Physicochemical Properties
| Molecular Formula | C29H30CLN5O5 |
| Molecular Weight | 564.03 |
| Exact Mass | 563.193 |
| CAS # | 2756851-99-5 |
| PubChem CID | 162641712 |
| Appearance | White to off-white solid powder |
| LogP | 3.7 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 40 |
| Complexity | 907 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O(C1C=C(OC)C=C2NC(C3C=CC(OCC(=O)NC4C=CC(N5CCN(C)CC5)=C(Cl)C=4)=CC=3)=NC(=O)C=12)C |
| InChi Key | UZWCUBDJHDWTFV-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C29H30ClN5O5/c1-34-10-12-35(13-11-34)24-9-6-19(14-22(24)30)31-26(36)17-40-20-7-4-18(5-8-20)28-32-23-15-21(38-2)16-25(39-3)27(23)29(37)33-28/h4-9,14-16H,10-13,17H2,1-3H3,(H,31,36)(H,32,33,37) |
| Chemical Name | N-[3-chloro-4-(4-methylpiperazin-1-yl)phenyl]-2-[4-(5,7-dimethoxy-4-oxo-3H-quinazolin-2-yl)phenoxy]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
| Targets | BRD4 180 nM (IC50) CK2 230 nM (IC50) |
| ln Vitro | Compound 44e, BRD4/CK2-IN-1 (0-25 μM; 24 hours) exhibits an anti-proliferation effect in MDA-MB-231 and MDA-MB-468 cells, with IC50s of 2.66 and 3.52 μM, respectively[1]. BRD4/CK2-IN-1 (0-10 μM; 24 hours) dose-dependently promotes apoptosis in MDA-MB-231 and MDA-MB-468 cells. Bcl-2 is dose-dependently downregulated by BRD4/CK2-IN-1 (0–10 μM; 24 hours), while Bax and cleaved caspase-3 are upregulated[1]. In MDA-MB-231 and MDA-MB, BRD4/CK2-IN-1 (0–10 μM; 24 hours) markedly downregulates the autophagy substrate p62 and upregulates beclin-1 and LC3II[1]. |
| ln Vivo | In TNBC xenograft models, BRD4/CK2-IN-1 (25 and 50 mg/kg; intragastric administration; daily for 19 days) inhibits tumor growth[1]. In the MDA-MB-231 and MDA-MB-468 xenograft models, BRD4/CK2-IN-1 (25 and 50 mg/kg; intragastric administration; daily for 19 days) exhibits mild toxicity as determined by body weight loss[1]. Pharmacokinetics (PK) profile of BRD4/CK2-IN-1: A preliminary assessment [1]. Parameter: iv (1 mg/kg) po (10 mg/kg) T1/2 (h) 4.21±0.57 5.14±0.71 Cmax (ng/mL) 237±11 206±6 AUC0-t (ng·h/ mL) 579±49 2079±130 AUC0-∞ (ng·h/mL) 588±36 2090±146 VZ (L/kg) 21.1±2.6 CL ((mL/min)/kg) 57.4±1.3 F (%) 32.5 |
| Cell Assay |
Cell Viability Assay[1] Cell Types: MDA-MB-231, MDA-MB-468 cells Tested Concentrations: 0, 0.19, 0.39, 0.78, 1.56, 3.13, 6.25, 12.5, 25 μM Incubation Duration: 24 hrs (hours) Experimental Results: demonstrated anti-proliferative rates in MDA-MB-231 and MDA-MB-468 cells (IC50 = 2.66 and 3.52 μM, respectively). Apoptosis Analysis[1] Cell Types: MDA-MB-231, MDA-MB-468 cells Tested Concentrations: 0, 2.5, 5, 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Dose-dependently induced apoptosis of MDA-MB-231 and MDA-MB-468 cells. Western Blot Analysis[1] Cell Types: MDA-MB-231, MDA-MB-468 cells Tested Concentrations: 0, 2.5, 5, 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Dose-dependently downregulated Bcl-2 but upregulated Bax and cleaved caspase-3. Cell Types: Tested Concentrations: Incubation Duration: Experimental Results: |
| Animal Protocol |
Animal/Disease Models: Female nude mice (BALB/c, 6-8 weeks, 20-22 g) bearing MDA-MB-231 cells[1] Doses: 25 and 50 mg/kg Route of Administration: intragastric (po)administration; daily for 19 days Experimental Results: Had the most pronounced tumor growth inhibition (TGI) (63.8%) in the MDA-MB-231 xenograft tumor model at 50 mg/kg. |
| References |
[1]. Discovery of Novel Dual-Target Inhibitor of Bromodomain-Containing Protein 4/Casein Kinase 2 Inducing Apoptosis and Autophagy-Associated Cell Death for Triple-Negative Breast Cancer Therapy. J Med Chem. 2021;64(24):18025-18053. |
Solubility Data
| Solubility (In Vitro) | DMSO : 20 mg/mL (35.46 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 | 1.7730 mL | 8.8648 mL | 17.7296 mL | |
| 5 mM | 0.3546 mL | 1.7730 mL | 3.5459 mL | |
| 10 mM | 0.1773 mL | 0.8865 mL | 1.7730 mL |