 Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C21H16F3N7O |
| Molecular Weight | 439.393253326416 |
| Exact Mass | 439.136 |
| CAS # | 1900754-56-4 |
| PubChem CID | 121273514 |
| Appearance | White to off-white solid powder |
| LogP | 1.7 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 9 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 32 |
| Complexity | 637 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | C1N(CC(NC2=NC=C(C3=NC=CN=C3)C=C2)=O)C(C)=C(C2C=CN=C(C(F)(F)F)C=2)N=1 |
| InChi Key | QMLOYDPILBUVBV-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C21H16F3N7O/c1-13-20(14-4-5-27-17(8-14)21(22,23)24)29-12-31(13)11-19(32)30-18-3-2-15(9-28-18)16-10-25-6-7-26-16/h2-10,12H,11H2,1H3,(H,28,30,32) |
| Chemical Name | 2-[5-methyl-4-[2-(trifluoromethyl)pyridin-4-yl]imidazol-1-yl]-N-(5-pyrazin-2-ylpyridin-2-yl)acetamide |
| 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 | Zamaporvint (300 nM, 48 h) treatment of L-wnt3a cells decreased, with an IC50 of 64 pM, the conditioned medium's ability to activate the β-catenin-responsive luciferase reporter gene. This effect was concentration-dependent. Luciferase activity was restored upon the addition of recombinant Wnt3a, suggesting that downstream Wnt signaling was unaffected [1]. The impact of Zamaporvint (100 nM, 24 h) on proliferation is indicative of c-Myc mRNA downregulation that is concentration-dependent. has been found to have decreased immunosuppression at the same dose as after administration The auxiliary role of sex [1]. It also decreases the percentage of cells in S phase and strongly inhibits the expression of the mitotic marker phospho-histone-H3 in cells with abnormalities upstream of Wnt pathway components, indicating cell cycle arrest. In plasma, zamaprovint (20 μM, 18 h) varies between 2.5% and 7.5% across species, while microsomal CLint values span from 3.9 to 31.6 μL/min.mg, with mice and dogs having the lowest expected clearance rates. has the greatest clearance rate, while human and rodent clearance rates are lower [1]. In MDR1-MDCKII cells, zamaporvint (10 μM, 2 h) exhibits some efflux and good intrinsic permeability, but not in Caco-2 cells [1]. |
| ln Vivo | In Wnt Inhibitory ligand-dependent SNU-1411, AsPC1, and HPAF II models, Zamaporvint (1.5 mg/kg or 5 mg/kg orally twice daily, or 5 mg/kg Zamaporvint orally once daily, for 28 days) reduces tumor growth and Wnt-responsive gene expression (including cMyc); tumor growth was unaffected in the Wnt ligand-independent HCT116 xenograft model[1]. In the B16F10 "cold" tumor model, zamaprovint (1.5 mg/kg, 5 mg/kg, once daily) inhibited immune evasion and decreased the number of Ki67-positive cells in the entire tumor area, with the impact being more noticeable in differentiated tumor areas[1]. Zamaporvint (1.5 or 5 mg/kg once daily) increases the percentage of regulatory T cells within CD8+/CT26 tumors, decreases B16F10 tumor-resident myeloid-derived suppressor cells, and collaborates with anti-programmed cell death protein-1 (PD-1, HY-P73361)[1]. Zamaporvint's pharmacokinetic characteristics in mice[1] |
| Cell Assay |
Western Blot Analysis[1] Cell Types: L-Wnt5a Tested Concentrations: 300 nM Incubation Duration: 48 h Experimental Results: Activated the β-catenin-responsive luciferase reporter gene in a concentration-dependent manner, with an IC50 of 64 pmol/L. Apoptosis Analysis[1] Cell Types: L-Wnt5a Tested Concentrations: 100 nM Incubation Duration: 24 h Experimental Results: Downregulated c-Myc mRNA and reduce the proportion of cells in S-phase, and strongly inhibited expression of the mitosis marker phospho-histone-H3 in cells with upstream aberrations in Wnt pathway components. |
| Animal Protocol |
Animal/Disease Models: SCID-Beige mice were dosed at Translational Drug Discovery with vehicle[1] Doses: 1.5 mg/kg or 5 mg/kg; 5 mg/kg Route of Administration: 1.5 mg/kg or 5 mg/kg orally twice (two times) daily, or 5 mg/kg RXC004 orally one time/day, for 28 days Experimental Results: decreased in tumor growth, and inhibition of Wnt-responsive gene expression including cMyc, was observed in the Wnt ligand– dependent SNU-1411, AsPC1, and HPAFII models. No effected tumor growth in the Wnt ligand–independent HCT116 xenograft mode. Animal/Disease Models: HPAF-II (5 × 106 cells; athymic nude mice), AsPC1 (3 × 106 cells; athymic nude mice), and SNU-1411 (1×107 cells; NOD-SCID mice) were implanted bilaterally, subcutaneously, whereas HCT116 (3 × 106 cells; athymic nude mice) were implanted in a single flank[1] Doses: Dosing was either 1.5 mg/kg twice (two times) daily RXC004 for 7–13 days then one time/day for the remainder of study (up to 29 days), or 28 days 1.5 mg/kg twice (two times) daily RXC004 for HCT116 Route of Administration: po Experimental Results: Demonstrated to inhibit tumor grow |
| References |
[1]. Phillips C, The Wnt Pathway Inhibitor RXC004 Blocks Tumor Growth and Reverses Immune Evasion in Wnt Ligand-dependent Cancer Models. Cancer Res Commun. 2022 Sep 2;2(9):914-928. |
| Additional Infomation | Zamaporvint is an orally available inhibitor of porcupine (PORCN), with potential antineoplastic activity. Upon oral administration, zamaporvint binds to and inhibits PORCN in the endoplasmic reticulum (ER), which blocks post-translational acylation of Wnt ligands and inhibits their secretion. This prevents the activation of Wnt ligands, interferes with Wnt-mediated signaling, and inhibits cell growth in Wnt-driven tumors. Porcupine, a membrane-bound O-acyltransferase (MBOAT), is required for the palmitoylation of Wnt ligands, and plays a key role in Wnt ligand secretion and activity. Wnt signaling is dysregulated in a variety of cancers. |
Solubility Data
| Solubility (In Vitro) | DMSO: 125 mg/mL (284.49 mM) |
| 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 | 2.2759 mL | 11.3794 mL | 22.7588 mL | |
| 5 mM | 0.4552 mL | 2.2759 mL | 4.5518 mL | |
| 10 mM | 0.2276 mL | 1.1379 mL | 2.2759 mL |