S1P1 ( IC50 = 1.88 nM )

Etrasimod arginine/APD-334 是一种结构新颖、选择性、功能性的 S1P1 拮抗剂。在表达 HA 标记的 S1P1 的 CHO 细胞中，APD334 的 IC50 值为 1.88 nM。观察到对人 S1P4 和 S1P5 的中等激动作用，但在效力和功效方面相对于 S1P1 有所降低。 APD334 对人 S1P2 和 S1P3 没有任何激动或拮抗作用。 APD334 口服给药后实现了良好的中枢暴露，并在多个临床前物种中具有良好的药代动力学特征。 S1P1 活性在小鼠 (EC50=0.44 nM)、大鼠 (EC50=0.32 nM)、狗 (EC50=0.34 nM) 和猴子 (EC50=0.32 nM) 中保持不变[1]。

Etrasimod 精氨酸/APD-334 在所有物种中具有相对较低的全身清除率（<肝血流量的 4%）和较高的 Cmax。与啮齿动物相比，狗和猴子的分布容积 (Vss) 显着减少。口服生物利用度在 40-100% 范围内，终末期半衰期从猴子的 6 小时到狗的长达 29 小时不等。 siponimod（另一种目前正在进行人体试验的 S1P1 调节剂）的大鼠和猴子 t1/2 值已公开，分别为 6 小时和 19 小时[1]。大鼠：在雄性 Sprague-Dawley 大鼠中测定了 APD334 对血液淋巴细胞减少症的诱导作用。简言之，雄性大鼠口服0.5%甲基纤维素(MC)水溶液中配制的APD334，口服剂量为0(仅载体)、0.03(仅小鼠)、0.1、0.3或1mg/kg。在给药后 0、1、3、5、8、16、24、32、48 和 72 小时收集大鼠血液样本[1]。小鼠：在雄性 BALB/c 小鼠中测定了 APD334 对血液淋巴细胞减少症的诱导作用。简言之，向雄性小鼠给予0(仅载体)、0.03(仅小鼠)、0.1、0.3或1mg/kg口服剂量的在0.5％甲基纤维素(MC)水溶液中配制的APD334。在给药后 0、1、3、5、8、16、24 和 32 小时采集小鼠血液样本[1]。

Rats: Male Sprague-Dawley rats are used to assess the effects of APD334 on blood lymphopenia. Male rats are administered an oral dose of APD334 formulated in 0.5% methylcellulose (MC) in water at a rate of 0 mg/kg (vehicle only), 0.03 mg/kg (mice only), 0.1, 0.3, or 1 mg/kg. Samples of rat blood are taken 0, 1, 3, 5, 8, 16, 24, 32, 48, and 72 hours after the dose[1].
Mice: Male BALB/c mice are used to study the effects of APD334 on blood lymphopenia. In summary, APD334 is administered orally to male mice at doses of 0 (vehicle only), 0.03 (mice only), 0.1, 0.3, or 1 mg/kg when formulated in 0.5% methylcellulose (MC) in water. At 0, 1, 3, 5, 8, 16, 24, and 32 hours after administration, mouse blood samples are collected[1].

Absorption, Distribution and Excretion
Etrasimod mean (SD) steady-state maximum plasma concentration (Cmax) was 113 (27.5) ng/mL and the area under the time concentration curve at the dosing interval (AUCtau) was 2162 (488) ng*h/mL at the recommended dosage. Etrasimod Cmax and AUC are approximately dose-proportional from 0.7 mg to 2 mg (0.35 times up to the recommended dosage). Etrasimod steady state is reached within 7 days with an accumulation of approximately 2- to 3-fold compared to the first dose. The median (range) time to reach etrasimod Cmax (Tmax) is approximately 4 hours (range 2 to 8 hours) after oral administration. No clinically significant differences in the pharmacokinetics of etrasimod were observed following administration of etrasimod with a high-fat meal (800 to 1000 calories).
Approximately 82% of the total radioactive etrasimod dose was recovered in the feces and 5% in the urine within 336 hours. Approximately 11% of the administered radioactive dose was excreted as unchanged etrasimod in feces and none was excreted unchanged in urine.
The mean apparent volume of distribution of etrasimod is 66 (24) L.
The apparent steady-state oral clearance of etrasimod is approximately 1 L/h after oral administration.

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Metabolism / Metabolites
Etrasimod is metabolized by oxidation and dehydrogenation mediated primarily by CYP2C8, CYP2C9, and CYP3A4, with a minor contribution by CYP2C19 and CYP2J2. Etrasimod also undergoes conjugation primarily mediated by UGTs, with a minor contribution by sulfotransferases. Unchanged etrasimod is the main circulating component in plasma.

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Biological Half-Life
The mean plasma elimination half-life (t_1/2) of etrasimod is approximately 30 hours.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the use of etrasimod during breastfeeding. Because etrasimod is 98% bound to plasma proteins, the amount in milk is likely to be low. If the mother requires etrasimod, it is not a reason to discontinue breastfeeding. Until more data become available, etrasimod should be used with caution during breastfeeding, especially while nursing a newborn or preterm infant.
◉ 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.

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Protein Binding
Etrasimod plasma protein binding is 97.9%.

