VDR/vitamin D receptor.

在没有 IL-17A 或 IL-22 刺激的情况下，卡泊三醇对 NHEK 细胞中 IL-8 mRNA 的表达没有影响 (2-20 nM) 或仅略微增加 (0.2 nM)。我们早期的研究通过添加IL-17A和IL-22得到了验证，这极大地提高了IL-8的mRNA表达。以剂量依赖性方式，2、20 和 40 nM 卡泊三醇可以抑制这种增加的 IL-8 mRNA 表达 [1]。当药物作用于自然杀伤 (NK) 细胞时，NK 细胞毒性受体 (KIR) 的表达会受到调节。将卡泊三醇、FTY720 或 100、10 或 1 ng/mL 1,25(OH)2D3 作为人类 NK 细胞的预处理，持续 4 小时。经过 4 小时的潜伏期后，所有三种剂量的 1,25(OH)2D3、卡泊三醇和 FTY720 均显着提高了 NK 细胞表面 NKp30 的表达 [2]。

Calcipotriol (MC903)的局部应用诱发特应性皮炎/AD （Atopic Dermatitis） | Basic Protocol: TOPICAL APPLICATION OF Calcipotriol (MC903) INDUCES AD-LIKE SKIN INFLAMMATION[6]
材料
小鼠（C57BL/6J野生型，6-10周龄）
异氟烷
Calcipotriol (MC903)，钙泊三醇
乙醇（EtOH），纯，190度
37°C鼠标加热垫
精密刻度盘测厚仪
高精度实验室天秤
支持Protocol 1-5的其他试剂和设备。
第0-14天：诱发皮炎
1.使用2L/min氧气中的4%异氟烷进行诱导麻醉，使用0.4L/min氧气中2%异氟烷进行维持麻醉。
2.使用高精度实验室天平称量小鼠体重，并将动物放在37°C的小鼠加热垫上以保持体温。记录治疗过程中的体重。
3.在轻度麻醉下，使用精密厚度表/precision-dial thickness gauge测量左耳和右耳厚度。
4.根据治疗方案，在每个治疗日，使用2-20µl移液管将1 nM Calcipotriol (MC903)（溶解在100%无水乙醇/EtOH中），局部涂抹在每只耳朵的两侧（耳朵背侧和腹侧各10μl，每只耳朵总体积为20μl）。对照组小鼠仅用相同体积的乙醇（作为空白对照）平行治疗。
5.治疗后，将动物放在37°C的小鼠加热垫上，以在恢复期间保持恒定的体温，然后回到笼子里。
6.在接下来的2周内（参见治疗方案），在局部应用Calcipotriol (MC903)或乙醇之前重复测量（步骤2-5）。总共将使用7次Calcipotriol (MC903)或乙醇，最后一次局部使用将在第14天进行。
第14天：瘙痒评估
7.在实验结束前24小时的第14天，记录小鼠的视频，并通过延时录像量化瘙痒事件。确定并量化30分钟的瘙痒事件。
第15天：实验终点
8.在实验终点（第15天），通过二氧化碳窒息对小鼠实施安乐死，重复测量（步骤3），并使用无菌尖剪刀和镊子采集耳皮组织（Alam等人，2020，Mac Daniel，Buckwalter，Gueirard，&Ménard，2016）。对于耳引流淋巴结，仔细切割相应颈部区域的皮肤并解剖耳淋巴结（Alam等人，2020，Mac Daniel等人，2016）。

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双氯芬酸加 DFMO 加卡泊三醇组导致每 32 只动物中有 1 只死亡，而该组中的所有其他动物均存活。每个组的存活率相似。与安慰剂相比（线性回归模型），双氯芬酸加卡泊三醇 (p=0.018) 和双氯芬酸加 DFMO 加卡泊三醇 (p=0.002) 治疗组的体重增加显着减少 [3]。

白细胞介素-17A和-22参与银屑病的发病。Cathelicidin LL37不仅作为抗菌肽，还作为自身炎症介质。1,25-二羟基维生素D3类似物，如钙泊三醇，用作银屑病的局部治疗。然而，钙泊三醇对IL-17A/IL-22刺激的角质形成细胞表达/产生人组织蛋白酶抗菌蛋白（hCAP18）和LL37肽的影响仍然存在争议。为了评估钙泊三醇对hCAP18和LL37产生的调节作用，我们通过实时qPCR、ELISA、蛋白质印迹和免疫细胞染色分析了IL-17A/IL-22刺激的培养的人角质形成细胞中hCAP18 mRNA的表达和hCAP18/LL37肽的产生。通过蛋白质印迹法，在用IL-17/IL-22培养72小时的角质形成细胞中检测到hCAP18蛋白。钙泊三醇增加了IL-17/IL-22刺激的角质形成细胞中hCAP18 mRNA的表达。然而，钙泊三醇降低了培养上清液中的LL37肽。免疫染色显示，过量产生的LL37存在于细胞内。LL37通过与细胞外DNA的相互作用促进银屑病，但可能通过干扰细胞质DNA抑制银屑病。[1]

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在这项研究中，研究人员在这里描述了三种药物的效果，这些药物要么被批准，要么有可能通过人类自然杀伤细胞（NK）和树突状细胞（DC）的体外活性治疗多发性硬化症（MS）患者。我们的结果表明，维生素D3、钙泊三醇和FTY720的生物活性代谢产物1,25（OH）2D3可增强IL-2激活的K562和RAJI肿瘤细胞系以及未成熟（i）和成熟（m）DCs的NK细胞裂解，其效果各不相同。这些结果与药物上调NK细胞表面NK细胞毒性受体NKp30和NKp44以及NKG2D表达的能力相证实。此外，它们还能下调杀伤抑制受体CD158的表达。这三种药物下调iDCs表面CCR6的表达，而维生素D3和钙泊三醇倾向于上调mDCs上CCR7的表达，这表明它们可能会影响DCs向淋巴结的迁移。最后，维生素D3、钙泊三醇和FTY720增强K562细胞的NK17/NK1细胞裂解，表明这些药物的可能作用机制是通过激活这些新描述的细胞。总之，我们的研究结果显示了维生素D3、钙泊三醇和FTY720对先天免疫系统细胞的新作用机制。[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

60.69

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

注意： (1). 本产品在运输和储存过程中需避光。  (2). 请将本产品存放在密封且受保护的环境中（例如氮气保护），避免吸湿/受潮。  (3). 该产品在溶液状态不稳定，请现配现用。

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.06 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (6.06 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (6.06 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: ≥ 2.5 mg/mL (6.06 mM) (饱和度未知) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
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10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
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