| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
BRD4 BD1 0.059 μM (IC50) PLK1 0.127 μM (IC50) BRDT BD1 0.245 μM (IC50)
BRD4 (bromodomain 4, specifically the first bromodomain, BD1) and PLK1 (polo‑like kinase 1). The compound also inhibits BRDT-BD1 (the bromodomain of bromodomain testis‑specific protein). |
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| ln Vitro |
In cell‑free enzymatic assays, PLK1/BRD4-IN-3 inhibits BRD4-BD1 with an IC₅0 of 0.059 uM (59 nM), PLK1 with an IC₅0 of 0.127 uM (127 nM), and BRDT-BD1 with an IC₅0 of 0.245 uM (245 nM). This balanced dual inhibition profile demonstrates high potency against both targets.
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| ln Vivo |
This entry is not available. As a dual inhibitor, PLK1/BRD4-IN-3 is expected to induce both G2/M cell cycle arrest (due to PLK1 inhibition) and downregulation of oncogenes such as MYC (due to BRD4 inhibition). The combination of these effects should lead to synergistic anti‑proliferative activity in cancer cells.
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| Enzyme Assay |
A general cell‑free protocol for BRD4 inhibition: A TR‑FRET assay is used. Recombinant BRD4(BD1) protein (2 nM) is incubated with a biotinylated acetylated histone H4 peptide (50 nM) in assay buffer (50 mM HEPES, pH 7.5, 150 mM NaCl, 1 mM DTT, 0.01% Tween‑20, 0.1% BSA) with varying concentrations of PLK1/BRD4-IN-3 (0.001‑10 uM). Europium‑labeled anti‑GST antibody (1 nM) and streptavidin‑XL665 (20 nM) are added. After 1 hour, the FRET signal is measured at 665 nm. The IC₅0 is calculated. For PLK1 inhibition, a radiometric kinase assay is used. Recombinant PLK1 (5 nM) is incubated with a biotinylated peptide substrate (1 uM) and 10 uM ATP with varying concentrations of PLK1/BRD4-IN-3. The reaction is incubated at 30degC for 30 minutes, and the phosphorylated substrate is detected by TR‑FRET.
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| Cell Assay |
A general cellular protocol for PLK1/BRD4 inhibitors: Cancer cells (e.g., HeLa, MV4‑11, or A549) are seeded in 96‑well plates and treated with serial dilutions of PLK1/BRD4-IN-3 (0.001‑10 uM) for 72 hours. Cell viability is measured using CellTiter‑Glo or MTT assay. For cell cycle analysis, cells are treated for 24 hours, fixed, stained with propidium iodide, and analyzed by flow cytometry. Mitotic index is assessed by staining with anti‑phospho‑histone H3 (Ser10) antibody. For BRD4 inhibition, cells are treated for 6‑24 hours, and MYC mRNA levels are measured by qRT‑PCR, and MYC protein levels are measured by Western blot. For PLK1 inhibition, cells are treated for 24 hours, and phospho‑PLK1 (Thr210) and phospho‑Cdc25C (Ser216) levels are measured by Western blot.
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| Animal Protocol |
A general animal protocol for PLK1/BRD4-IN-3: Female NOD/SCID mice are subcutaneously implanted with 5×10⁶ MV4‑11 leukemia cells or HeLa cervical cancer cells. When tumors reach ~150 mm3, mice are randomized into treatment groups (n=8/group). PLK1/BRD4-IN-3 is formulated in a suitable vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween‑80, 45% saline) and administered via oral gavage at doses of 10, 30, and 100 mg/kg once daily for 21 days. Tumor volume is measured twice weekly. At study end, tumors are harvested for Western blot (MYC, phospho‑histone H3, cleaved PARP), IHC (Ki‑67, phospho‑histone H3), and qRT‑PCR (MYC). A PK/PD study is performed to correlate plasma exposure with target inhibition.
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| ADME/Pharmacokinetics |
General PK protocol for PLK1/BRD4-IN-3: Male Sprague‑Dawley rats are administered PLK1/BRD4-IN-3 via oral gavage (PO, 10 mg/kg) and intravenous (IV, 2 mg/kg). Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours. Plasma concentrations are quantified by LC‑MS/MS. PK parameters (Cmax, Tmax, AUC, t½, clearance, Vd, and oral bioavailability) are calculated. The compound has a molecular weight of 561.72 and is soluble in DMSO at 16.67 mg/mL.
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| Toxicity/Toxicokinetics |
General toxicity protocol for PLK1/BRD4-IN-3: A 14‑day repeat‑dose oral toxicity study is performed in ICR mice. PLK1/BRD4-IN-3 is administered via oral gavage at doses of 10, 30, and 100 mg/kg/day for 14 days. Parameters include clinical signs, body weight, food consumption, hematology (complete blood count with differential, platelet count), serum chemistry (ALT, AST, BUN, creatinine), and histopathology of bone marrow, gastrointestinal tract, liver, and kidney. Special attention is paid to the bone marrow (myelosuppression) and GI tract (gastrointestinal toxicity), which are on‑target effects of PLK1 and BRD4 inhibition.
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| References | |
| Additional Infomation |
PLK1/BRD4-IN-3 was developed through a structure‑guided design and optimization campaign that produced a series of potent and selective dual BRD4/PLK1 inhibitors (including PLK1/BRD4-IN-2 and PLK1/BRD4-IN-3). The dual inhibitor strategy is designed to overcome resistance mechanisms associated with single‑target therapy and to provide improved anti‑tumor efficacy. The compound has the CAS number 2251709-91-6. The molecular formula is C31H43N₇O3, and the molecular weight is 561.72 g/mol.
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| Molecular Formula |
C31H43N7O3
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|---|---|
| Molecular Weight |
561.72
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| Exact Mass |
561.343
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| CAS # |
2251709-91-6
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| PubChem CID |
145976646
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
41
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| Complexity |
902
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| Defined Atom Stereocenter Count |
1
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| SMILES |
C[C@H]1C(=O)N(C2=CN=C(N=C2N1C3CCCC3)NC4=C(C=C(C=C4)C(=O)NC5CCN(CC5)C)OC6CCCC6)C
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| InChi Key |
HTPNGOWITIWAHM-FQEVSTJZSA-N
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| InChi Code |
InChI=1S/C31H43N7O3/c1-20-30(40)37(3)26-19-32-31(35-28(26)38(20)23-8-4-5-9-23)34-25-13-12-21(18-27(25)41-24-10-6-7-11-24)29(39)33-22-14-16-36(2)17-15-22/h12-13,18-20,22-24H,4-11,14-17H2,1-3H3,(H,33,39)(H,32,34,35)/t20-/m0/s1
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| Chemical Name |
4-[[(7S)-8-cyclopentyl-5,7-dimethyl-6-oxo-7H-pteridin-2-yl]amino]-3-cyclopentyloxy-N-(1-methylpiperidin-4-yl)benzamide
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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 |
| 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) |
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
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7802 mL | 8.9012 mL | 17.8025 mL | |
| 5 mM | 0.3560 mL | 1.7802 mL | 3.5605 mL | |
| 10 mM | 0.1780 mL | 0.8901 mL | 1.7802 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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