| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
RP-001 is a selective agonist of the Sphingosine 1-Phosphate Receptor 1 (S1P1). It activates S1P1 in vitro with an EC50 of 9 pM. It has little activity on S1P2, S1P3, and S1P4, and only moderate affinity for S1P5 [1].
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| ln Vitro |
RP-001 induces a dose-dependent lymphopenia of CD4+ T cells. Acute lymphopenia is induced by RP-001, although it quickly returns to untreated levels [1].
RP-001 activated S1P1 in vitro with an EC50 of 9 pM, while having little activity on S1P2, S1P3, and S1P4, and only moderate affinity for S1P5. No differences were observed between the racemate and the resolved S-enantiomer across all five S1P receptors, demonstrating no chiral preference [1]. RP-001 induced sustained signaling while in internalized vesicles, as reported for FTY720, in contrast to the natural ligand S1P. It also induced dose-dependent internalization and polyubiquitination of S1P1 in vitro similar to other S1P1 agonists [1]. |
| ln Vivo |
The EC50 of RP-001 is 0.03 mg/kg, and it causes dose-dependent fast lymphopenia[1].
In vivo, RP-001 caused dose-dependent rapid lymphopenia with an EC50 of 0.03 mg/kg. Maximal lymphocyte sequestration from blood occurred within 2 hours and returned to untreated levels by 8 hours following treatment with 0.3 mg/kg [1]. In Edg1eGFP/eGFP knock-in mice, treatment with RP-001 (0.1 mg/kg) resulted in significant decreases in S1P1-eGFP expression on lymphocytes in the lymph node within 2 hours. Lymphocytic S1P1-eGFP did not fully recover to untreated levels by 8 hours after treatment, despite restoration of normal lymphocyte recirculation [1]. On endothelial cells in the lung, RP-001 (0.1 mg/kg) caused a small loss of S1P1-eGFP within 2 hours, but substantial degradation was observed 8 hours after treatment, when no agonist was present in circulation [1]. Using two-photon microscopy, treatment with RP-001 at 0.1 mg/kg did not lead to internalization of S1P1-eGFP on endothelial cells within the lymph node within 80 minutes. However, ex vivo incubation of explanted lymph nodes with a concentration of RP-001 (250 nM, 10-fold higher than the Cmax at 0.1 mg/kg) induced rapid ligand-induced internalization of S1P1-eGFP on endothelial cells [1]. |
| Cell Assay |
S1P1 Activation Assay (CRE-β-lactamase): CHO-K1 cells expressing S1P1 and a CRE-β-lactamase reporter were used. Cells were pretreated with 1 μM of the indicated compounds for 1 hour, washed, and rested for 5 hours. Cells were then treated with 2 μM forskolin with or without 10 mM FTY720 as indicated. Cyclic AMP-induced beta-lactamase expression was detected by cleavage of the fluorogenic substrate CC4-AM [1].
S1P1 Internalization and Polyubiquitination: In vitro assays demonstrated that RP-001 induced dose-dependent internalization and polyubiquitination of S1P1, similar to other S1P1 agonists [1]. |
| Animal Protocol |
Lymphopenia Studies: Mice were treated with RP-001 via intraperitoneal injection. Doses used included 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, and 1 mg/kg. Blood samples were collected at various time points (2 hours, 8 hours) to measure lymphocyte counts. For recovery studies, mice were treated with 0.3 mg/kg, and blood was collected at 2, 4, 6, and 8 hours post-injection. Blood concentration of RP-001 was quantified by mass spectrometry [1].
Endothelial Cell Studies: Mice were treated with RP-001 (0.1 mg/kg) via intraperitoneal injection. Lymph nodes and lungs were harvested at 2 hours and 8 hours post-injection for flow cytometric analysis of S1P1-eGFP expression on blood and lymphatic endothelial cells [1]. Ex Vivo Lymph Node Incubation: Explanted lymph nodes were incubated in the presence of 250 nM RP-001 (a concentration 10-fold higher than the Cmax at 0.1 mg/kg) to observe ligand-induced internalization of S1P1-eGFP on endothelial cells [1]. Two-Photon Microscopy: Mice were treated with 0.1 mg/kg RP-001 via intraperitoneal injection. Lymph nodes were imaged intravitally using two-photon microscopy to observe subcellular localization of S1P1-eGFP over time (up to 80 minutes) [1]. |
| ADME/Pharmacokinetics |
RP-001 has a short half-life in vivo. Following treatment with 0.3 mg/kg, blood concentration of RP-001 was quantified by mass spectrometry. The lower limit of detection was 1 nM. The short half-life renders RP-001 unsuitable for clinical use but allows examination of changes in S1P1 expression during both induction and recovery of lymphopenia [1].
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| References | |
| Additional Infomation |
Background: S1P1 is a high-affinity GPCR for S1P, essential for vascular development and integrity, and modulates lymphocyte development and recirculation. FTY720 (fingolimod), a prodrug whose phosphorylated form is an S1P1 agonist, was the first oral treatment for relapsing-remitting multiple sclerosis. However, FTY720 has a long duration of action and is thought to strongly downmodulate S1P1 expression [1].
Mechanism of Action: RP-001 is an orthosteric S1P1 agonist, competitive with the selective S1P1 antagonist W146. It induces sustained signaling while in internalized vesicles, similar to FTY720, and induces dose-dependent internalization and polyubiquitination of S1P1 [1]. Derivation: RP-001 was derived from the allosteric S1P1 agonist CYM-5442 by adding S1P-like headgroup interactions, which enhanced potency by 100-fold and shifted ligand binding into an orthosteric binding mode [1]. Purpose: Due to its short duration of action, RP-001 was used as a tool compound to elucidate physiological and agonist-perturbed changes in S1P1 expression at a subcellular level in vivo, particularly to demonstrate differential downregulation of S1P1 on lymphocytes and endothelia following agonist treatment [1]. |
| Molecular Formula |
C??H??N?O?
|
|---|---|
| Molecular Weight |
468.932704687119
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| Exact Mass |
468.156
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| CAS # |
1306761-53-4
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| Related CAS # |
RP-001 hydrochloride;1781880-34-9
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| PubChem CID |
90488927
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| Appearance |
Typically exists as solid at room temperature
|
| LogP |
5.306
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
33
|
| Complexity |
695
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC(C)OC1=C(C=C(C=C1)C2=NC(=NO2)C3=C4CCC(C4=CC=C3)NCCC(=O)O)C#N.Cl
|
| InChi Key |
ZHPGSGYEFGEZAL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H24N4O4.ClH/c1-14(2)31-21-9-6-15(12-16(21)13-25)24-27-23(28-32-24)19-5-3-4-18-17(19)7-8-20(18)26-11-10-22(29)30;/h3-6,9,12,14,20,26H,7-8,10-11H2,1-2H3,(H,29,30);1H
|
| Chemical Name |
3-[[4-[5-(3-cyano-4-propan-2-yloxyphenyl)-1,2,4-oxadiazol-3-yl]-2,3-dihydro-1H-inden-1-yl]amino]propanoic acid;hydrochloride
|
| Synonyms |
RP001; RP-001; 3-((4-(5-(3-Cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)amino)propanoic acid; orb1708631; SCHEMBL1878411; RP 001
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1325 mL | 10.6626 mL | 21.3251 mL | |
| 5 mM | 0.4265 mL | 2.1325 mL | 4.2650 mL | |
| 10 mM | 0.2133 mL | 1.0663 mL | 2.1325 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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