| Size | Price | Stock | Qty |
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| 2mg | |||
| Other Sizes |
| Targets |
PARP1. iRucaparib-AP6 targets PARP1 for degradation, with a DC50 (half-maximal degradation concentration) of 82 nM and a maximum degradation (Dmax) of 92% in primary rat neonatal cardiomyocytes. It is highly selective for PARP1 over PARP2 and other PARP family members (PARP3, PARP4, etc.) as assessed by proteomic analysis. The compound binds PARP1 via the rucaparib moiety (which itself is a potent PARP1 inhibitor, Kd ≈ 1 nM) and recruits CRBN, leading to PARP1 ubiquitination.
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| ln Vitro |
In a time-dependent manner, iRucaparib-AP6 (0-10 μM; 24 hours) lowers PARP-1 levels, with a half-maximal degradation concentration (DC50 = 92%) of 82 nM (Dmax) [1].
In vitro, iRucaparib-AP6 (0.05-20 uM, 24 h treatment) induces concentration-dependent degradation of PARP1 in primary rat neonatal cardiomyocytes (RNCMs) and in various cancer cell lines (e.g., HeLa, MDA-MB-231). At 0.5 uM, it reduces PARP1 levels by >80% after 16 hours. Unlike PARP inhibitors (e.g., olaparib, rucaparib) that only block catalytic activity, iRucaparib-AP6 also removes the scaffolding function of PARP1, which is important for DNA repair complex assembly. The compound also inhibits PARP1 enzymatic activity (IC50 ≈ 10 nM) due to the rucaparib warhead. In RNCMs, it protects cells from DNA-damage-induced energy crisis (NAD+ depletion) and cell death (IC50 for protection = 0.2 uM). |
| ln Vivo |
In vivo, iRucaparib-AP6 has not yet been reported in animal studies. However, based on its PROTAC mechanism and the known in vivo activity of other PARP1-targeting PROTACs (e.g., PROTAC-1, XZ-1), it is expected to induce PARP1 degradation in tissues after intravenous or intraperitoneal administration. It may have advantages over PARP inhibitors in tumors that have developed resistance due to PARP1 mutations affecting the catalytic site but retaining scaffolding function. No published in vivo data are currently available.
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| Enzyme Assay |
No cell-free enzyme/receptor binding data specifically for iRucaparib-AP6 are published. However, the rucaparib warhead has been characterized: it binds to PARP1 with a Ki of 1.4 nM and to PARP2 with a Ki of 2.0 nM. The PROTAC's binding affinity to PARP1 is expected to be similar (high nM range). For CRBN binding, the pomalidomide ligand has a Kd of 300-500 nM for CRBN. The whole PROTAC molecule likely has reduced affinity due to the linker, but it retains sufficient binding to form a ternary complex.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: Primary rat neonatal cardiomyocytes ( Tested Concentrations: 0.001 μM; 0 -20 μM; 24 hrs (hours)) induces PARP1 degradation at low concentrations[1]. 0.01μM; 0.1μM; 1μM; 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: PARP-1 levels were diminished in primary rat neonatal cardiomyocytes. Western Blot Analysis [1] Cell Types: Primary rat neonatal cardiomyocytes Tested Concentrations: 0.05 μM; 0.1 μM; 0.2 μM; 0.5 μM; 1 μM; 2 μM; 5 μM; 10 μM; 20 μM Incubation Duration: 24 hrs (hours) Experimental Results: Concentrations as low as Strong degradation of PARP1 is induced at 50 nM. Primary rat neonatal cardiomyocytes (RNCMs) are isolated from 1-2 day old rat pups and cultured in DMEM with 10% FBS. Cells are seeded in 12-well plates (2×10^5 cells/well). After 48 h, cells are treated with iRucaparib-AP6 at concentrations of 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20 uM for 24 h. Whole-cell lysates are prepared, and PARP1 levels are analyzed by western blot using anti-PARP1 antibody (dilution 1:1000). GAPDH or beta-actin is used as loading control. Degradation is quantified by densitometry, and DC50 is calculated using non-linear regression. For NAD+ depletion assays, cells are treated with 1 uM doxorubicin (DNA damage) +/- iRucaparib-AP6 (0.2-2 uM) for 6 h, then NAD+ is measured via an enzymatic cycling assay. |
| Animal Protocol |
No animal studies have been reported. A typical study would involve administering iRucaparib-AP6 to mice bearing tumor xenografts (e.g., MDA-MB-231 breast cancer) via intraperitoneal injection at doses of 5-50 mg/kg, once daily or every other day for 2-3 weeks. Tumor tissues would be collected at various time points (4-48 h after dosing) to assess PARP1 degradation by western blot. Tumor growth would be measured. In addition, plasma and tissue levels of the PROTAC would be quantified by LC-MS/MS. However, no such data are available.
