Tubulin inhibitor 1 is a tubulin inhibitor that targets the colchicine binding site and inhibits tubulin aggregation. Tubulin inhibitor 1 has anti-tumor activity and can cause mitotic arrest in the G2/M phase of cells and induce cell apoptosis.

Physicochemical Properties

Molecular Formula	C21H24N2O4
Molecular Weight	368.426265716553
Exact Mass	368.173
CAS #	2237054-53-2
PubChem CID	137319628
Appearance	White to off-white solid powder
LogP	3.8
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	7
Heavy Atom Count	27
Complexity	430
Defined Atom Stereocenter Count	0
SMILES	O(C)C1C(=C(C=C(C=1)C1C(=CN(C)N=1)C1C=CC(=CC=1)OCC)OC)OC
InChi Key	YWYCRTNRTJCKHS-UHFFFAOYSA-N
InChi Code	InChI=1S/C21H24N2O4/c1-6-27-16-9-7-14(8-10-16)17-13-23(2)22-20(17)15-11-18(24-3)21(26-5)19(12-15)25-4/h7-13H,6H2,1-5H3
Chemical Name	4-(4-ethoxyphenyl)-1-methyl-3-(3,4,5-trimethoxyphenyl)pyrazole
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)

Biological Activity

ln Vitro	Tubulin inhibitor 1 (compound 7a3) is a tubulin inhibitor that prevents tubulin polymerization [1]. Strong anti-proliferative activity of tubulin inhibitor 1 has been observed against SK-OV-3, MDA-MB-231, HeLa, A549, CT26, and MCF-7 cells; IC50 values are 16.7 ± 3.0, 31.4 ± 0.7, 32.8 ± 2.9, 67.0 ± 0.8, 58.0 ± 2.4, and 35.4 ± 5.6 nM, in that order [1]. In SK-OV-3 cells, tubulin inhibitor 1 (40, 80, and 160 nM, 48 hours) dramatically stopped cell mitosis in the G2/M phase and triggered apoptosis [1].
ln Vivo	Tubulin Inhibitor 1 (50 mg/kg, i.p., three times every two days for 20–25 days) significantly reduced the number of SK-OV-3 cell-bearing Balb/c nude mice while also being well tolerated. of tumor development [1].
Cell Assay	Apoptosis analysis [1] Cell Types: SK-OV-3 Cell Tested Concentrations: 40, 80 and 160 nM Incubation Duration: 48 hrs (hours) Experimental Results: SK-OV-3 cells induced apoptosis after 48 hrs (hours) of treatment.
Animal Protocol	Animal/Disease Models: Sixweeks old Balb/c nude mice (18-20 g), carrying SK-OV-3 cells [1] Doses: 50 mg/kg Route of Administration: IP, 3 times every two days for 20-25 Experimental Results: The tumor growth of Balb/c nude mice carrying SK-OV-3 cells was Dramatically diminished, but the body weight was not Dramatically diminished.
References	[1]. Design, synthesis and biological evaluation of a novel tubulin inhibitor 7a3. Eur J Med Chem. 2018 Aug 5;156:162-179.

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 125 mg/mL (~339.28 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.65 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 20.8 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: ≥ 2.08 mg/mL (5.65 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.08 mg/mL (5.65 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 20.8 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	2.7142 mL	13.5711 mL	27.1422 mL
	5 mM	0.5428 mL	2.7142 mL	5.4284 mL
	10 mM	0.2714 mL	1.3571 mL	2.7142 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.