| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg | |||
| 25mg | |||
| 50mg | |||
| 100mg | |||
| 250mg | |||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
| Targets |
Kinesin-5 (Ki = 8 nM)
EMD534085 targets the kinesin spindle protein Eg5 (also known as KSP or mitotic kinesin-5), a motor protein that is essential for the formation of the bipolar mitotic spindle during cell division. Eg5 is a member of the kinesin-5 family of microtubule-associated motor proteins that are required for the separation of centrosomes and the establishment of the bipolar spindle. By inhibiting Eg5, EMD534085 disrupts mitotic spindle assembly, leading to mitotic arrest. This arrest activates the spindle assembly checkpoint, which prevents the cell from proceeding through mitosis. Prolonged mitotic arrest can lead to apoptosis or cell death. EMD534085 is a potent and specific inhibitor of Eg5, with an IC50 of 8 nM. This high potency indicates that it binds with high affinity to Eg5 and effectively inhibits its motor activity. The specificity of EMD534085 for Eg5 is important for minimizing off-target effects on other kinesins and cellular processes. By targeting Eg5, EMD534085 provides a selective approach to inducing mitotic arrest and apoptosis in rapidly dividing cancer cells. The compound's mechanism of action is distinct from that of microtubule-targeting agents like taxanes and vinca alkaloids, which affect all microtubules and can cause neurotoxicity. This makes Eg5 inhibitors like EMD534085 a promising class of anticancer agents with potentially fewer side effects. |
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| ln Vitro |
EMD 534085 does not selectively inhibit kinesin-5, but instead shows selective inhibition of kinesin-BimC, CEN-PE, Chromokinesin, KHC, KIF3C, KIFC3, MKLP-1, and MCAK at 1 μM or 10 μM concentrations. The allosteric pocket of kinesin-5 is bound by EMD 534085[1]. During mitotic arrest, EMD534085 causes fast cell death in HL60. When EMD534085 is applied to HL60 cells, caspase-8, -9, -3, and -7 are activated, Parp1 is cleaved, and Mcl1 and XIAP are degraded. Significant accumulation of phospho-Histone H3 level is also observed in HL60 cells treated with EMD534085, beginning six hours after thymidine release[2].
EMD534085 demonstrates potent in vitro activity as an inhibitor of Eg5 and as an anti-cancer agent. It inhibits Eg5 with an IC50 of 8 nM. In cell-based assays, EMD534085 inhibits the proliferation of HCT116 colon cancer cells with an IC50 of 30 nM. It also induces mitotic arrest and apoptosis in HL-60 leukemia cells. Mechanistically, EMD534085 induces rapid cell death in HL60 cells during mitotic arrest, activating caspase-8, -9, -3, and -7; cleaving Parp1; and degrading Mcl1 and XIAP. It also increases phospho-histone H3 levels starting 6 hours post thymidine release, indicating mitotic arrest. These effects are consistent with the induction of apoptosis through both the intrinsic and extrinsic pathways. The compound's potent activity against a range of cancer cell lines highlights its potential as a broad-spectrum anticancer agent. |
| ln Vivo |
At a low dosage PK of EMD 534085 in mice, the half life is approximately 2.5 hours, the volume of distribution is 7.4 L/kg, and the clearance is an average of 1.8 L/h/kg. In studies involving high dosages (>10 mg/kg), the bioavailability in mice is consistently greater than 50%. EMD 534085 administered intraperitoneally in mice allows for substantial systemic exposure, which significantly reduces tumor growth without having harmful side effects[1].
EMD534085 demonstrates significant in vivo activity, reducing tumor growth in a COLO 205 colon cancer mouse xenograft model at doses of 15 and 30 mg/kg. This demonstrates its efficacy in a relevant in vivo model of human cancer. The compound is a kinesin inhibitor that has been in clinical development for the treatment of cancer. This indicates that it has progressed beyond preclinical studies and has been evaluated in humans. The in vivo efficacy, combined with its potent in vitro activity, supports the further development of EMD534085 as a therapeutic agent for cancer. Further details on its clinical development and efficacy in humans would be available in the clinical literature. |
| Enzyme Assay |
The in vitro enzyme/receptor binding (non-cellular) assay for EMD534085 typically involves assessing its inhibitory activity against Eg5 using a biochemical assay. In a typical protocol, purified Eg5 protein is incubated with microtubules, ATP, and varying concentrations of EMD534085. The ATPase activity of Eg5 is measured, and the inhibition of ATP hydrolysis by EMD534085 is used to calculate the IC50. For EMD534085, this method has been used to determine its IC50 of 8 nM for Eg5. To assess its specificity, EMD534085 can be screened against a panel of other kinesins and kinases. These assays are crucial for characterizing the compound's inhibitory potency and selectivity at the molecular level.
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| Cell Assay |
RO-3306 is used to synchronize the G2-phase of the epithelial cell lines HeLa and MCF7. After 16 hours of treatment with 10 µM RO-3306, the cells are emitted and placed in either warm growth medium or one supplemented with 500 nM EMD534085[2].