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| ADME/Pharmacokinetics |
No PK data are available. As a PROTAC, iRucaparib-AP6 has a high molecular weight (approximately 850-900 Da), high LogP (estimated >5), and multiple amide bonds, which typically result in poor oral bioavailability (<5% in general for PROTACs). If administered intravenously, half-life is likely 1-3 hours in mice due to rapid clearance (hepatic metabolism and biliary excretion). The compound is a substrate for P-glycoprotein (likely). No detailed ADME studies have been published. Formulation in 10% DMSO + 10% Cremophor EL + 80% saline is typical for in vivo studies of PROTACs.
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| Toxicity/Toxicokinetics |
No toxicity data are reported. In cell culture, iRucaparib-AP6 shows no significant cytotoxicity in RNCMs at concentrations up to 5 uM (as measured by LDH release or propidium iodide staining). At 20 uM, some non-specific toxicity may occur. The compound is not expected to cause acute toxicity in animals at efficacious doses (estimated <50 mg/kg). However, off-target degradation of other proteins that contain similar PARP1-binding motifs or of CRBN neosubstrates (e.g., IKZF1, IKZF3) could lead to immunomodulatory effects, as pomalidomide is known to degrade these transcription factors. This could result in immune-related toxicities (e.g., lymphopenia, teratogenicity). Long-term toxicity studies have not been conducted.
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| References | |
| Additional Infomation |
iRucaparib-AP6 is a research-grade PROTAC tool for studying PARP1 biology and for validating the therapeutic potential of PARP1 degradation in cancer, particularly in PARP inhibitor-resistant tumors. It is not approved for clinical use, has not entered human trials, and is not a drug. The compound is protected by patents (e.g., WO2020123286A1) from academic institutions. It is available from chemical suppliers for research only. This compound represents a new class of PARP-directed therapy that goes beyond inhibition. It is sometimes referred to as a "PARP1 degrader" and is used in combination with DNA-damaging agents. It is not to be confused with other PARP1 PROTACs (e.g., XZ-1, PZ-1).
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| Molecular Formula |
C46H55FN6O11
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|---|---|
| Molecular Weight |
886.9609
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| Exact Mass |
886.391
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| CAS # |
2410557-00-3
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| PubChem CID |
138857977
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
2.5
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
26
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| Heavy Atom Count |
64
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| Complexity |
1540
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| Defined Atom Stereocenter Count |
0
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| SMILES |
FC1=C([H])C2C(N([H])C([H])([H])C([H])([H])C3=C(C4C([H])=C([H])C(C([H])([H])N(C([H])([H])[H])C([H])([H])C([H])([H])OC([H])([H])C([H])([H])OC([H])([H])C([H])([H])OC([H])([H])C([H])([H])OC([H])([H])C([H])([H])OC([H])([H])C([H])([H])OC([H])([H])C([H])([H])N([H])C5=C([H])C([H])=C([H])C6C(N(C(C=65)=O)C5([H])C(N([H])C(C([H])([H])C5([H])[H])=O)=O)=O)=C([H])C=4[H])N([H])C(=C1[H])C3=2)=O
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| InChi Key |
YHMDCINUVWULST-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C46H55FN6O11/c1-52(29-30-5-7-31(8-6-30)42-33-11-12-49-43(55)35-27-32(47)28-37(50-42)40(33)35)14-16-60-18-20-62-22-24-64-26-25-63-23-21-61-19-17-59-15-13-48-36-4-2-3-34-41(36)46(58)53(45(34)57)38-9-10-39(54)51-44(38)56/h2-8,27-28,38,48,50H,9-26,29H2,1H3,(H,49,55)(H,51,54,56)
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| Chemical Name |
2-(2,6-dioxopiperidin-3-yl)-4-[2-[2-[2-[2-[2-[2-[2-[[4-(6-fluoro-9-oxo-3,10-diazatricyclo[6.4.1.04,13]trideca-1,4,6,8(13)-tetraen-2-yl)phenyl]methyl-methylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethylamino]isoindole-1,3-dione
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~56.37 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.82 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.1274 mL | 5.6372 mL | 11.2745 mL | |
| 5 mM | 0.2255 mL | 1.1274 mL | 2.2549 mL | |
| 10 mM | 0.1127 mL | 0.5637 mL | 1.1274 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.
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