VDR/vitamin D receptor.

In the absence of IL-17A or IL-22 stimulation, calcipotriol either had no effect (2–20 nM) or only slightly increased (0.2 nM) the expression of IL-8 mRNA in NHEK cells. Our earlier research was validated by the addition of IL-17A and IL-22, which greatly raised the mRNA expression of IL-8. In a dose-dependent manner, 2, 20, and 40 nM calcipotriol can suppress this increased IL-8 mRNA expression [1]. The expression of NK cytotoxic receptors, or KIRs, is modulated when medicines are administered to natural killer (NK) cells. For four hours, calcipotriol, FTY720, or 100, 10, or 1 ng/mL of 1,25(OH)2D3 were applied as a pretreatment to human NK cells. Following a 4-hour incubation period, the expression of NKp30 on the surface of NK cells was considerably elevated by all three doses of 1,25(OH)2D3, calcipotriol, and FTY720 [2].

Basic Protocol: TOPICAL APPLICATION OF MC903 INDUCES AD-LIKE SKIN INFLAMMATION[6]
Materials
Mice (C57BL/6J wild type, 6-10 weeks old)
Isoflurane (Baxter, cat. no. 100190360400)
MC903, calcipotriol, hydrate
Ethyl alcohol (EtOH, ethanol) , pure, 190 proof
37°C mouse heating pad
Precision-dial thickness gauge
High-precision laboratory scale
Additional reagents and equipment for Support Protocols 1–5.
Days 0-14: Induce dermatitis
1. Anesthetize mice using 4% isoflurane in 2 L/min of oxygen for induction and 2% isoflurane in 0.4 L/min of oxygen for maintenance.
2. Weigh mice using a high-precision laboratory scale and place animals on a 37°C mouse heating pad to maintain body temperature. Record weights over the course of treatment.
3. Measure both left and right ear thickness using a precision-dial thickness gauge under light anesthesia.
4. Topically, apply 1 nM MC903 in 100% absolute ethyl alcohol (EtOH) using a 2-20 µl pipette to both sides of each ear (10 μl on dorsal and ventral ear sides for a total volume of 20 μl per ear) on every treatment day according to the treatment regimen. Control mice are treated in parallel with the same volume of EtOH (as a vehicle control) only.
5. After the treatment, keep animals on a 37°C mouse heating pad to maintain constant body temperature during recovery, then return to cages.
6. Over the next 2 weeks (see treatment regimen), repeat measurements before topical application of MC903 or EtOH (steps 2-5). A total of 7 applications of MC903 or EtOH will be applied, with the final topical application on day 14.
Day 14: Itch assessment
7. On day 14, 24 hr before the experimental endpoint, record a video of the mice and quantify itch events via time-lapse videography. Determine and quantify itch events for 30 min.
Day 15: Experimental endpoint
8. At the experimental endpoint (day 15), euthanize mice by CO2 asphyxiation, repeat measurements (step 3), and harvest ear skin tissues using sterile sharp scissors and forceps (Alam et al., 2020, Mac-Daniel, Buckwalter, Gueirard, & Ménard, 2016). For ear draining lymph nodes, carefully cut the skin at the corresponding neck region and dissect the auricular lymph nodes (Alam et al., 2020, Mac-Daniel et al., 2016)

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The diclofenac plus DFMO plus calcipotriol group resulted in 1 out of every 32 animals dying, whereas all other animals in this group survived. Every group had a similar survival rate. Diclofenac plus calcipotriol (p=0.018) and diclofenac plus DFMO plus calcipotriol (p=0.002) therapy groups showed considerably less weight gain when compared to the placebo (linear regression model) [3].

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Interleukins (IL)-17A and -22 are involved in the patho-genesis of psoriasis. Cathelicidin LL37 serves as not only antimicrobial peptide but also as autoinflammatory mediator. 1,25-Dihydroxyvitamin D3 analogues, such as calcipotriol, are used as topical treatment for psoriasis. However, the effect of calcipotriol on the mRNA expression/production of human cathelicidin antimicrobial protein (hCAP18) and LL37 peptide by IL-17A/IL-22-stimulated keratinocytes remains controversial. To evaluate the modulatory action of calcipotriol on the production of hCAP18 and LL37, we analysed hCAP18 mRNA expression and hCAP18/LL37 peptide production in IL-17A/IL-22-stimulated cultured human keratinocytes by real-time qPCR, ELISA, western blotting, and immunocytostaining. By western blotting, hCAP18 protein was detected in keratinocytes cultured for 72 h with IL-17/IL-22. Calcipotriol increased hCAP18 mRNA expression in IL-17/IL-22-stimulated keratinocytes. However, LL37 peptide in the culture supernatants was reduced by calcipotriol. Immunostaining revealed that the overproduced LL37 resides within the cells. LL37 promotes psoriasis via interaction with extracellular DNA, but may suppress psoriasis by interfering cytosolic DNA.[1]

