Physicochemical Properties
| Molecular Formula | C25H20CLFN8O |
| Molecular Weight | 502.93070602417 |
| Exact Mass | 502.143 |
| CAS # | 2421122-61-2 |
| PubChem CID | 162670784 |
| Appearance | Typically exists as solid at room temperature |
| LogP | 4.1 |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 36 |
| Complexity | 743 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | BWFYYTOGYBJCQR-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C25H20ClFN8O/c26-17-11-16(9-10-18(17)27)31-22-21-23(34-25(29)33-22)35(13-30-21)12-14-5-7-15(8-6-14)24(36)32-20-4-2-1-3-19(20)28/h1-11,13H,12,28H2,(H,32,36)(H3,29,31,33,34) |
| Chemical Name | 4-[[2-amino-6-(3-chloro-4-fluoroanilino)purin-9-yl]methyl]-N-(2-aminophenyl)benzamide |
| 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 | Triple-negative breast cancer cells MDA-MB-231 (IC50 = 1.48 μM), MDA-MB-468 (IC50 = 0.65 μM), and liver cancer cell HepG2 (IC50 = 2.44 μM) are all inhibited by HDAC-IN-45 (Compound 14). HDAC-IN-45 overcomes HDACi resistance and is equally toxic to drug-resistant gastric cell lines (YCC3/7) and HDAC-sensitive cell lines (YCC11). HDAC-IN-45 exhibits high toxicity (IC50 = 0.33 μM) towards K-562, KG-1, and THP-1 leukemia cell lines. Compound 14, HDAC-IN-45, exhibits a noteworthy inhibitory impact on the HDAC1, 2, and 3 isomers, as evidenced by its respective IC50 values of 0.108, 0.585, and 0.563 μM. Compound 14—HDAC-IN-45—has the ability to raise both the expression of p21 and the degree of acetylation of histone H3. In MDA-MB-231 cells, HDAC-IN-45 (compound 14) caused dose-dependent upregulation of ac-H3K9, resulting in cell cycle arrest in the G1 phase. In xenograft mouse models, HDAC-IN-45 (compound 14) demonstrates strong anti-tumor efficacy [1]. |
| ln Vivo | Compound 14 (HDAC-IN-45) administered intraperitoneally (i.p.) at doses of 25 mg/kg or 50 mg/kg on a daily basis exhibited strong antitumor efficaciousness in a human MDA-MB-231 breast cancer model [1]. |
| Cell Assay |
Cell proliferation experiment [1] Cell Types: triple-negative breast cancer cells; liver cancer cells; YCC11 and YCC3/7 Tested Concentrations: serial concentration Incubation Duration: 72 hrs (hours). Experimental Results: inhibited the cell growth activity of HepG2 and triple-negative breast cancer cells. Cytotoxicity assay[1] Cell Types: Three leukemia cell lines (K-562, KG-1 and THP-1); YCC3/7 and YCC11 cell lines Tested Concentrations: A range of concentrations Incubation Duration: 72 hrs (hours) Experimental Results: demonstrated to be effective Anti-cancer effect, showing high sensitivity and strong toxicity in leukemia cell lines, with IC50 value below micromolar. Western Blot Analysis[1] Cell Types: MDA-MB-231 Cell Tested Concentrations: 2 µM Incubation Duration: 24 hrs (hours) Experimental Results: Increased histone H3 acetylation levels and p21 expression. Cell cycle analysis[1] Cell Types: MDA-MB-231cells Tested Concentrations: 4 µM Incubation Duration: 24 hrs (hours) Experimental Results: G1 phase cell cycle arrest and apoptosis. |
| Animal Protocol |
Animal/Disease Models: Human MDA-MB-231 breast cancer xenograft mouse model [1] Doses: 25 mg/kg or 50mg/kg Route of Administration: 25 mg/kg or 50mg/kg; ip; daily. Experimental Results: Demonstrated potent anti-tumor efficacy. |
| References |
[1]. Purine/purine isoster based scaffolds as new derivatives of benzamide class of HDAC inhibitors. Eur J Med Chem. 2020 Jun 15;196:112291. |
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 | 1.9883 mL | 9.9417 mL | 19.8835 mL | |
| 5 mM | 0.3977 mL | 1.9883 mL | 3.9767 mL | |
| 10 mM | 0.1988 mL | 0.9942 mL | 1.9883 mL |