| ln Vitro |
Tubulin-IN-67 (compound 15u) (72 hours) effectively inhibited the proliferation of Huh7, HeLa and MCF-7 cancer cells, with IC50 values of 0.090 μM, 0.10 μM and 0.12 μM, respectively, and had a significantly better safety window compared with CA-4 in normal HUVEC cells [1]. Tubulin-IN-67 (0.5-1 μM) is a direct microtubule destabilizing compound that effectively inhibits the polymerization of microtubules purified in vitro [1]. Tubulin-IN-67 (90-180 nM; 24 hours) disrupted the microtubule network in Huh7 cells and induced perinuclear microtubule aggregation, which is consistent with the microtubule inhibition effect of colchicine sites [1]. Tubulin-IN-67 (90-270 nM; 24 hours) can induce strong, dose-dependent cell cycle arrest in the G2/M phase of Huh7 cells, with the highest tested concentration (270 nM) causing 83.16% of cells to arrest in the G2/M phase [1]. Tubulin-IN-67 (90-270 nM; 48 hours) can effectively induce dose-dependent apoptosis in Huh7 cells, with a total apoptosis rate of 46.6% at the highest tested concentration of 270 nM [1].
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|---|---|
| ln Vivo |
Tubulin-IN-67 (compound 15u) (15 mg/kg; intraperitoneal injection; once every other day for 14 days) significantly inhibited tumor growth in a mouse model of orthotopic hepatocellular carcinoma and had very low systemic toxicity, with efficacy comparable to paclitaxel [1].
|
| Cell Assay |
Immunofluorescence [1]
Cell Types: Huh7 human hepatocellular carcinoma cells Tested Concentrations: 90 nM; 180 nM Incubation Duration: 24 hours Experimental Results: Induced significant morphological changes in the microtubule cytoskeleton, leading to microtubule shortening and condensation around the nucleus, similar to the effect of colchicine site binder CA-4 (90 nM). Cell cycle analysis [1] Cell Types: Huh7 human liver cancer cells Tested Concentrations: 90 nM; 180 nM; 270 nM Incubation Duration: 24 hours Experimental Results: The treatment resulted in a dose-dependent accumulation of cells in the G2/M phase. The proportion of the untreated control group was 4.0%, which increased to 13.69% (90 nM), 69.46% (180 nM) and 83.16% (270 nM), respectively. Apoptosis analysis [1] Cell Types: Huh7 human liver cancer cells Tested Concentrations: 90 nM; 180 nM; 270 nM Incubation Duration: 48 hours Experimental Results: Apoptosis was induced in a concentration-dependent manner, and the total apoptosis rates (early and late stages) were 15.5% (90 nM), 27.0% (180 nM) and 46.6% (270 nM), respectively. |
| Animal Protocol |
Animal/Disease Models:BALB/c nude mice (male, 5 weeks old, SPF grade; orthotopic hepatocellular carcinoma model established by surgical implantation of Huh7 cells) [1]
Doses: 15 mg/kg Route of Administration: Intraperitoneal injection; every other day; for 14 days Experimental Results: Compared with the vector control group, it significantly inhibited tumor progression, and the efficacy was comparable to paclitaxel. It maintained stable body weight without significant decrease. The liver tissue structure was intact, with no hepatocellular damage or inflammatory infiltration, while the tumor section showed a large area of necrosis, which was significantly increased compared with the vector group. |
| References |
| Molecular Formula |
C23H20N4O3
|
|---|---|
| Molecular Weight |
400.43
|
| Appearance |
Typically exists as solids at room temperature
|
| SMILES |
COC1=C(OC)C(OC)=CC(N2N=CC3=C2C=C(C4=CC=CC5=C4C=CN5)N=C3)=C1
|
| 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
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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 | 2.4973 mL | 12.4866 mL | 24.9732 mL | |
| 5 mM | 0.4995 mL | 2.4973 mL | 4.9946 mL | |
| 10 mM | 0.2497 mL | 1.2487 mL | 2.4973 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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