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In this study, researchers 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.[2]

A total of 160 SKH-1 mice were randomized to one placebo group and four chemoprevention groups (diclofenac plus difluoromethylornithine; diclofenac plus calcipotriol; difluoromethylornithine plus calcitriol; and diclofenac plus difluoromethylornithine plus calcipotriol). The mice received UVB radiation for 20 weeks followed by 17 weeks with topical application of chemoprevention. The number of mice with tumors, number of tumors per group and tumor area size were compared using a linear regression model. Results: Chemoprevention with diclonefac plus calcipotriol and diclonefac plus difluoromethylornithine had a significant inhibiting effect on the number of tumors per group and the area of tumors. Moreover, diclonefac plus difluoromethylornithine had a significant inhibiting effect on the number of mice with tumors. Conclusion: Potentially, non-melanoma skin cancer in humans may be prevented with these agents with few adverse effects. Therefore, clinical studies are needed to determine their therapeutic/preventive effect and possible adverse effects.[3]

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Calcipotriol (MC903) can be used to induce Atopic Dermatitis (AD) mice model
Calcipotriol (MC903) was used to establish AD-like mice model. A murine model of AD-like disease was established as in a prior study. Briefly, mice were administered a daily topical dose of a 15 µL 0.005% Calcipotriol (MC903) scalp solution (MC903), which was applied to the dorsal and ventral sides of each ear for 12 consecutive days. Control animals received 15 µL of ethanol instead. Mice in the Calcipotriol (MC903) + poly (I:C) group were then treated with poly (I:C) in a concentration of 5 µg/g bodyweight. The impact of poly (I:C) treatment on these animals was assessed based upon changes in lesions, bodyweight, ear thickness, and histopathological findings. In addition, serum interleukin 4 (IL-4), interferon-γ (IFN-γ), immunoglobulin E (IgE), IL-13, and TSLP levels were measured using enzyme-linked immunosorbent assay (ELISA), while tissue IL-13 and TSLP levels were assessed using ELISA, Western blotting, and immunohistochemical staining, and mast cell infiltration was assessed through toluidine blue (TBO) staining.[from: Ann Transl Med. 2022 Feb;10(4):209. ]

Absorption, Distribution and Excretion
Clinical studies with radiolabeled ointment indicate that approximately 6% (+3%, SD) of the applied dose of calcipotriene is absorbed systemically when the ointment is applied topically to psoriasis plaques or 5% (+2.6%, SO) when applied to normal skin.
The active form of the vitamin, 1,25-dihydroxy vitamin D3 (calcitriol), is known to be recycled via the liver and excreted in the bile. There is evidence that maternal 1,25-dihydroxy vitamin D3 (calcitriol) may enter the fetal circulation, but it is not known whether it is excreted in human milk.

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Metabolism / Metabolites
Hepatic. Calcipotriene metabolism following systemic uptake is rapid, and occurs via a similar pathway to the natural hormone. The primary metabolites are much less potent than the parent compound.
Hepatic. Calcipotriene metabolism following systemic uptake is rapid, and occurs via a similar pathway to the natural hormone. The primary metabolites are much less potent than the parent compound.
Route of Elimination: The active form of the vitamin, 1,25-dihydroxy vitamin D3 (calcitriol), is known to be recycled via the liver and excreted in the bile. There is evidence that maternal 1,25-dihydroxy vitamin D3 (calcitriol) may enter the fetal circulation, but it is not known whether it is excreted in human milk.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the use of calcipotriene during breastfeeding. Because it is poorly absorbed after topical application, calcipotriene is probably a low risk to the nursing infant and is generally considered acceptable during breastfeeding, although some sources recommend avoiding the nipple area. Avoid application of the combination products containing betamethasone (Enstilar) to the breast. Only water-miscible cream or gel products should be applied to the breast because ointments may expose the infant to high levels of mineral paraffins via licking.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

[1]. Calcipotriol Increases hCAP18 mRNA Expression but Inhibits Extracellular LL37 Peptide Production in IL-17/IL-22-stimulated Normal Human Epidermal Keratinocytes. Acta Derm Venereol. 2014 Sep;94(5):512-6.

