| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
| Targets |
Tubulin inhibitor 8 targets tubulin, the protein that polymerizes to form microtubules. Microtubules are dynamic cytoskeletal structures that play a critical role in various cellular processes, including cell division, intracellular transport, and cell shape maintenance. During mitosis, microtubules form the mitotic spindle, which is responsible for segregating chromosomes into daughter cells. Tubulin inhibitors bind to tubulin and prevent its polymerization into microtubules, thereby disrupting the mitotic spindle and arresting cells in the G2/M phase of the cell cycle. This leads to cell cycle arrest and ultimately apoptosis. Tubulin inhibitor 8 is a potent inhibitor of tubulin polymerization, with an IC₅₀ of 0.73 μM. Its ability to inhibit tubulin polymerization is the basis for its antiproliferative activity against cancer cells. The compound is a member of the phenoxazine class of compounds.
|
|---|---|
| ln Vitro |
Tubulin inhibitor 8 prevents mitosis by stopping cells in the G2/M phase, as an example of a typical histogram would illustrate [1]. With an IC50 of 15, 6, 8, 2, 8, 6, and 1, respectively, tubulin inhibitor 8 shown good anti-proliferative effectiveness against NCIH460, SKOV3, BT549, 451LU, SW480, COLO-205, and DLD-1 tumor cell lines [1].
In vitro, Tubulin inhibitor 8 exhibits potent anti-proliferative activity against a wide range of cancer cell lines. It inhibits tubulin polymerization with an IC₅₀ of 0.73 μM. The compound suppresses the growth of K562 leukemia cells with an IC₅₀ of 14 nM. It demonstrates good anti-proliferative effectiveness against NCI-H460 (lung), SKOV3 (ovarian), BT549 (breast), 451LU (breast), SW480 (colon), COLO-205 (colon), and DLD-1 (colon) tumor cell lines, with IC₅₀ values of 15, 6, 8, 2, 8, 6, and 1 nM, respectively. These low nanomolar IC₅₀ values indicate that the compound is highly potent against a variety of cancer cell types. The compound prevents mitosis by arresting cells in the G2/M phase. This cell cycle arrest is a characteristic effect of tubulin inhibitors. The compound's potent antiproliferative activity makes it a promising candidate for further development as an anticancer agent. |
| ln Vivo |
Detailed in vivo activity data for Tubulin inhibitor 8 is not extensively reported in the available literature. However, its potent in vitro antiproliferative activity against a variety of cancer cell lines suggests that it may have potential as an anticancer agent in vivo. Typically, the in vivo efficacy of tubulin inhibitors is evaluated in xenograft mouse models. In such a study, immunodeficient mice are implanted with cancer cells, such as K562 or DLD-1 cells. When tumors reach a certain size, Tubulin inhibitor 8 is administered orally or intraperitoneally at various doses. Tumor volume is measured regularly, and final tumor weights are recorded. The compound's ability to inhibit tumor growth is assessed by comparing tumor volumes and weights between treated and control groups. The compound's in vivo activity would depend on its pharmacokinetic properties, such as oral bioavailability and tissue distribution. The compound's potent in vitro activity suggests that it could be effective in vivo at relatively low doses. Further studies are needed to fully evaluate the compound's in vivo anticancer activity.
|
| Enzyme Assay |
The inhibition of tubulin polymerization by Tubulin inhibitor 8 is typically measured using a spectrophotometric assay. The assay is performed in a 96-well plate format. Tubulin is purified from bovine brain or obtained from a commercial source. The assay mixture contains 100 mM PIPES buffer (pH 6.9), 1 mM GTP, 1 mM MgCl₂, 10% glycerol, and varying concentrations of the test compound. The reaction is initiated by the addition of tubulin. The polymerization of tubulin into microtubules is monitored by measuring the increase in absorbance at 340 nm, which is proportional to the turbidity of the microtubule solution. The increase in absorbance is monitored over time using a microplate reader. The percentage of inhibition of tubulin polymerization at each compound concentration is calculated relative to a control without inhibitor. The IC₅₀, representing the concentration that inhibits polymerization by 50%, is determined from the dose-response curve. This assay provides a direct measure of the compound's ability to inhibit tubulin polymerization.
