FTY720 (S)-磷酸盐是一种 S1PR1 激动剂。 FTY720 (S)-磷酸盐（Tys，1 µM）不会影响 S1PR1 泛素化的诱导，但它确实维持 S1PR1 蛋白的表达并通过 S1PR1 改善人肺动脉内皮细胞屏障。此外，FTY720 (S)-磷酸盐 (0.01–50 µM) 会减少 β-抑制蛋白向 S1PR1 的招募[1]。

在患有博来霉素诱导的急性肺损伤 (ALI) 的小鼠中，FTY720 (S)-磷酸盐（0.5 mg/kg，腹腔注射）可减少肺组织白细胞浸润并维持肺部 S1PR1 表达[1]。

β-抑制蛋白激活[1]
使用Invitrogen Tango™EDG1-bla U2OS细胞检测法测定β-arrestin活化。简而言之，将EDG1 bla U2OS细胞在FreeStyle™表达培养基中生长48小时。然后，加入1µM的S1P、Tys、1R、FTY720、p-FTY720或10µM的SEW，并将细胞在37°C/5%CO2的加湿培养箱中培养5小时。加入荧光底物，将细胞在室温下避光2小时。检测荧光强度，计算每个孔的蓝/绿发射比，并将其用作S1PR1激活指示剂。实验重复3次，最终比率表示为平均值±S.E.M。

Bleomycin ALI models [1]
Male C57BL/6 (20–25 g) mice 8–10 weeks old received a single intratracheal dose of bleomycin at 0.6 U/kg (or sterile saline) on Day 0 followed immediately by intraperitoneal injection of Tys (0.5 mg/kg), FTY720 (0.5 mg/kg), or saline. Additional doses of Tys or FTY720 were injected on Days 3 and 6. Bronchoalveolar lavage (BAL) fluid and lungs were then collected on Day 7. BAL fluid was used to detect BAL protein levels, WBC count, and WBC differential count. Lungs were perfused with saline to remove blood for Western blot, tissue albumin, and histopathology evaluation. Peripheral blood was obtained on Day 7 for examination of total cell counts and lymphocytes. Experiments were repeated 3 times. 6–10 mice were used per experimental group.

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To block egress of lymphocytes, 9-week-old female NOD mice were either given 1.5 mg/kg FTY720 in sterile water or equal volume of sterile water by gavage daily for a week. Next the mice were sacrificed and immune cell populations were detected in MLN, PLN, and spleen by flow cytometry.[2]

[1]. FTY720 (s)-phosphonate preserves sphingosine 1-phosphate receptor 1 expression and exhibits superior barrier protection to FTY720 in acute lung injury. Crit Care Med. 2014 Mar;42(3):e189-99.

[2]. Clostridium butyricum CGMCC0313.1 Protects against Autoimmune Diabetes by Modulating Intestinal Immune Homeostasis and Inducing Pancreatic Regulatory T Cells. Front Immunol. 2017 Oct 19;8:1345.

Objective: Effective therapies are needed to reverse the increased vascular permeability that characterizes acute inflammatory diseases such as acute lung injury. FTY720 is a pharmaceutical analog of the potent barrier-enhancing phospholipid, sphingosine 1-phosphate. Because both FTY720 and sphingosine 1-phosphate have properties that may limit their usefulness in patients with acute lung injury, alternative compounds are needed for therapeutic use. The objective of this study is to characterize the effects of FTY720 (S)-phosphonate, a novel analog of FTY720-phosphate, on variables of pulmonary vascular permeability in vitro and alveolar-capillary permeability in vivo. Setting: University-affiliated research institute. Subjects: Cultured human pulmonary endothelial cells; C57BL/6 mice. Interventions: Endothelial cells were stimulated with sphingosine 1-phosphate receptor 1 agonists to determine effects on sphingosine 1-phosphate receptor 1 expression. Acute lung injury was induced in C57BL/6 mice with bleomycin to assess effects of sphingosine 1-phosphate receptor 1 agonists. Measurements and main results: FTY720 (S)-phosphonate potently increases human pulmonary endothelial cell barrier function in vitro as measured by transendothelial electrical resistance. Reduction of sphingosine 1-phosphate receptor 1 with small interference RNA significantly attenuates this transendothelial electrical resistance elevation. FTY720 (S)-phosphonate maintains endothelial sphingosine 1-phosphate receptor 1 protein expression in contrast to greater than 50% reduction after incubation with sphingosine 1-phosphate, FTY720, or other sphingosine 1-phosphate receptor 1 agonists. FTY720 (S)-phosphonate does not induce β-arrestin recruitment, sphingosine 1-phosphate receptor 1 ubiquitination, and proteosomal degradation that occur after other agonists. Intraperitoneal administration of FTY720 (S)-phosphonate every other day for 1 week in normal or bleomycin-injured mice maintains significantly higher lung sphingosine 1-phosphate receptor 1 expression compared with FTY720. FTY720 fails to protect against bleomycin-induced acute lung injury in mice, while FTY720 (S)-phosphonate significantly decreases lung leak and inflammation. Conclusion: FTY720 (S)-phosphonate is a promising barrier-promoting agent that effectively maintains sphingosine 1-phosphate receptor 1 levels and improves outcomes in the bleomycin model of acute lung injury. [1]

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FTY720 Suppresses the Accumulation of α4β7+ Tregs but Not the Total Tregs in the PLN To confirm if CB0313.1 promoted the migration of gut-primed Tregs to PLN, we administrated NOD mice with FTY720 by gavage, which inhibits T cells circulation and traps them in the lymph node (36). Indeed, FTY720 resulted in a decreased proportion of splenic Tregs (Figure 7A), whereas an increased fraction of Tregs in the MLN compared to the NOD-CB mice (Figure 7B). FTY720 inhibited the migration of α4β7+ Tregs away from MLN (Figure 7C). In the PLN, FTY720 suppressed the accumulation of α4β7+ Tregs (Figure 7E), however, did not significantly change the total proportion of Tregs (Figure 7D), suggesting that the majority of Tregs in the PLN might be induced locally and migrated α4β7+ Tregs represented only a small fraction (Figure S6 in Supplementary Material).[2]

C19H34NO5P

387.45

387.217

C, 58.90; H, 8.85; N, 3.62; O, 20.65; P, 7.99

402616-26-6

(S)-FTY720-phosphonate;1142015-10-8;Fingolimod phosphate;402615-91-2

11452022

White to off-white solid powder

1.2±0.1 g/cm3

584.2±60.0 °C at 760 mmHg

184-186ºC

307.1±32.9 °C

0.0±1.7 mmHg at 25°C

1.541

4.27

122.82

4

6

14

26

409

1

OC[C@](CCC1=CC=C(CCCCCCCC)C=C1)(N)COP(O)(O)=O

LRFKWQGGENFBFO-IBGZPJMESA-N

InChI=1S/C19H34NO5P/c1-2-3-4-5-6-7-8-17-9-11-18(12-10-17)13-14-19(20,15-21)16-25-26(22,23)24/h9-12,21H,2-8,13-16,20H2,1H3,(H2,22,23,24)/t19-/m0/s1

[(2S)-2-amino-2-(hydroxymethyl)-4-(4-octylphenyl)butyl] dihydrogen phosphate

(S)FTY720P S-FTY720P(S)-FTY720P (S) FTY720P FTY-720 (S)-Phosphate(S)-FTY720 phosphate FTY720 (S)-Phosphate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 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 : ~1 mg/mL (~2.58 mM)
H2O : < 0.1 mg/mL

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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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：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
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；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。