[2]. Effects of vitamin D3, calcipotriol and FTY720 on the expression of surface molecules and cytolytic activities of human natural killer cells and dendritic cells. Toxins (Basel). 2013 Oct 28;5(11):1932-47.

[3]. Combination chemoprevention with diclofenac, calcipotriol and difluoromethylornithine inhibits development of non-melanoma skin cancer in mice. Anticancer Res. 2013 Aug;33(8):3033-9.

[4]. Induction of Antigen-Specific Tolerance by Peripheral Phagocytosis. US 20150283231 A1.

[5]. Nanoemulsion loaded gel for topical co-delivery of clobitasol propionate and calcipotriol in psoriasis. Nanomedicine. 2017 May;13(4):1473-1482.

[6]. A Mouse Model of MC903-Induced Atopic Dermatitis. Atopic Dermatitis. Curr Protoc. 2023 Mar;3(3):e695.

Calcipotriol is a seco-cholestane that is 26,27-cyclo-9,10-secocholesta-5,7,10,22-tetraene carrying additional hydroxy substituents at positions 1, 3 and 24. It is used (as its hydrate) in combination with betamethasone dipropionate, a corticosteroid, for the topical treatment of plaque psoriasis in adult patients. It has a role as a drug allergen and an antipsoriatic. It is a member of cyclopropanes, a secondary alcohol, a triol, a hydroxy seco-steroid and a seco-cholestane.
Calcipotriol (INN) or calcipotriene (USAN) is a sythetic derivative of calcitriol or Vitamin D.
Calcipotriene is a Vitamin D Analog.
Calcipotriene is a synthetic vitamin D derivative usually formulated for topical dermatological use, antipsoriatic Calcipotriene (calcipotriol) competes equally with active 1,25-hydroxy-2D3 (the natural form of vitamin D) for 1,25-hydroxy-2D3 receptors in regulating cell proliferation and differentiation. It induces differentiation and suppresses proliferation of keratinocytes, reversing abnormal keratinocyte changes in psoriasis, and leads to normalization of epidermal growth. (NCI04)
Calcipotriol is only found in individuals that have used or taken this drug. It is a synthetic derivative of calcitriol or Vitamin D.The precise mechanism of calcipotriol in remitting psoriasis is not well-understood. However, it has been shown to have comparable affinity with calcitriol for the Vitamin D receptor, while being less than 1% as active as the calcitriol in regulating calcium metabolism. The Vitamin D receptor (VDR) belongs to the steroid/thyroid receptor superfamily, and is found on the cells of many different tissues including the thyroid, bone, kindney, and T cells of the immune system. T cells are known to play a role in psoriasis, and it is thought that the binding of calcipotriol to the VDR modulates the T cells gene transcription of cell differentiation and proliferation related genes.
See also: Calcipotriene hydrate (active moiety of); Betamethasone Dipropionate; Calcipotriene (component of); Calcipotriene; niacinamide (component of) ... View More ...

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Drug Indication
For the treatment of moderate plaque psoriasis in adults.
Treatment of psoriasis

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Mechanism of Action
The precise mechanism of calcipotriol in remitting psoriasis is not well-understood, however, it has been shown to have comparable affinity with calcitriol for the Vitamin D receptor while being less than 1% the activity in regulating calcium metabolism. The Vitamin D receptor (VDR) belongs to the steroid/thyroid receptor superfamily, and is found on the cells of many different tissues including the thyroid, bone, kindney, and T cells of the immune system. T cells are known to play a role in psoriasis and are believed to undergo modulation of gene expression with binding of calcipotriol to the VDR. This modulation is thought to affect gene products related to cell differentiation and proliferation.

