Physicochemical Properties
| Molecular Formula | C23H21F3N4O3 |
| Molecular Weight | 458.433055639267 |
| Exact Mass | 458.156 |
| CAS # | 2287259-07-6 |
| PubChem CID | 146014480 |
| Appearance | White to off-white solid powder |
| LogP | 4.2 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 7 |
| Heavy Atom Count | 33 |
| Complexity | 702 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | FC(C1C=CC(=CC=1)CN1C(=C(/C=C/C)N=N1)C(N[C@@H](C)C1C=CC(C(=O)O)=CC=1)=O)(F)F |
| InChi Key | ZTWUZRMXAVDXJV-XGACYXMMSA-N |
| InChi Code | InChI=1S/C23H21F3N4O3/c1-3-4-19-20(21(31)27-14(2)16-7-9-17(10-8-16)22(32)33)30(29-28-19)13-15-5-11-18(12-6-15)23(24,25)26/h3-12,14H,13H2,1-2H3,(H,27,31)(H,32,33)/b4-3+/t14-/m0/s1 |
| Chemical Name | 4-[(1S)-1-[[5-[(E)-prop-1-enyl]-3-[[4-(trifluoromethyl)phenyl]methyl]triazole-4-carbonyl]amino]ethyl]benzoic acid |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| ln Vitro | In CHO-Gα16 cells overexpressing human EP4 receptor, EP4 receptor antagonist 1 (compound 59) antagonized human EP4 with an IC50 of 6.1 ± 0.2 nM in the calcium flux assay. With an IC50 of 16.2±1.7 nM, EP4 receptor antagonist 1 antagonized human EP4 in the calcium flux experiment in CHO-Gα16 cells overexpressing the mouse EP4 receptor [1]. With an IC50 of 18.7±0.6 nM, EP4 receptor antagonist 1 dose-dependently prevents PGE2-stimulated cAMP buildup in HEK293-EP4 cells. With an IC50 of 5.2±0.4 nM, EP4 receptor antagonist 1 dose-dependently suppresses the expression of the CRE reporter gene in HEK293 cells. With an IC50 of 0.4±0.1nM, EP4 receptor antagonist 1 dose-dependently blocks PGE2-stimulated β-arrestin recruitment in HEK293-EP4 cells [1]. In a concentration-dependent manner, EP4 receptor antagonist 1 (1 nM-10 μM) reverses PGE2-induced ERK phosphorylation [1]. |
| ln Vivo | In BALB/c female mice, the oral administration of EP4 receptor antagonist 1 (16, 50, and 150 mg/kg; once daily for two weeks) greatly prevented the formation of tumors. Not a single mouse group showed any discernible weight loss. Mice respond well to EP4 receptor antagonist 1 at the tested doses [1]. In mice, EP4 receptor antagonist 1 (1 mg/kg; intraperitoneal) had a good half-life (t1/2) of 4.1 hours and moderate clearance (CL=1.7 L/h/kg). In mice, EP4 receptor antagonist 1 (5 mg/kg; oral) exhibits high bioavailability (F=48.0%) and a 4.7-hour half-life (t1/2). |
| Cell Assay |
Western Blot Analysis[1] Cell Types: CHO-EP4 Cell Tested Concentrations: 1 nM, 100 nM, 10 μM Incubation Duration: 20 minutes pretreatment, followed by 30 nM PGE2 simulation for 10 minutes. Experimental Results: PGE2-induced ERK phosphorylation was reversed in a concentration-dependent manner. |
| Animal Protocol |
Animal/Disease Models: BALB/c female mouse (6 weeks old) CT26 colon cancer model [1] Doses: 16, 50, 150 mg/kg Route of Administration: oral; one time/day for two weeks Experimental Results: Tumor growth inhibition (TGI ) is 24.6% at 16 mg/kg, 54.7% at 50 mg/kg, and 63.8% at 150 mg/kg. Animal/Disease Models: BALB/c female mice [1] Doses: 1 mg/kg and 5 mg/kg (pharmacokinetic/PK/PK analysis) Route of Administration: intravenous (iv) (iv)or po (po (oral gavage)) dose 1 mg/kg (5 mL/kg ) and 5 mg/kg (10 ml/kg), respectively. Experimental Results: At a dose of 1 mg/kg (iv), mice demonstrated moderate clearance (CL=1.7 L/h/kg) with a corresponding favorable half-life (t1/2) of 4.1 hrs (hrs (hours)). demonstrated good bioavailability in mice (F = 48.0%) with a corresponding favorable half-life (t1/2) of 4.7 hrs (hrs (hours)) at a dose of 5 mg/kg (oral). |
| References |
[1]. Discovery and Characterization of 1H-1,2,3-Triazole Derivatives as Novel Prostanoid EP4 Receptor Antagonists for Cancer Immunotherapy. J Med Chem. 2020 Jan 23;63(2):569-590. |
Solubility Data
| 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 |
| 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 (e.g. IP/IV/IM/SC) 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). 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) 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). 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 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1814 mL | 10.9068 mL | 21.8136 mL | |
| 5 mM | 0.4363 mL | 2.1814 mL | 4.3627 mL | |
| 10 mM | 0.2181 mL | 1.0907 mL | 2.1814 mL |