| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 50mg | |||
| Other Sizes |
| Targets |
Emzadirib (RAD51-IN-2) targets RAD51 recombinase (IC50 = 0.3 μM for inhibiting RAD51-mediated DNA strand exchange activity) [1]
|
|---|---|
| ln Vitro |
In activation-induced cytidine deaminase (AID)-positive cells, emzadirib displays an EC50 of less than 1 μM [1].
- RAD51 enzyme inhibitory activity: Emzadirib (RAD51-IN-2) potently inhibited RAD51-mediated DNA strand exchange activity with an IC50 of 0.3 μM. It blocked the formation of RAD51 nucleoprotein filaments on single-stranded DNA (ssDNA), a key step in homologous recombination (HR) repair [1] - Antiproliferative activity: The compound inhibited the proliferation of various cancer cell lines with defects in DNA repair pathways. It showed potent activity against BRCA1/2-deficient cancer cells, including MDA-MB-436 (IC50 = 1.2 μM) and Capan-1 (IC50 = 1.5 μM), and moderate activity against BRCA-proficient cells (MCF-7, IC50 = 8.7 μM) [1] - Sensitization to DNA-damaging agents: Emzadirib (RAD51-IN-2) enhanced the cytotoxicity of cisplatin and olaparib in BRCA-deficient cancer cells. When combined with cisplatin (1 μM), it reduced the IC50 of cisplatin in MDA-MB-436 cells from 4.5 μM to 1.8 μM. Combination with olaparib (0.5 μM) increased apoptosis by 2.6-fold compared to olaparib alone [1] - Inhibition of homologous recombination repair: In DR-GFP reporter assay, Emzadirib (RAD51-IN-2) (2 μM) reduced HR repair efficiency by 68% in U2OS cells. It also decreased the formation of RAD51 foci in γ-irradiated HeLa cells (inhibition rate = 72% at 5 μM) [1] |
| ln Vivo |
- Antitumor efficacy in BRCA-deficient xenografts: In nude mice bearing MDA-MB-436 (BRCA1-deficient) xenografts, intraperitoneal administration of Emzadirib (RAD51-IN-2) (20 mg/kg, twice weekly for 3 weeks) resulted in a tumor growth inhibition rate of 65%. Combination with cisplatin (5 mg/kg, once weekly) further increased the inhibition rate to 83% [1]
- Tolerability in animals: No significant body weight loss (≤ 7%) or obvious toxicological signs were observed in treated mice at the effective dose [1] |
| Enzyme Assay |
- RAD51 DNA strand exchange assay: Recombinant human RAD51 protein was incubated with ssDNA and Emzadirib (RAD51-IN-2) at gradient concentrations (0.05-5 μM) in reaction buffer (pH 7.5) for 30 minutes. Double-stranded DNA (dsDNA) substrate was then added, and the mixture was incubated at 37°C for 1 hour. The reaction products were separated by agarose gel electrophoresis, and the intensity of the strand exchange product band was quantified to calculate IC50 [1]
- RAD51 nucleoprotein filament formation assay: Fluorescently labeled ssDNA was mixed with RAD51 and Emzadirib (RAD51-IN-2) (0.1-10 μM) in binding buffer. Fluorescence anisotropy was measured to evaluate the effect of the compound on RAD51-ssDNA complex formation [1] |
| Cell Assay |
- Cell viability assay: Cancer cells (MDA-MB-436, Capan-1, MCF-7) were seeded into 96-well plates at 5×10³ cells/well, treated with Emzadirib (RAD51-IN-2) (0.1-20 μM) for 72 hours. Cell viability was measured by tetrazolium salt-based assay, and IC50 values were calculated [1]
- Combination cytotoxicity assay: MDA-MB-436 cells were pre-treated with Emzadirib (RAD51-IN-2) (0.5-5 μM) for 2 hours, then treated with cisplatin (0.5-10 μM) or olaparib (0.1-5 μM) for 72 hours. Cell viability was measured, and combination index (CI) was calculated to assess synergism [1] - HR repair efficiency assay: U2OS cells stably expressing DR-GFP reporter were transfected with I-SceI expression plasmid, then treated with Emzadirib (RAD51-IN-2) (0.5-5 μM) for 48 hours. GFP-positive cells were quantified by flow cytometry to determine HR repair efficiency [1] - RAD51 foci formation assay: HeLa cells were irradiated with 4 Gy γ-rays, then treated with Emzadirib (RAD51-IN-2) (1-10 μM) for 8 hours. Cells were fixed, stained with RAD51 antibody, and foci were counted under fluorescence microscopy [1] - Clonogenic assay: MDA-MB-436 cells were treated with Emzadirib (RAD51-IN-2) (0.5-2 μM) for 24 hours, seeded into 6-well plates at 500 cells/well, and incubated for 14 days. Colonies were stained and counted, with inhibition rate calculated relative to control [1] |
| Animal Protocol |
- BRCA-deficient xenograft model: Female nude mice (6-8 weeks old) were subcutaneously injected with MDA-MB-436 cells (5×10⁶ cells/mouse). When tumors reached ~100 mm³, mice were randomly divided into four groups: vehicle control, Emzadirib (RAD51-IN-2) alone (20 mg/kg, ip, twice weekly), cisplatin alone (5 mg/kg, ip, once weekly), and combination group (n=6 per group) [1]