|
| Cell Assay |
For in vitro cellular assays, the antiproliferative activity of Tubulin inhibitor 8 is typically assessed using various cancer cell lines, such as K562, NCI-H460, SKOV3, BT549, 451LU, SW480, COLO-205, and DLD-1. Cells are seeded in 96-well plates at a density of approximately 5 × 10³ cells per well and allowed to attach overnight. The cells are then treated with Tubulin inhibitor 8 at various concentrations (e.g., 0.001-10 μM) for 48-72 hours. Cell viability is assessed using an MTT or resazurin assay. In the MTT assay, viable cells with active mitochondria reduce MTT to a purple formazan product, which is solubilized and measured spectrophotometrically. The absorbance is directly proportional to the number of viable cells. The percentage of cell viability at each compound concentration is calculated relative to untreated control cells. The IC₅₀, representing the concentration that reduces cell viability by 50%, is determined from the dose-response curve. Cell cycle analysis is performed by flow cytometry using propidium iodide staining to assess the proportion of cells in the G2/M phase.
|
| Animal Protocol |
For in vivo efficacy studies, a xenograft mouse model is commonly employed. Six- to eight-week-old female athymic nude mice are injected subcutaneously in the flank with 5 × 10⁶ cancer cells (e.g., K562 or DLD-1 cells) suspended in Matrigel. When tumors reach a volume of approximately 100-150 mm³, the mice are randomized into treatment groups (n=6-8 per group). Tubulin inhibitor 8 is formulated in a suitable vehicle, such as a solution in DMSO/PEG300/Tween-80/saline. The compound is administered orally or intraperitoneally at various doses (e.g., 1, 5, 10 mg/kg) once daily or every other day for a period of 2-3 weeks. A control group receives the vehicle alone. Tumor volume is measured every 2-3 days using calipers, and body weight is recorded. At the end of the study, the mice are euthanized, and tumors are excised and weighed. The efficacy of the compound is evaluated by comparing tumor growth curves and final tumor weights between treated and control groups. Tumor tissues may be collected for histological analysis and biomarker studies to assess the compound's effects on tubulin polymerization and cell cycle progression.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties for Tubulin inhibitor 8 are not extensively reported in the available literature. The compound has a molecular weight of 342.35 and a logP of 3.8. These properties suggest that the compound is moderately lipophilic and may have reasonable oral bioavailability. The compound is soluble in DMSO. However, specific pharmacokinetic parameters such as half-life, volume of distribution, clearance, and oral bioavailability have not been determined experimentally. The compound's metabolism is likely to involve hepatic cytochrome P450 enzymes. The compound's elimination route is unknown. Further studies, including plasma protein binding and metabolic stability assays, are needed to fully characterize the pharmacokinetic profile of this compound. The compound's in vivo activity would depend on its ability to reach target tissues in sufficient concentrations.
|
| Toxicity/Toxicokinetics |
Toxicological data for Tubulin inhibitor 8 is limited, as it is a research chemical. Standard safety precautions should be observed when handling this compound. No specific toxicity studies, such as acute or chronic toxicity in animal models, have been detailed in the public domain. As a tubulin inhibitor, the compound is expected to have antiproliferative effects on rapidly dividing cells, which could lead to side effects such as myelosuppression and gastrointestinal toxicity. However, these have not been assessed. The compound is not approved for clinical use and should only be used in preclinical research settings.
|
| References |
[1]. Prinz H, et al. N-benzoylated phenoxazines and phenothiazines: synthesis, antiproliferative activity, and inhibition of tubulin polymerization. J Med Chem. 2011 Jun 23;54(12):4247-63.
|
| Additional Infomation |
Tubulin inhibitor 8 is a research tool for studying the role of microtubules in cell division and for developing new anticancer therapies. Its mechanism of action involves inhibition of tubulin polymerization. It has an IC₅₀ of 0.73 μM for tubulin polymerization. It is not approved for clinical use.
|
| Molecular Formula |
C21H14N2O3
|
|---|---|
| Molecular Weight |
342.35
|
| Exact Mass |
342.1
|
| CAS # |
1309925-39-0
|
| PubChem CID |
53262871
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
3.8
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
26
|
| Complexity |
563
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
COC1=CC=C(C=C1)C(=O)N2C3=C(C=C(C=C3)C#N)OC4=CC=CC=C42
|
| InChi Key |
JNDAHXOWDRYLBK-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C21H14N2O3/c1-25-16-9-7-15(8-10-16)21(24)23-17-4-2-3-5-19(17)26-20-12-14(13-22)6-11-18(20)23/h2-12H,1H3
|
| Chemical Name |
10-(4-methoxybenzoyl)phenoxazine-3-carbonitrile
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 2.9210 mL | 14.6049 mL | 29.2099 mL | |
| 5 mM | 0.5842 mL | 2.9210 mL | 5.8420 mL | |
| 10 mM | 0.2921 mL | 1.4605 mL | 2.9210 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
