Physicochemical Properties
| Molecular Formula | C26H22CLN7O |
| Molecular Weight | 483.952183246613 |
| Exact Mass | 483.157 |
| CAS # | 2418559-01-8 |
| PubChem CID | 162674387 |
| Appearance | Typically exists as solid at room temperature |
| LogP | 5 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 6 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 35 |
| Complexity | 697 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | XHELPOUKGPDRRH-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C26H22ClN7O/c1-16-6-12-19(13-7-16)30-23-22-24(33-26(27)32-23)34(15-29-22)14-17-8-10-18(11-9-17)25(35)31-21-5-3-2-4-20(21)28/h2-13,15H,14,28H2,1H3,(H,31,35)(H,30,32,33) |
| Chemical Name | N-(2-aminophenyl)-4-[[2-chloro-6-(4-methylanilino)purin-9-yl]methyl]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 | HDAC1/2 and CDK2-IN-1 (compound 14d), with IC50 values of 1.59, 0.47, 0.86, 0.58, and 1.05 μM, respectively, demonstrate remarkable anti-proliferative actions against H460, A375, HepG2, HCT116, and HeLa cells [1]. The migration of H460 and A375 cells is strongly inhibited by HDAC1/2 and CDK2-IN-1 (0.5 μM, 48 h) [1]. In the G2/M phase, HDAC1/2 and CDK2-IN-1 (0–2 μM, 24 hours) can profoundly stop the cell cycle [1]. In a dose-dependent manner, HDAC1/2 and CDK2-IN-1 (0–2 μM, 48 hours) stimulate the death of cancer cells [1]. Cancer cells die as a result of CDK2 and HDAC activity inhibition by HDAC1/2 and CDK2-IN-1 (1 μM, 12 hours) [1]. In A375 cells, HDAC1/2 and CDK2-IN-1 (1 μM, 24 hours) significantly raise intracellular ROS levels, which ultimately results in cancer cell death [1]. |
| ln Vivo | Tumor growth was markedly inhibited by HDAC1/2 and CDK2-IN-1 (BALB/c nude mice, 0-100 mg/kg, IP, once daily for 21 days) [1]. The pharmacokinetic properties of HDAC1/2 and CDK2-IN-1 (compound 14d) (ICR mice; 4 mg/kg, IV; 20 mg/kg, IP) are ideal [1]. HDAC1/2 and CDK2-IN-1 pharmacokinetic parameters in male ICR mice [1]. T1/2 (h) 1.48 2.84 Tmax (h) 2 Cmax (ng/mL) 1360 AUC0-t (ng/mL*h) 2850 7240 MRT0-t (h) 0.563 4.54 CL (mL/(min/kg)) 23.3 F (%) 50.8; Dosage (mg/kg) 4 20 Intravenous |
| Cell Assay |
Cell cycle analysis Cell Types: A375, HCT116, H460 and Hela cells [1] Tested Concentrations: 0, 0.5, 1, 2 μM Incubation Duration: 24 hrs (hours) Experimental Results: Dramatically blocked cell cycle, induced loss of cells in G0/G1 phase, G2 /M phase cells increased, resulting in significant accumulation of G2/M phase cells at 0.5 μM (A375, the percentage increased from 13.70 to 57.03%; HCT116, increased from 27.46% to 76.99%; Hela, increased from 7.89% to 51.85) % ). Apoptosis analysis Cell Types: A375, HCT116, H460 and Hela cell lines [1] Tested Concentrations: 0, 0.5, 1, 2 μM Incubation Duration: 48 h Experimental Results: Promote cancer cell apoptosis in a dose-dependent manner, apoptosis rate When the concentration is 2 μM, they are 91.99% (A375), 89.60% (HCT116), 59.10% (H460) and 22.36% (Hela) respectively. Immunofluorescence Cell Types: A375 cells [1] Tested Concentrations: 1 μM Incubation Duration: 12 h Experimental Results: Dramatically inhibits CDK2 and increases the acetylation level of histone H3, inhibits CDK2 and HDAC activity, and leads to cancer ce |
| Animal Protocol |
Animal/Disease Models: Male ICR mouse (n = 9)[1] Doses: 4 mg/kg (IV), 20 mg/kg (IP) Mode of Route of Administration: IV, IP, once (pharmacokinetic/PK/PK analysis) Experimental Results: Demonstrated ideal pharmacokinetic/PK/PK properties. Animal/Disease Models: BALB/c nude mice (5-6 weeks, HCT116 xenograft model) [1] Doses: 0, 25, 50 and 100 mg/kg Route of Administration: IP, one time/day for 21 days Experimental Results: Significant Inhibit tumor growth. At doses of 25, 50 and 100 mg/kg, the tumor growth inhibition rates were 28%, 40% and 44% respectively. |
| References |
[1]. Yun F, Cheng C, Ullah S, Yuan Q. Design, synthesis and biological evaluation of novel histone deacetylase1/2 (HDAC1/2) and cyclin-dependent Kinase2 (CDK2) dual inhibitors against malignant cancer. Eur J Med Chem. 2020;198:112322. |
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 | 2.0663 mL | 10.3316 mL | 20.6633 mL | |
| 5 mM | 0.4133 mL | 2.0663 mL | 4.1327 mL | |
| 10 mM | 0.2066 mL | 1.0332 mL | 2.0663 mL |