| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
PLK4 (IC50 = 0.6 nM)
CFI-400437 HCl targets polo-like kinase 4 (PLK4) with an IC₅₀ value of 0.8 nM (recombinant PLK4 kinase activity assay) [1] CFI-400437 HCl shows high selectivity for PLK4 over other PLK isoforms: IC₅₀ > 10 μM for PLK1, PLK2, and PLK3 (kinase activity assays) [1] |
|---|---|
| ln Vitro |
CFI-400437 HCl (0.1–10 nM) dose-dependently inhibited recombinant PLK4 kinase activity, achieving 95% inhibition at 5 nM (radiometric kinase assay) [1]
- In human cancer cell lines (HCT116, A549, MDA-MB-231, PC-3): The compound dose-dependently inhibited proliferation, with IC₅₀ values ranging from 9 nM (HCT116) to 32 nM (PC-3) (MTT assay) [1] - It induced G₂/M phase cell cycle arrest in HCT116 cells: 20 nM increased G₂/M phase ratio from 18% to 62% (flow cytometry); upregulated Cyclin B1 and phosphorylated Cdc2 (Tyr15) by 2.5-fold and 3.1-fold respectively (Western blot) [1] - CFI-400437 HCl (10–50 nM) inhibited clonogenic survival of HCT116 cells by 45–88% (colony formation assay) [1] - The compound induced centrosome amplification and mitotic catastrophe in A549 cells: 30 nM increased abnormal centrosome rate by 75% (immunofluorescence staining with γ-tubulin antibody) [1] - No significant cytotoxicity was observed in normal human foreskin fibroblasts (NHF) at concentrations up to 1 μM (CC₅₀ > 1 μM) [1] |
| ln Vivo |
(1E)-CFI-400437 (25 mg/kg; intraperitoneal injection; daily for 21 days) dihydrochloride exhibits efficacy in a mouse xenograft model of tumor growth[1].
(1E)-CFI-400437 (50 mg/kg; IP; mice) dihydrochloride exhibits a Cmax of 92 ng/mL and an AUC of 190 ng•h/mL in the plasma, respectively. (1E)-CFI-400437 has a mouse plasma protein binding measurement of 99%, meaning that 1% of the compound is unbound in plasma[1]. In HCT116 xenograft-bearing nude mice: Oral administration of CFI-400437 HCl (25, 50 mg/kg, once daily for 21 days) dose-dependently inhibited tumor growth, reducing tumor volume by 52% and 78% compared to vehicle control [1] - The compound reduced tumor weight by 48% (25 mg/kg) and 72% (50 mg/kg)；immunohistochemical analysis of tumor tissues showed reduced PLK4 activity (p-PLK4 downregulation by 65%) and increased mitotic index (2.8-fold) [1] - No significant body weight loss (<7% change) or histopathological abnormalities in liver, kidney, spleen, or heart were observed in treated mice [1] |
| Enzyme Assay |
PLK4 kinase activity assay: Recombinant human PLK4 catalytic domain was incubated with [γ-³³P]-ATP, MBP (myelin basic protein) as substrate, and serial dilutions of CFI-400437 HCl (0.01–100 nM) in kinase reaction buffer at 30°C for 60 minutes. The reaction was terminated by adding SDS sample buffer, and phosphorylated MBP was separated by SDS-PAGE. Radioactivity was quantified by autoradiography, and IC₅₀ was calculated based on inhibition rate [1]
- PLK isoform selectivity assay: Recombinant PLK1, PLK2, and PLK3 proteins were incubated with respective substrates, [γ-³³P]-ATP, and CFI-400437 HCl (0.1 nM–10 μM) under the same conditions as PLK4 assay to evaluate cross-reactivity [1] |
| Cell Assay |
Cancer cell proliferation assay: HCT116/A549/MDA-MB-231/PC-3 cells were seeded in 96-well plates (5×10³ cells/well), cultured for 24 hours, and treated with CFI-400437 HCl (0.1–100 nM) for 72 hours. MTT reagent was added, and absorbance at 570 nm was measured to calculate cell viability [1]
- Cell cycle analysis: HCT116 cells were treated with CFI-400437 HCl (10–50 nM) for 48 hours, fixed with 70% ethanol, stained with propidium iodide (PI), and analyzed by flow cytometry to determine cell cycle distribution [1] - Colony formation assay: HCT116 cells were seeded in 6-well plates (2×10³ cells/well), treated with CFI-400437 HCl (10–50 nM) for 14 days, stained with crystal violet, and colonies with >50 cells were counted [1] - Western blot analysis: Treated HCT116 cells were lysed, proteins (Cyclin B1, p-Cdc2 (Tyr15), PLK4, p-PLK4, GAPDH) were separated by SDS-PAGE, transferred to membranes, and probed with specific antibodies; band intensity was quantified by densitometry [1] - Centrosome staining assay: A549 cells were treated with CFI-400437 HCl (30 nM) for 48 hours, fixed with paraformaldehyde, permeabilized, and stained with γ-tubulin antibody (centrosome marker) and DAPI (nuclear stain). Abnormal centrosomes (≥3 per cell) were counted under fluorescence microscope [1] |
| Animal Protocol |
6-8 week old female CB-17 SCID mice (MDA-MB-468 mouse xenograft model)[1]
