ROS 17/2.8) osteoblast-like cells and on in vitro bone resorption. In MG-63 cells the analogs EB1089 and KH1060 were about 166,000 and 14,000 times more potent than 1,25-(OH)2D3 in stimulating type I procollagen and 100 and 6,000 times more potent in stimulating osteocalcin production, respectively. Also in ROS 17/2.8 cells EB1089 and KH1060 were most potent in inducing osteocalcin synthesis. In vitro bone resorption was 2.3 and 17.5 times more potently stimulated by EB1089 and KH1060, respectively. In MG-63 cells, 1,25-(OH)2D3 and the analogs inhibited cell proliferation, whereas both 1,25-(OH)2D3 and the analogs stimulated the growth of ROS 17/2.8 cells. Differences in potency could neither be explained by affinity for the vitamin D receptor nor by a differential involvement of protein kinase C in the action of the analogs. Together, these data show that also in bone the analogs EB1089 and KH1060 are more potent than 1,25-(OH)2D3 but that the potency of the analogs compared to 1,25-(OH)2D3 is dependent on the biological response. On the basis of these observations it can be concluded that the reported reduced calcemic effect in vivo is not the result of a decreased responsiveness of bone to these analogs. Lastly, in view of eventual clinical application of 1,25-(OH)2D3-analogs, the observed stimulation of in vitro bone resorption and growth of an osteosarcoma cell line warrant in vivo studies to further examine these effects.

Topics: Animals; Antineoplastic Agents; Binding, Competitive; Bone Resorption; Calcitriol; Cell Division; Cells, Cultured; Cholecalciferol; Enzyme Inhibitors; Glyceryl Ethers; Humans; In Vitro Techniques; Osteoblasts; Osteocalcin; Osteosarcoma; Procollagen; Protein Kinase C; Rats; Receptors, Calcitriol; Structure-Activity Relationship; Tumor Cells, Cultured

Pro-inflammatory cytokines mediate their biological functions after they are secreted or released from intracellular to extracellular milieu. Keratinocytes have proven to be able to produce various cytokines including IL-1 and IL-8. Dysregulations of IL-1 and IL-8 were found in psoriatic lesions. Recently, vitamin D3 (VD3) was found to be an effective and safe therapy for psoriasis. In the present study, we investigated the effects of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and its analogue MC903 on IL-1 alpha and IL-8 secretion by human keratinocytes in vitro. Cultured normal human keratinocytes (NHKs) produced considerable amounts of IL-1 alpha but secreted less. In contrast, they produced less IL-8 and almost all molecules were secreted to the culture supernatants. Treatment of unstimulated NHKs with 1,25(OH)2D3 or MC903 showed little effects on IL-1 alpha production and secretion though they slightly enhanced IL-8. When NHKs were stimulated with tumour necrosis factor-alpha (TNF alpha), both IL-1 alpha and IL-8 secretions were enhanced and these enhancements were inhibited by 1,25(OH)2D3 or MC903. Stimulation of NHKs with phorbol 12-myristate 13-acetate(PMA) and lipopolysaccharide(LPS) resulted in an increase of IL-8 and decrease of IL-1 alpha in the culture supernatants. Addition of 1,25(OH)2D3 or MC903 inhibited the increased secretion of IL-8 but restored decreased secretion of IL-1 alpha from stimulated NHKs dose dependently. Hydrocortisone and cyclosporin A showed similar inhibitory effects on PMA/LPS-increased IL-8 secretion from NHKs but had little effect of restoring IL-1 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Calcitriol; Carcinoma, Squamous Cell; Cholecalciferol; Dermatologic Agents; Humans; Interleukin-1; Interleukin-8; Keratinocytes; Reference Values; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Vitamin D3, 1 alpha,25(OH)2D3, and its metabolites regulate the growth and differentiation of several cell types. Vitamin D3 and its analogue, calcipotriol (MC 903), inhibit the proliferation of cultured human and mouse keratinocytes and induce keratinocyte differentiation. Calcipotriol is effective in the treatment of psoriasis in which increased plasminogen activator activity has been reported. We analyzed therefore the effects of calcipotriol and vitamin D3 on the production of plasminogen activator (PA) activity in human keratinocytes and a mouse keratinocyte cell line. Caseinolysis-in-agarose assays indicated that vitamin D3 decreases total PA activity in both keratinocyte culture systems. Zymographic analyses of the medium indicated that the secreted activator was of the urokinase type (u-PA). A decrease was observed also in extracellular matrix and membrane-associated u-PA activity of vitamin D3 and calcipotriol treated cells. Immunoblotting analysis of the conditioned medium from human keratinocytes revealed a decrease in the u-PA protein levels. Accordingly, Northern hybridization analysis of the respective mRNAs indicated a rapid decrease in urokinase mRNA levels. Calcipotriol decreased u-PA activity also in the presence of inducers of u-PA activity like transforming growth factor-beta, epidermal growth factor, and phorbol-12-myristate-13-acetate. Calcipotriol also caused a decrease in tissue type PA (t-PA) activity of the keratinocytes. Most t-PA activity was associated with the extracellular matrices and cell membranes as revealed by zymographic analysis. Paradoxically, the secretion and deposition of the matrix of plasminogen activator inhibitor type 1 decreased in calcipotriol-treated cells. The results indicate that a major effect of vitamin D3 on cultured keratinocytes is a decrease of plasminogen activator activity.