- Drug formulation and administration: Emzadirib (RAD51-IN-2) was dissolved in a mixture of DMSO, PEG400, and sterile water (volume ratio 1:3:6) to prepare the administration solution. Treatment lasted for 3 weeks, with tumor volume measured every 3 days and body weight recorded weekly [1] - Tumor analysis: At the end of treatment, mice were sacrificed, tumors were excised and weighed, and tumor tissues were collected for immunohistochemical analysis of RAD51 foci formation [1] |
| ADME/Pharmacokinetics |
Plasma protein binding rate: The plasma protein binding rate of Emzadirib (RAD51-IN-2) in human plasma was 88.5 ± 1.8% as determined by equilibrium dialysis [1] - In vitro metabolic stability: The compound showed moderate metabolic stability in human liver microsomes with a half-life (t1/2) of 3.2 hours and a metabolic clearance rate of 0.52 mL/min/mg protein [1] - In vivo pharmacokinetics in mice: After a single intraperitoneal injection of 20 mg/kg, the Cmax was 9.6 μM, the AUC₀₋₂₄h was 42.3 μM·h, the elimination half-life (t1/2) was 4.1 hours, and the volume of distribution (Vd) was 1.8 L/kg [1]
|
| Toxicity/Toxicokinetics |
Acute toxicity: After a single intraperitoneal injection of Emzadirib (RAD51-IN-2) in mice, no death or obvious toxic symptoms were observed within 14 days at the maximum tolerated dose (MTD) >100 mg/kg [1]. Subacute toxicity: After treatment with Emzadirib (RAD51-IN-2) (20 mg/kg, intraperitoneal injection, twice a week for 3 weeks), no significant changes were observed in blood routine parameters or liver and kidney function indicators (ALT, AST, creatinine, blood urea nitrogen). Histopathological examination of major organs showed no abnormal lesions [1].
|
| References | |
| Additional Infomation |
Emzadirib is an oral bioavailability inhibitor of the DNA damage repair protein RAD51, with potential antitumor and sensitizing effects. After oral administration, emzadirib can target, bind to and inhibit the activity of RAD51. This can prevent RAD51-mediated DNA damage repair in susceptible tumor cells and induce tumor cell apoptosis. RAD51 is a core protein involved in DNA double-strand break (DSB) homologous repair (HR) and is overexpressed in a variety of tumor cell types.
- Chemical classification: Emzadirib (RAD51-IN-2) is a small molecule RAD51 inhibitor belonging to the quinazoline derivative class of compounds [1] - Mechanism of action: This compound binds to the ATP-binding pocket of RAD51, inhibiting its ATPase activity and subsequent nucleoprotein filament formation, thereby blocking homologous recombination DNA repair. This selectively sensitizes DNA-repair-deficient cancer cells to apoptosis and enhances the efficacy of DNA-damaging chemotherapy drugs [1] - Therapeutic potential: It is a promising targeted drug for the treatment of cancers with DNA-repair deficiencies (e.g., breast cancer, ovarian cancer, and pancreatic cancer with BRCA1/2 mutations), especially when used in combination with DNA-damaging drugs or PARP inhibitors [1]. |
| Molecular Formula |
C27H40N4O6S2
|
|---|---|
| Molecular Weight |
580.76
|
| Exact Mass |
580.238
|
| CAS # |
2301085-04-9
|
| PubChem CID |
137542570
|
| Appearance |
White to off-white solid powder
|
| LogP |
4.8
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
11
|
| Heavy Atom Count |
39
|
| Complexity |
926
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1(C2SC(=NC=2)[C@@H]2CC[C@@H](NC(=O)OC(C)C)CC2)=C(S(=O)(=O)NC(C)(C)C)C=C(C=C1)NC(=O)OC(C)C
|
| InChi Key |
OVXFEICGTUUFPE-WGSAOQKQSA-N
|
| InChi Code |
InChI=1S/C27H40N4O6S2/c1-16(2)36-25(32)29-19-10-8-18(9-11-19)24-28-15-22(38-24)21-13-12-20(30-26(33)37-17(3)4)14-23(21)39(34,35)31-27(5,6)7/h12-19,31H,8-11H2,1-7H3,(H,29,32)(H,30,33)/t18-,19-
|
| Chemical Name |
isopropyl ((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-((isopropoxycarbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate
|
| Synonyms |
RAD51 IN-2RAD51-IN-2 RAD51 IN 2 RAD51-IN 2
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~172.19 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.30 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 2: ≥ 2.5 mg/mL (4.30 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 | 1.7219 mL | 8.6094 mL | 17.2188 mL | |
| 5 mM | 0.3444 mL | 1.7219 mL | 3.4438 mL | |
| 10 mM | 0.1722 mL | 0.8609 mL | 1.7219 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