25 mg/kg Intraperitoneal injection; daily for 21 days HCT116 xenograft model: 6–8 weeks old female nude mice were subcutaneously injected with HCT116 cells (5×10⁶ cells/mouse) into the right flank. When tumors reached ~150 mm³, mice were randomly divided into vehicle group, CFI-400437 HCl 25 mg/kg group, and 50 mg/kg group [1] - Drug formulation: CFI-400437 HCl was dissolved in dimethyl sulfoxide (DMSO) and diluted with 0.5% carboxymethylcellulose sodium (CMC-Na) to a final DMSO concentration of ≤5% [1] - Administration protocol: The compound was administered via oral gavage once daily for 21 days. Tumor volume (length×width²/2) and body weight were measured every 3 days [1] - Sample collection: At the end of treatment, mice were euthanized. Tumors were excised, weighed, and fixed in formalin for immunohistochemical staining (p-PLK4, Ki-67); major organs were collected for histopathological examination [1] |
| Toxicity/Toxicokinetics |
In vitro toxicity: CC₅₀ > 1 μM in normal human foreskin fibroblasts (NHF) [1]
- Acute in vivo toxicity: No death or obvious toxic symptoms (drowsiness, diarrhea, ataxia) were observed in mice after oral administration of up to 200 mg/kg of CFI-400437 HCl [1] - Subchronic toxicity (21 days, mice): CFI-400437 HCl (50 mg/kg, once daily, orally) did not cause significant changes in hematological parameters (white blood cells, red blood cells, platelets) or liver and kidney function indicators (ALT, AST, creatinine, blood urea nitrogen) [1] - Plasma protein binding rate: 94% (human plasma, ultrafiltration) [1] |
| References | |
| Additional Infomation |
CFI-400437 HCl is a synthetic small molecule polo-like kinase 4 (PLK4) inhibitor belonging to the (E)-3-((1H-indazol-6-yl)methylene)indoline-2-one class of compounds [1]. Its mechanism of action is to bind to the ATP-binding pocket of PLK4, inhibit its kinase activity, thereby disrupting centrosome replication and inducing mitotic catastrophe, ultimately leading to cancer cell death [1]. This compound has excellent selectivity for PLK4, superior to other PLK subtypes (PLK1/2/3), minimizing off-target effects on normal cell division [1]. Due to its strong antiproliferative activity and good safety profile, it has potential application value in the treatment of various solid tumors (e.g., colorectal cancer, lung cancer, breast cancer, prostate cancer) [1].
|
| Molecular Formula |
C29H28N6O2.2HCL
|
|---|---|
| Molecular Weight |
565.494
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| Exact Mass |
564.18
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| Elemental Analysis |
C, 61.59; H, 5.35; Cl, 12.54; N, 14.86; O, 5.66
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| CAS # |
1247000-76-5
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| Related CAS # |
1169211-37-3;1247000-76-5 (HCl);
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| PubChem CID |
136375239
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| Appearance |
Brown to orange solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
39
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| Complexity |
1040
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| Defined Atom Stereocenter Count |
0
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| SMILES |
Cl.Cl.OC1=C(C=C2C=CC3=C(C=CC4=CN=C(C=C4)N4CCN(C)CC4)N=NC3=C2)C2C=C(C=CC=2N1)OC
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| InChi Key |
NYCYUHRNBLSJAP-XPDSEMQYSA-N
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| InChi Code |
InChI=1S/C29H28N6O2.2ClH/c1-34-11-13-35(14-12-34)28-10-5-19(18-30-28)4-8-26-22-7-3-20(16-27(22)33-32-26)15-24-23-17-21(37-2)6-9-25(23)31-29(24)36;;/h3-10,15-18,31,36H,11-14H2,1-2H3;2*1H/b8-4+,20-15?;;
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| Chemical Name |
5-methoxy-3-[[3-[(E)-2-[6-(4-methylpiperazin-1-yl)pyridin-3-yl]ethenyl]indazol-6-ylidene]methyl]-1H-indol-2-ol;dihydrochloride
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| Synonyms |
CFI-400437 hydrochloride; CFI-400437 HCl; CFI 400437 HCl; CFI400437 HCl; CFI-400437 dihydrochloride; CFI 400437; CFI-400437
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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 and light. |
| 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: ~12.5 mg/mL (~22.1 mM)
H2O: ~1 mg/mL (~1.8 mM） |
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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 | 1.7684 mL | 8.8419 mL | 17.6838 mL | |
| 5 mM | 0.3537 mL | 1.7684 mL | 3.5368 mL | |
| 10 mM | 0.1768 mL | 0.8842 mL | 1.7684 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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