Topics: Animals; Calcitriol; Cells, Cultured; Cholecalciferol; Dermatologic Agents; Extracellular Matrix; Humans; Keratinocytes; Mice; Mice, Inbred BALB C; Plasminogen Activator Inhibitor 1; Plasminogen Activators; Protein Biosynthesis; RNA, Messenger; Tetradecanoylphorbol Acetate; Transforming Growth Factor beta; Urokinase-Type Plasminogen Activator

The efficacy of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and the new analogue calcipotriene (MC 903) in the treatment of psoriasis was investigated. Eight patients with psoriasis were enrolled in a pilot study with systemically administered vitamin D3 and were given 1,25(OH)2D3 (Rocaltrol; Hoffmann-La Roche) in dosages from 0.5 microgram to 2 micrograms/day for a 6-month trial. In one patient, the psoriatic plaques resolved within 2 months after treatment (0.5 microgram/day) was initiated. Moderate improvement was noted in one other patient (1 microgram/day). The serum level of 1,25(OH)2D before treatment was not less than the normal range of the adult population. Side effects of systemically administered 1,25(OH)2D3 included a dose-dependent increase in the 24-hour urinary calcium excretion and a decrease in the total number of platelets. Seven patients with symmetric plaque-type psoriasis were treated topically with 2 micrograms/g of 1,25(OH)2D3 in petrolatum. During 3 months of follow-up, mild improvement was noted in three patients. Five patients in the calcipotriene study were part of a nationwide double-blind placebo-controlled trial by Bristol-Myers Squibb. Moderate to marked improvement was noted in the two patients who received 50 micrograms/g of calcipotriene topically. The three patients who received placebo showed no response. We conclude that a subset of patients with psoriasis responds well to 1,25(OH)2D3. Calcipotriene is efficacious and an excellent alternative to topically applied corticosteroids.

Topics: Administration, Oral; Administration, Topical; Adult; Aged; Calcitriol; Cholecalciferol; Dermatologic Agents; Female; Humans; Male; Middle Aged; Pilot Projects; Psoriasis

The hormonally active form of vitamin D3, 1,25-dihydroxy vitamin D3 [1,25-(OH)2-D3; calcitriol], regulates the differentiation and proliferation of epidermal keratinocytes in vitro. MC 903 (calcipotriol) is a novel vitamin D3 analogue which is at least 100 times less potent than 1,25-(OH)-2-D3 in its effects on calcium homeostasis. The present study compared the effects of MC 903 and 1,25-(OH)2-D3 on terminal differentiation and proliferation of cultured normal human keratinocytes. Keratinocytes were grown in McCoy's 5A medium supplemented with penicillin (50 IU/ml), streptomycin (50 micrograms/ml), L-serine (4 X 10(-4) M), and 10% human type AB serum. MC 903, 1,25-(OH)2-D3 or 1 alpha-OH-D3 (10(-12) M--10(-8) M) was added with each feeding when cultures became preconfluent. After incubation for 24 h with D3 vitamins, cultures were extracted for transglutaminase, and the enzyme activity was indexed against DNA content. The activity of transglutaminase, the enzyme responsible for cross-linking the proteins of the cornified envelope, was maximally stimulated by 388% with MC 903 (10(-8) M), by 328% with 1,25-(OH)2-D3 (10(-8) M), and by 27% with 1 alpha-OH-D3 (10(-8) M) compared with vehicle. After incubation for 2 weeks the number of keratinocytes with cornified envelopes had increased by 288% with MC 903 (10(-8) M), by 360% with 1,25-(OH)2-D3 (10(-8) M), and by 149% with 1 alpha-OH-D3 (10(-8) M) compared with vehicle.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Calcitriol; Cell Differentiation; Cell Division; Cells, Cultured; Cholecalciferol; Humans; Keratinocytes