Physicochemical Properties
| Molecular Formula | C28H24FN5O2 |
| Molecular Weight | 481.52 |
| CAS # | 2820426-92-2 |
| Appearance | Typically exists as solid at room temperature |
| 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
| Targets | IC50: 0.7 nM (BTK)[1] |
| ln Vitro | BTK inhibition is a successful strategy against cancers of the B cell[1]. With an IC50 value of 2.6 nM, BTK-IN-15 (compound 42) exhibits inhibitory activity against TMD8[1]. With 0.05% and 44% of the control, respectively, BTK-IN-15 (1 μM; 1 h) shows notable selectivity to BTK over EGFR kinase[1]. BTK-IN-15 (0-1 mM; 72 h) has an IC50 value of 1.7 nM and demonstrates strong anti-proliferative action against the human mantle cell lymphoma cell line (REC-1)[1]. BTK-IN-15 (0-1 mM; 2 h) has an IC50 value of 1.49 nM[1], which inhibits BTK auto-phosphorylation. BTK-IN-15 (0–100 nM; 48 h) stops the cell cycle at the G1 phase, and in TMD8 (0–1 mM; 72 h), it causes apoptosis[1]. Low cardiotoxicity is shown by BTK-IN-15's low hERG channel activity (IC50=4.38 μM)[1]. |
| ln Vivo | At a dosage of 50 mg/kg in mice, BTK-IN-15 (compound 42) (12.5-50 mg/kg; po; twice daily; 21 d) suppresses tumor growth (TGI = 104%)[1]. In mice, BTK-IN-15 (300, 400, and 500 mg/kg; po; twice daily; 14 d) exhibits no effect on body weight when compared to control[1]. It is biologically safe. Mice treated with BTK-IN-15 (10 mg/kg; po) exhibit a high oral bioavailability of 40.98%[1]. BTK-IN-15 pharmacokinetics in mice[1] Path Dosage (g/kg) Tmax in hours Cmax in ng/mL AUC(0-t) (h·ng/mL) AUC(0-∞) (h·ng/mL) T1/2 (hour) Vz = L/kg CL in terms of L/h/kg F (%) IV 2 0.03 2245.39 1471.35 718.33 0.67 2.79 2.87 40.98 po 10 0.39 1441.59 718.33 1472.06 0.59 |
| Cell Assay |
Western Blot Analysis[1] Cell Types: B-cell lymphoma (DLBCL) TMD8 cancer cells Tested Concentrations: 0, 0.15, 0.46, 1.37, 4.12, 12.35, 37.04, 111, 333, 1000 nM Incubation Duration: 2 hrs (hours) Experimental Results: Inhibited BTK auto-phosphorylation at the Tyr223 site with an IC50 value of 1.49 nM. Cell Cycle Analysis[1] Cell Types: B-cell lymphoma (DLBCL) TMD8 cancer cells Tested Concentrations: 0, 10, 100 nM Incubation Duration: 48 hrs (hours) Experimental Results: Arrested cell cycle progression at G1 phase in a dose-dependent manner, the percentage of cells in the G0/G1 phase increased from 33.0 to 63.0% with a dose range of 1-100 nM. Apoptosis Analysis[1] Cell Types: B-cell lymphoma (DLBCL) TMD8 cancer cells Tested Concentrations: 0, 10, 100, 1000 nM Incubation Duration: 72 hrs (hours) Experimental Results: Induced apoptosis of TMD8 cells in a weakly triggered concentration-dependent manner, with the apoptosis cell values of 19% (10 nM), 25.2% (100 nM), and 31.4% (1000 nM), respectively. |
| Animal Protocol |
Animal/Disease Models: Female CB-17 SCID nude mice with TMD8 xenograft model[1] Doses: 12.5 mg/kg, 25 mg/kg, and 50 mg /kg Route of Administration: Oral gavage; twice (two times) daily; 21 days Experimental Results: Inhibited tumor growth at a dosage of 50 mg/kg and decreased tumor volume after 21 days with a TGI of 104%. decreased the content of white blood cells, lymphocytes and monocytes, while demonstrated no effect on red blood cells and platelets. Animal/Disease Models: ICR mice (acclimation for 5 days, 18-20 g)[1] Doses: 300, 400, 500 mg/kg Route of Administration: Oral gavage; 14 days Experimental Results: Demonstrated no affect against body weight in mice compared with control. |
| References |
[1]. Design, synthesis, and biological evaluation of pyrrolopyrimidine derivatives as novel Bruton's tyrosine kinase (BTK) inhibitors. EUR J MED. 2022 Jul. 114611. |
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 | 2.0768 mL | 10.3838 mL | 20.7676 mL | |
| 5 mM | 0.4154 mL | 2.0768 mL | 4.1535 mL | |
| 10 mM | 0.2077 mL | 1.0384 mL | 2.0768 mL |