D-I03 (DI03) is a novel and potent RAD52 inhibitor (Kd = 25.8 µM) with anticancer activity. RAD52 is a member of the homologous recombination (HR) pathway that is important for maintenance of genome integrity. While RAD52 mutations alone do not significantly affect a mammal's phenotype, they can be fatal when combined with mutations in other genes, such as BRCA1, BRCA2, PALB2, and RAD51C, that result in ovarian and breast cancer that is inherited. As such, RAD52 might be a crucial target for cancer treatment. D-I03 inhibits the growth of BRCA1- and BRCA2-deficient cells and prevents SSA and D-loop formation that are dependent on RAD52. also prevents the development of RAD52 foci brought on by damage, but has no effect on RAD51 foci brought on by cisplatin.
Physicochemical Properties
| Molecular Formula | C23H36N6S |
| Molecular Weight | 428.6371 |
| Exact Mass | 428.27 |
| Elemental Analysis | C, 64.45; H, 8.47; N, 19.61; S, 7.48 |
| CAS # | 688342-78-1 |
| PubChem CID | 24761372 |
| Appearance | Off-white to light yellow solid powder |
| LogP | 3.6 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 30 |
| Complexity | 520 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | UXDGHRWOHOPKIL-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C23H36N6S/c1-5-27(6-2)11-10-24-23(30)25-19-8-9-21-20(17-19)18(4)16-22(26-21)29-14-12-28(7-3)13-15-29/h8-9,16-17H,5-7,10-15H2,1-4H3,(H2,24,25,30) |
| Chemical Name | 1-[2-(diethylamino)ethyl]-3-[2-(4-ethylpiperazin-1-yl)-4-methylquinolin-6-yl]thiourea |
| Synonyms | DI03; D I03; D-I03 |
| 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 | RAD52 ( Ki = 25.8 µM ) |
| ln Vitro |
D-I03 (0-10 μM; on days 1 and 3; Capan-1 and UWB1.289 cells) treatment preferentially inhibited the growth of UWB1.289 and Capan-1 cells in a concentration-dependent manner[1]. D-I03 inhibits the formation of RAD52 foci in BCR-ABL1-positive, BRCA1-deficient 32Dcl3 murine hematopoietic cell line, which expresses GFP-RAD52, in response to cisplatin. The percentage of cells with RAD52 foci drops from 38.7% to 171% in the presence of D-I03 (2.5 μM); concurrently, the percentage of cells treated with ciprofloxacin without foci rises from 48.4% to 71.9%. ? D-I03 has no effect on RAD51 foci that are brought on by cisplatin. D-I03 exhibits low genotoxicity as evidenced by its inability to induce either RAD51 or RAD52 foci in BRCA1-deficient cells when used alone [1]. |
| ln Vivo | D-I03 (50 mg/kg/day; intraperitoneal injection; daily; for 7 days; nu/nu mice) treatment slows the growth of MDA-MB-436 tumors that lack BRCA1. Talazoparib puls D-I03 exhibits negligible toxicity against normal tissues and organs and has no effect on the growth of tumors that are BRCA1-proficient[3]. The results of pharmacokinetic and toxicity studies show that D-I03 has a maximal concentration in peripheral blood of >1 μM at a maximal tolerated dose of ≥50 mg/kg and t1/2 of 23.4 hours[1]. |
| Cell Assay |
Cell Line: Capan-1 (BRCA2−) and UWB1.289 (BRCA1+) cells Concentration: 0 μM, 2.5 μM, 5 μM, or 10 μM Incubation Time: On days 1 and 3 Result: Preferentially suppressed the growth of Capan-1 and UWB1.289 cells. |
| Animal Protocol |
Nu/nu mice injected with BRCA1-deficient MDA-MB-436 cells 50 mg/kg/day Intraperitoneal injection; daily; for 7 days |
| References |
[1]. Targeting BRCA1- and BRCA2-deficient cells with RAD52 small molecule inhibitors. Nucleic Acids Res. 2016 May 19;44(9):4189-99. [2]. Small-Molecule Inhibitors Targeting DNA Repair and DNA Repair Deficiency in Research and Cancer Therapy. Cell Chem Biol. 2017 Sep 21;24(9):1101-1119. [3]. Simultaneous Targeting of PARP1 and RAD52 Triggers Dual Synthetic Lethality in BRCA-Deficient Tumor Cells. Cell Rep. 2018 Jun 12;23(11):3127-3136. |
Solubility Data
| Solubility (In Vitro) |
DMSO: 50~86 mg/mL (116.7~200.6 mM) Ethanol: ~10 mg/mL |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.83 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.83 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.83 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3330 mL | 11.6648 mL | 23.3296 mL | |
| 5 mM | 0.4666 mL | 2.3330 mL | 4.6659 mL | |
| 10 mM | 0.2333 mL | 1.1665 mL | 2.3330 mL |