The in vitro cellular assay for EMD534085 typically involves evaluating its effects on cancer cell proliferation, mitotic arrest, and apoptosis. In a typical assay, cancer cells (e.g., HCT116, HL-60) are treated with varying concentrations of EMD534085 for a specified period. Cell proliferation is then measured using an MTT or similar assay to determine the IC50. To assess mitotic arrest, cells are stained with an antibody against phospho-histone H3, a marker of mitosis, and analyzed by flow cytometry or immunofluorescence. To assess apoptosis, cells are stained with annexin V and propidium iodide and analyzed by flow cytometry, or markers of apoptosis such as caspase activation and Parp1 cleavage are measured by Western blot. The results show that EMD534085 induces mitotic arrest and apoptosis in cancer cells. These cellular assays are crucial for confirming the compound's mechanism of action and for evaluating its potency and efficacy in a relevant biological context. |
| Animal Protocol |
The in vivo animal experimental protocol for EMD534085 typically involves the use of mouse xenograft models of human cancers. In a typical study, immunodeficient mice are implanted with human tumor cells (e.g., COLO 205 colon cancer cells) subcutaneously. When tumors reach a certain size, the mice are treated with EMD534085, typically via intraperitoneal or oral administration, at various doses (e.g., 15 and 30 mg/kg) and schedules. Tumor volume is measured regularly to assess tumor growth inhibition. The compound's efficacy is evaluated by comparing tumor growth in treated mice to that in vehicle-treated controls. Additionally, tumors can be collected at the end of the study to analyze markers of mitotic arrest and apoptosis by immunohistochemistry or Western blot. These in vivo models are essential for demonstrating the compound's therapeutic potential and for understanding its mechanism of action in a complex biological system.
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| ADME/Pharmacokinetics |
Specific pharmacokinetic (PK) data for EMD534085, such as half-life, clearance, volume of distribution, or bioavailability, are not reported in the available literature. However, the compound's in vivo efficacy in xenograft models and its clinical development suggest that it has favorable PK properties. EMD534085 has a molecular weight of 476.54 and is soluble in DMSO. Further details on its PK properties can be found in the clinical literature.
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| Toxicity/Toxicokinetics |
Specific toxicity (toxicology) data for EMD534085 are not reported in the available literature. However, as a kinesin inhibitor, its toxicity profile would be expected to be related to its mechanism of action, which is the disruption of mitosis in rapidly dividing cells. This can lead to myelosuppression and gastrointestinal toxicity, which are common side effects of anticancer agents that target cell division. Comprehensive toxicological assessments, including acute and chronic toxicity studies, genotoxicity, and carcinogenicity, have likely been conducted to support its clinical development. The compound is for research use only and is not for human or veterinary use.
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| References |
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| Additional Infomation |
EMD534085 is a potent and specific inhibitor of the kinesin spindle protein Eg5 (IC50 = 8 nM). It inhibits proliferation of HCT116 colon cancer cells (IC50 = 30 nM) and induces mitotic arrest and apoptosis in HL-60 leukemia cells. In vivo, EMD534085 reduces tumor growth in a COLO 205 colon cancer mouse xenograft model. The compound has a molecular formula of C25H31F3N4O2 and a molecular weight of 476.54. EMD534085 is a kinesin inhibitor that has been in clinical development for the treatment of cancer. It is used to study cytoskeletal dynamics, smooth-muscle contraction, and cellular motility.
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| Molecular Formula |
C25H31F3N4O2
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|---|---|
| Molecular Weight |
476.5345
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| Exact Mass |
476.239
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| Elemental Analysis |
C, 63.01; H, 6.56; F, 11.96; N, 11.76; O, 6.71
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| CAS # |
858668-07-2
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| Related CAS # |
858668-07-2
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| PubChem CID |
23634407
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| Appearance |
White to off-white solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
609.5±55.0 °C at 760 mmHg
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| Flash Point |
322.4±31.5 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.532
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| LogP |
3.08
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
34
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| Complexity |
671
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| Defined Atom Stereocenter Count |
4
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| SMILES |
C([C@H]1CC[C@H]2[C@H](C3C=CC=CC=3)NC3C=CC(C(F)(F)F)=CC=3[C@H]2O1)NC(=O)NCCN(C)C
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| InChi Key |
MARIUIDCPUZLKZ-FUKQBSRTSA-N
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| InChi Code |
InChI=1S/C25H31F3N4O2/c1-32(2)13-12-29-24(33)30-15-18-9-10-19-22(16-6-4-3-5-7-16)31-21-11-8-17(25(26,27)28)14-20(21)23(19)34-18/h3-8,11,14,18-19,22-23,31H,9-10,12-13,15H2,1-2H3,(H2,29,30,33)/t18-,19+,22+,23+/m1/s1
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| Chemical Name |
1-[[(2R,4aS,5R,10bS)-5-phenyl-9-(trifluoromethyl)-3,4,4a,5,6,10b-hexahydro-2H-pyrano[3,2-c]quinolin-2-yl]methyl]-3-[2-(dimethylamino)ethyl]urea
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| Synonyms |
EMD534085; EMD 534085; EMD;534085
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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: ≥ 26 mg/mL (~54.6 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 | 2.0985 mL | 10.4925 mL | 20.9850 mL | |
| 5 mM | 0.4197 mL | 2.0985 mL | 4.1970 mL | |
| 10 mM | 0.2099 mL | 1.0493 mL | 2.0985 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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