| Size | Price | Stock | Qty |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
(R)-Filanesib targets the kinesin spindle protein (KSP, also known as Eg5), a mitotic kinesin that is essential for the formation of the bipolar mitotic spindle. It acts as a potent inhibitor of KSP, with an IC50 of 6 nM. By inhibiting KSP, the compound prevents the separation of centrosomes and the formation of a functional mitotic spindle, leading to mitotic arrest. This results in the activation of the spindle assembly checkpoint and ultimately induces apoptosis in dividing cells.
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| ln Vitro |
Filanesib (ARRY520; ARRY-520) continues to function in cell lines resistant to multiple drugs. Filanesib (ARRY-520) has EC50s of 3.7, 14, and 4.2 nM for inhibiting the proliferation of HCT-15, NCI/ADR-RES, and K562/ADR cells, respectively. Most cells in mitosis are blocked by filanesib (ARRY-520) (10 nM) with the monopolar spindle structure characteristic of KSP inhibition[1]. In four cells, increased phosphorylation of histone H3 (pHH3) and accumulation of cyclin B1 indicate that filanesib (ARRY-520) (10 nM) induces mitotic arrest[2]. The cytotoxic effects of Paclitaxel and Filanesib (ARRY-520) on Type I and II cells are identical. For Type II EOC cells, the GI50 at 48 hours is 0.0015 μM for ARRY-520. The GI50 for ARRY-520 at 48 hours is greater than 3 μM in Type I EOC cells[3]. In OCI-AML3 cells, filanesib (ARRY-520) at 1 nM causes a substantial G2M cell cycle block within 24 hours[4].
In vitro studies have shown that (R)-Filanesib is a potent inhibitor of KSP with an IC50 of approximately 6 nM. It induces mitotic arrest and apoptosis in proliferating cancer cells. The compound has demonstrated activity against various cancer cell lines, with particular potency in hematologic malignancies such as multiple myeloma. By inhibiting KSP-mediated spindle formation, it suppresses cell proliferation. The R-enantiomer is the active form of the compound, with the S-enantiomer showing significantly reduced activity. |
| ln Vivo |
Filanesib (ARRY520; ARRY-520) (10, 15, 20, 30 mg/kg, i.p.) is more effective than paclitaxel in mice with subcutaneous HT-29, HCT-116, MDA-MB-231, and A2780 xenografts, as well as active in UISO-BCA-1 xenografts. In both the DU145 prostate xenograft model and the androgen receptor-negative PC-3 prostate cancer xenograft model, ARRY-520 outperforms docetaxel[1]. Tumor xenografts from RPMI 8226 are especially responsive to low dosages of ARRY-520 (12.5 mg/kg, i.p.)[2]. In HL60 and MV4-11 xenografts of SCID mice, ARRY-520 dramatically inhibits tumor growth at doses of 27 mg/kg and 20 mg/kg, respectively[4].
In vivo, (R)-Filanesib has shown antitumor efficacy in preclinical models of multiple myeloma and other cancers. It has been evaluated in clinical trials for the treatment of hematologic malignancies. The compound induces mitotic arrest and apoptosis in tumor tissues, leading to tumor growth inhibition. Its in vivo activity is dose-dependent, and it has been studied both as a monotherapy and in combination with other anticancer agents. The compound's efficacy in preclinical models has supported its clinical development for oncology indications. |
| Enzyme Assay |
The in vitro enzyme assay for (R)-Filanesib involves measuring the inhibition of KSP ATPase activity. Purified KSP protein is incubated with ATP and a microtubule substrate. The ATPase activity of KSP is measured by detecting the release of inorganic phosphate using a colorimetric or fluorometric assay. Varying concentrations of (R)-Filanesib are added to the reaction, and the inhibition of ATPase activity is determined. The IC50 of 6 nM is calculated from the dose-response curve. Data are analyzed using nonlinear regression models.
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| Cell Assay |
Filanesib (ARRY-520) is applied to exponentially growing cells (0.4×106/mL) for a maximum of 48 hours. For a maximum of 96 hours, Filanesib (ARRY-520), ABT-737, or both are incubated with HL-60 and HL-60Bcl-2 cells (0.4×106/mL). The control agent in this case is DMSO. The Annexin-V-FLUOS Staining Kit is used in flow cytometry measurements of phosphatidyl serine to estimate apoptosis. Simultaneous assessment of membrane integrity is conducted with 7-amino-actinomycin D (7-AAD). For one hour at 37°C, cells are loaded with CMXRos (300 nM) and MitoTracker Green (500 nM) in order to measure changes in the mitochondrial membrane potential (MMP). Next, CMXRos retention is measured and the loss of MMP is evaluated while simultaneously correcting for mitochondrial mass.