[1]. Discovery of APD334: Design of a Clinical Stage Functional Antagonist of the Sphingosine-1-phosphate-1 Receptor. ACS Med Chem Lett. 2014 Nov 4;5(12):1313-7.

Etrasimod is an organic heterotricyclic compound that is 1,2,3,4-tetrahydrocyclopenta[b]indole substituted by carboxymethyl and [4-cyclopentyl-3-(trifluoromethyl)phenyl]methoxy groups at positions 3R and 7, respectively. It is a potent and functional antagonist of the sphingosine-1-phosphate-1 (S1P1) receptor (IC50 = 1.88 nM in CHO cells). It has a role as an antibacterial agent, a sphingosine-1-phosphate receptor 1 antagonist, an immunosuppressive agent and an anti-inflammatory drug. It is a member of (trifluoromethyl)benzenes, a member of cyclopentanes, an aromatic ether, a monocarboxylic acid and an organic heterotricyclic compound. It is a conjugate acid of an etrasimod(1-).
Etrasimod is a synthetic next-generation selective Sphingosine 1-phosphate (S1P) receptor modulator that targets the S1P1,4,5 with no detectable activity on S1P2 and S1P3 receptors. S1P receptors are membrane-derived lysophospholipid signaling molecules that are involved in the sequestration of circulating peripheral lymphocytes in lymph nodes. Therefore, S1P receptor modulators like etrasimod were investigated in treating immune-mediated diseases like ulcerative colitis where a high level of inflammatory T cells is present in the gastrointestinal tract, thus causing diffuse mucosal inflammation. In fact, it has been observed that antigen-activated T cells within peripheral lymphoid organs can transiently downregulate S1P receptor levels to facilitate immune cells trafficking into the intestinal mucosa. Etrasimod was approved on October 13, 2023, by the FDA under the brand name VELSIPITY for the treatment of adults with moderately to severely active ulcerative colitis. This approval was based on favorable results obtained from Pfizer’s Elevate UC Phase III registrational program, consisting of the Elevate UC 52 and Elevate UC 12 clinical trials, that investigates the efficacy of a 2-mg daily dose regimen of etrasimod, with a 32% and 26% remission rate observed in UC 52 and UC 12 trials respectively.

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Drug Indication
Etrasimod is indicated for the treatment of moderately to severely active ulcerative colitis (UC) in adults.

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Mechanism of Action
Etrasimod is a sphingosine 1-phosphate (S1P) receptor modulator that binds with high affinity to S1P receptors 1, 4, and 5 (S1P_1,4,5). Etrasimod has minimal activity on S1P₃ (25-fold lower than C_max at the recommended dose) and no activity on S1P₂. Etrasimod partially and reversibly blocks the capacity of lymphocytes to egress from lymphoid organs, reducing the number of lymphocytes in peripheral blood. The mechanism by which etrasimod exerts therapeutic effects in UC is unknown but may involve the reduction of lymphocyte migration into the intestines.

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Pharmacodynamics
Etrasimod causes a reduction in peripheral blood lymphocyte count. In UC-1 and UC-2, mean lymphocyte counts decreased to approximately 50% of baseline at 2 weeks (approximate mean blood lymphocyte counts 0.9 x 109/L) and the lower lymphocyte counts were maintained during treatment with etrasimod. Dose-response relationship analysis indicates there is a dose-dependent reduction in blood lymphocyte counts. After discontinuing etrasimod 2 mg once daily, the median time for peripheral blood lymphocytes to return to the normal range was 2.6 weeks, with approximately 90% of subjects in the normal range within 4.7 weeks. Etrasimod may result in a transient decrease in heart rate and AV conduction upon treatment initiation. In UC-1 and UC-2, the mean (SD) decrease in heart rate was 7.2 (8.98) bpm at 2 to 3 hours after the first dose of etrasimod on Day 1. At 2 times the maximum recommended dose, etrasimod does not cause clinically significant QTc interval prolongation. Reductions in absolute FEV1 were also observed in subjects treated with etrasimod.

C26H26F3NO3

457.49321

457.186

C, 68.26; H, 5.73; F, 12.46; N, 3.06; O, 10.49

1206123-37-6

1206123-37-6; 1206123-97-8 (arginine)

44623998

White to khaki solid powder

1.3±0.1 g/cm3

621.4±50.0 °C at 760 mmHg

329.6±30.1 °C

0.0±1.9 mmHg at 25°C

1.606

6.43

62.3

2

6

6

33

695

1

FC(F)(F)C1=CC(COC2=CC=C(NC3=C4CC[C@@H]3CC(O)=O)C4=C2)=CC=C1C5CCCC5

MVGWUTBTXDYMND-QGZVFWFLSA-N

InChI=1S/C26H26F3NO3/c27-26(28,29)22-11-15(5-8-19(22)16-3-1-2-4-16)14-33-18-7-10-23-21(13-18)20-9-6-17(12-24(31)32)25(20)30-23/h5,7-8,10-11,13,16-17,30H,1-4,6,9,12,14H2,(H,31,32)/t17-/m1/s1

2-[(3R)-7-[[4-cyclopentyl-3-(trifluoromethyl)phenyl]methoxy]-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetic acid

APD 334; APD334; APD-334; Etrasimod; Velsipity

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO: ≥ 28 mg/mL (~61.2 mM)

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。