S1P1 ( IC50 = 1.88 nM )

Etrasimod arginine/APD-334 has a relatively low systemic clearance (<4% of hepatic blood flow) and high Cmax across all species. A notable reduction in volume of distribution (Vss) is noted in dogs and monkeys when compared to rodents. Oral bioavailability ranges from 40 to 100%, and the terminal phase half-life in dogs can reach up to 29 hours, while in monkeys it only lasts 6 hours. The t1/2 values for siponimod, an additional S1P1 modulator presently undergoing human trials, have been revealed to be 6 and 19 hours, respectively, for rats and monkeys[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
At the recommended dose, the mean (standard deviation) steady-state maximum plasma concentration (Cmax) of ectomod is 113 (27.5) ng/mL, and the area under the time-concentration curve (AUCtau) over the dosing interval is 2162 (488) ngh/mL. The Cmax and AUC of ectomod are approximately dose-proportional in the range of 0.7 mg to 2 mg (up to 0.35 times the recommended dose). Steady-state plasma concentrations of ectomod are reached within 7 days, representing a cumulative effect of approximately 2 to 3 times compared to the first dose. The median time to reach Cmax (Tmax) after oral administration is approximately 4 hours (range 2 to 8 hours). No clinically significant differences in the pharmacokinetics of ectomod were observed when co-administered with a high-fat meal (800 to 1000 calories).
Within 336 hours, approximately 82% of the total radioactive ezetimidate dose was recovered in feces and 5% in urine. Approximately 11% of the administered radioactive dose was excreted unchanged in feces, and unchanged ezetimidate was not detected in urine.
The mean apparent volume of distribution of ezetimidate was 66 (24) L.
The apparent steady-state oral clearance of ezetimidate after oral administration was approximately 1 L/h.

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Metabolism/Metabolites
Ezetimidate is primarily metabolized via oxidation and dehydrogenation mediated by CYP2C8, CYP2C9, and CYP3A4, with less contribution from CYP2C19 and CYP2J2. Ezetimidate is also primarily metabolized via UGT-mediated binding reactions, with less contribution from sulfotransferases. Unaltered ezetimidate is the major circulating component in plasma.

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

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
There is currently no information regarding the use of ezetitial during lactation. Because ezetitial binds to plasma proteins at a rate of 98%, its concentration in breast milk may be very low. If a mother needs to take ezetitial, she should not discontinue breastfeeding. Until more data are available, ezetitial should be used with caution during lactation, especially when breastfeeding newborns or premature infants. ◉ Effects on Breastfed Infants
No published information found as of the revision date. ◉ Effects on Lactation and Breast Milk
No published information found as of the revision date.

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Protein Binding Rate
Eezetitial has a plasma protein binding rate of 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 heterocyclic tricyclic compound with the structure 1,2,3,4-tetrahydrocyclopentano[b]indole, substituted at the 3R and 7 positions with carboxymethyl and [4-cyclopentyl-3-(trifluoromethyl)phenyl]methoxy, respectively. It is a potent functional antagonist of sphingosine-1-phosphate receptor 1 (S1P1) (IC50 = 1.88 nM in CHO cells). It possesses various pharmacological activities, including antibacterial, sphingosine-1-phosphate receptor 1 antagonism, immunosuppression, and anti-inflammatory effects. Etrasimod belongs to the (trifluoromethyl)benzene class, cyclopentane class, aromatic ether class, monocarboxylic acid class, and organic heterocyclic tricyclic compound class. It is the conjugate acid of ezetimibe (1-). Etrasimod is a synthetic, next-generation selective sphingosine-1-phosphate (S1P) receptor modulator that targets S1P1, 4, and 5 receptors, with no significant activity against S1P2 and S1P3 receptors. S1P receptors are membrane-derived lysophospholipid signaling molecules involved in the retention of circulating peripheral lymphocytes in lymph nodes. Therefore, S1P receptor modulators like ectomod are used to treat immune-mediated diseases such as ulcerative colitis, in which a large number of inflammatory T cells exist in the gastrointestinal tract, leading to diffuse mucosal inflammation. In fact, antigen-activated T cells in peripheral lymphoid organs have been observed to transiently downregulate S1P receptor levels to promote the migration of immune cells to the intestinal mucosa. On October 13, 2023, the FDA approved Etrasimod under the brand name VELSIPITY for the treatment of adult patients with moderate to severe active ulcerative colitis. This approval is based on positive results from Pfizer's Phase III Elevate UC registration study, which included two clinical trials, Elevate UC 52 and Elevate UC 12, to investigate the efficacy of a daily dose of 2 mg ectomod. Remission rates of 32% and 26% were observed in the UC 52 and UC 12 trials, respectively. Indications: Eectomod is indicated for the treatment of moderate to severe active ulcerative colitis (UC) in adults. Mechanism of Action: Eectomod 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 very low activity against S1P3 (25-fold lower than Cmax at the recommended dose) and no activity against S1P2. Etrasimod partially and reversibly blocks the migration of lymphocytes from lymphoid organs, thereby reducing the number of peripheral blood lymphocytes. The mechanism by which ectremod exerts its therapeutic effect in ulcerative colitis (UC) is unclear, but it may be related to reducing lymphocyte migration to the intestine. Pharmacodynamics: Etrasimod reduces peripheral blood lymphocyte counts. In UC-1 and UC-2 models, the mean lymphocyte count decreased to approximately 50% of baseline at 2 weeks (mean blood lymphocyte count approximately 0.9 x 10⁹/L) and remained at a low lymphocyte count level during ectremod treatment. Dose-response analysis showed a dose-dependent decrease in blood lymphocyte count. Following discontinuation of once-daily 2 mg ectomod, the median time for peripheral blood lymphocytes to return to normal was 2.6 weeks, with approximately 90% of subjects returning to normal within 4.7 weeks. Ectomod treatment may cause a transient decrease in heart rate and atrioventricular conduction at the beginning of treatment. In the UC-1 and UC-2 studies, a mean (standard deviation) decrease in heart rate of 7.2 (8.98) bpm was observed 2 to 3 hours after the first dose of ectomod on day 1. Even administration of twice the maximum recommended dose of ectomod did not cause clinically significant QTc interval prolongation. A decrease in absolute FEV1 was also observed in subjects treated with ectomod.

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

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; etrasimodum; 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)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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