In vitro cellular assays for (R)-Filanesib are conducted using cancer cell lines, particularly those derived from hematologic malignancies such as multiple myeloma (e.g., RPMI-8226, U266). Cells are plated in multi-well plates and treated with varying concentrations of (R)-Filanesib. Cell viability is assessed using standard assays such as MTT, CellTiter-Glo, or flow cytometry-based viability staining. Mitotic arrest is evaluated by measuring the mitotic index (phospho-histone H3 staining) using flow cytometry or immunocytochemistry. Apoptosis is assessed by measuring caspase activity or Annexin V staining. |
| Animal Protocol |
Tumor xenografts placed subcutaneously are permitted to expand to a volume of 250–350 mm3. Based on the size of their tumors, the mice are randomly assigned to groups of three to four, and each group receives a single intraperitoneal dose of filanesib (ARRY-520). The mice are put to death by CO2 inhalation at different intervals following the drug's administration, and the tumors are removed and put in 10% neutral buffered formalin. Standard procedures are followed to process and paraffin embed the formalin-fixed tumors. TUNEL stain is used to analyze apoptosis, and α-tubulin staining is used to analyze spindle morphology in tumor sections. For every sample, three ×40 fields are counted to identify monopolar/abnormal spindles and TUNEL positive (apoptotic) cells, which are then analyzed using algorithms built into the ImagePro software.
In vivo animal studies for (R)-Filanesib are performed in mouse xenograft models of human cancer, particularly multiple myeloma. Immunocompromised mice are implanted with tumor cells subcutaneously. When tumors reach a certain size, animals are treated with (R)-Filanesib via intraperitoneal or oral administration. Tumor growth is monitored by caliper measurements. At endpoint, tumors are collected for histopathological analysis, and biomarkers of mitotic arrest and apoptosis (e.g., phospho-histone H3, cleaved caspase-3) are assessed. Standard study designs with vehicle control and positive control groups are employed. |
| ADME/Pharmacokinetics |
Pharmacokinetic data for (R)-Filanesib are not extensively reported in the available literature. The compound has a molecular weight of 420.48 g/mol and a molecular formula of C20H22F2N4O2S. It is a small molecule with properties suitable for oral bioavailability. In preclinical studies, it has been administered via intraperitoneal or oral routes. Specific pharmacokinetic parameters such as half-life, Cmax, and bioavailability are not provided in the available sources. The compound's pharmacokinetic profile would be consistent with other small-molecule kinase inhibitors.
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| Toxicity/Toxicokinetics |
Toxicology data for (R)-Filanesib are not extensively reported. As a KSP inhibitor, the compound's primary toxicity would be related to its effects on rapidly dividing cells, including bone marrow, gastrointestinal epithelium, and hair follicles. Common toxicities associated with mitotic inhibitors include myelosuppression, gastrointestinal disturbances, and alopecia. Preclinical toxicology studies have been conducted to support clinical development. Specific toxicity data, including LD50 values and organ toxicity profiles, are not available in the public domain.
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| References | |
| Additional Infomation |
(R)-Filanesib (ARRY-520 R enantiomer) is a potent inhibitor of kinesin spindle protein (KSP) with an IC50 of 6 nM. It induces mitotic arrest and apoptosis in proliferating cancer cells. The compound has been investigated for the treatment of hematologic malignancies, particularly multiple myeloma. It has been evaluated in clinical trials as an anticancer agent. (R)-Filanesib is the active R-enantiomer of filanesib and is a valuable research tool for studying mitosis and KSP biology. It is for research use only.
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| Molecular Formula |
C20H22F2N4O2S
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|---|---|
| Molecular Weight |
420.476089954376
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| Exact Mass |
420.143
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| CAS # |
885060-08-2
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| Related CAS # |
Filanesib;885060-09-3
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| PubChem CID |
46189838
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| Appearance |
White to off-white solid
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| LogP |
3.954
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
29
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| Complexity |
605
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| Defined Atom Stereocenter Count |
1
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| SMILES |
C([C@@]1(SC(C2C=C(F)C=CC=2F)=NN1C(=O)N(C)OC)C1C=CC=CC=1)CCN
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| InChi Key |
LLXISKGBWFTGEI-HXUWFJFHSA-N
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| InChi Code |
InChI=1S/C20H22F2N4O2S/c1-25(28-2)19(27)26-20(11-6-12-23,14-7-4-3-5-8-14)29-18(24-26)16-13-15(21)9-10-17(16)22/h3-5,7-10,13H,6,11-12,23H2,1-2H3/t20-/m1/s1
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| Chemical Name |
(2R)-2-(3-aminopropyl)-5-(2,5-difluorophenyl)-N-methoxy-N-methyl-2-phenyl-1,3,4-thiadiazole-3-carboxamide
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| Synonyms |
(R)-ARRY-520; (R)-Filanesib
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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) |
DMSO: ≥ 100 mg/mL (~237.8 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3.75 mg/mL (8.92 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 37.5 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: ≥ 3.75 mg/mL (8.92 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 37.5 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 | 2.3782 mL | 11.8912 mL | 23.7823 mL | |
| 5 mM | 0.4756 mL | 2.3782 mL | 4.7565 mL | |
| 10 mM | 0.2378 mL | 1.1891 mL | 2.3782 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.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT02384083 | Completed | Drug: Filanesib, pomalidomide and dexamethasone |
Multiple Myeloma | PETHEMA Foundation | September 2015 | Phase 1 Phase 2 |
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