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Calcipotriol (MC903) may be used to induce Atopic Dermatitis (AD) mice model
Atopic dermatitis (AD) is a chronic, relapsing, and extremely pruritic inflammatory skin disease with a particular impact on children. AD pathogenesis is not yet fully understood, and there is no curative treatment for this disease. Therefore, several genetically or chemically-induced AD mouse models have been developed. These preclinical mouse models are an indispensable research tool for studying AD pathogenesis and evaluating the efficacy of new candidate AD therapeutics. A commonly used mouse model of AD has been developed using the topical application of a low-calcemic analog of vitamin D3, MC903, to induce AD-like inflammatory phenotypes that closely resemble human AD. Moreover, this model shows a minimal effect on systemic calcium metabolism that is observed in the vitamin D3-induced AD model. Thus, an expanding number of studies use the MC903-induced AD model to interrogate AD pathobiology in vivo and to test new candidate small molecule and monoclonal antibody therapies. This protocol describes in detail functional measurements including the measurement of skin thickness, which is a surrogate marker for ear skin inflammation, as well as itch assessment, histological evaluation to assess the structural changes associated with AD skin inflammation, and preparation of single-cell suspensions from ear skin and draining lymph nodes for the assessment of inflammatory leukocyte subset infiltration in these tissues using flow cytometry. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Topical application of MC903 induces AD-like skin inflammation Support Protocol 1: Measurement of ear skin thickness Support Protocol 2: Itch assessment Support Protocol 3: Dissection of ear skin and ear draining lymph nodes Support Protocol 4: Histological evaluation and quantification Support Protocol 5: Preparation of single-cell suspension from ear skin and draining lymph nodes for the assessment of inflammatory immune cell infiltration using flow cytometry. [Reference: A Mouse Model of MC903-Induced Atopic Dermatitis. Curr Protoc. 2023 Mar;3(3):e695.  doi: 10.1002/cpz1.695.]

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Relative to vehicle control treatment, poly (I:C) administration was associated with a significant exacerbation of calcipotriol-induced AD-like murine skin lesions. In animals treated with poly (I:C), the levels of serum IL-4, IL-13 and TSLP increased significantly, while the level of IFN-γ did not change. It also increased IL-13 and TSLP levels in skin lesions relative to the control-group mice and increased dermal mast cell infiltration and IgE production. Conclusions These data indicate that poly (I:C) treatment and exogenous activation of TLR3 exacerbate murine calcipotriol-induced AD-like skin lesions in part by increasing the production of TSLP and other T-helper 2 (Th2)-related cytokines.[Reference: Polyinosinic:polycytidylic acid aggravates calcipotriol-induced atopic dermatitis-like skin lesions in mice by increasing the expression of thymic stromal lymphopoietin. Ann Transl Med. 2022 Feb;10(4):209.]

C27H40O3

412.61

412.297

, 78.60; H, 9.77; O, 11.63

112965-21-6

Calcipotriol monohydrate;147657-22-5;Impurity F of Calcipotriol;112875-61-3

5288783

White to off-white solid powder

1.1±0.1 g/cm3

582.0±50.0 °C at 760 mmHg

166-168ºC

250.6±24.7 °C

0.0±3.7 mmHg at 25°C

1.580

5.43

3

3

5

30

743

7

C[C@H](/C=C/[C@H](C1CC1)O)[C@H]2CC[C@@H]\3[C@@]2(CCC/C3=C\C=C/4\C[C@H](C[C@@H](C4=C)O)O)C

LWQQLNNNIPYSNX-JQWURIRRSA-N

InChI=1S/C27H40O3/c1-17(6-13-25(29)20-8-9-20)23-11-12-24-19(5-4-14-27(23,24)3)7-10-21-15-22(28)16-26(30)18(21)2/h6-7,10,13,17,20,22-26,28-30H,2,4-5,8-9,11-12,14-16H2,1,3H3/b13-6+,19-7+,21-10-/t17-,22+,23-,24+,25-,26-,27-/m1/s1

(5Z,7E,22E,24S)-24-Cyclopropyl-9,10-secochola-5,7,10(19),22-tetraene-1alpha,3beta,24-triol

Calcipotriene; calcipotriol; MC-903PRI; 2201MC903PRI-2201; Calcitrene ; CCRIS 7700; Daivonex; Dovonex; MC 903; Psorcutan Sorilux.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.  (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.  (3). This product is not stable in solution, please use freshly prepared working solution for optimal results.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~242.37 mM)
Ethanol : ~50 mg/mL (~121.18 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.06 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (6.06 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (6.06 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: ≥ 2.5 mg/mL (6.06 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)