Vimentin-IN-1 is a FiVe1 analogue and an orally bioactive selective anti-cancer compound. FiVe1 binds to the type III intermediate filament protein vimentin (VIM) and induces Ser56 hyperphosphorylation, leading to selective interruption of mitosis and multinucleation of mesenchymal cancer/tumor cells transformed to express VIM. Vimentin-IN-1 displays better oral availability and pharmacokinetic characteristics than FiVe1.

Physicochemical Properties

Molecular Formula	C19H18CL2N4O
Molecular Weight	389.278421878815
Exact Mass	388.085
CAS #	2319587-80-7
PubChem CID	156552418
Appearance	Light yellow to orange solid powder
LogP	4.1
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	3
Heavy Atom Count	26
Complexity	463
Defined Atom Stereocenter Count	0
InChi Key	JWIRSMDIIXNJAU-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H18Cl2N4O/c1-26-15-10-13(20)9-14(11-15)24-5-7-25(8-6-24)18-12-22-23-19-16(18)3-2-4-17(19)21/h2-4,9-12H,5-8H2,1H3
Chemical Name	8-chloro-4-[4-(3-chloro-5-methoxyphenyl)piperazin-1-yl]cinnoline
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	Vimentin-IN-1 (compound 4e) (0-10 mM; 72 h) is much more potent than FiVe1 (IC50=1.6 μM, HT-1080) at inhibiting HT-1080 fibrosarcoma, with an IC50 value of 44 nM [1]. Phosphorylation of VIM Ser56 is induced by venterin-IN-1 (0.1 μM; 24 hours) [1]. In mouse liver microsomes, ventin-IN-1 (100 μM) is less stable; after 60 minutes of incubation, 0.0% is still present [1]. Samples are taken at 0, 5, 15, 30, 45, and 60 minutes.
ln Vivo	The oral pharmacokinetic properties of Vimentin-IN-1 (Compound 4e) (10 mg/kg; single dose) are superior to those of Five1 [1]. Vimentin-IN-1's pharmacokinetic characteristics in mice [1] The dosage by route (mg/kg) AUC0-last (ng·h/mL) AUC0-inf (ng·h/mL) T1/2 (hour) Tmax (h), Cmax (ng/mL), and Final (h) 4e PO 25 309.78 339.21 4.57 0. 5 18 110.43 PO 10 371.33 534.33 4.68 0.67 8 154.67 4e IP 1 208.33 211.33 0.59 0.25 4 197.00 Five1
Cell Assay	Cell Viability Assay[1] Cell Types: HT-1080, RD and MCF-7 Cell Tested Concentrations: 0-10 mM Incubation Duration: 72 hrs (hours) Experimental Results: Inhibition of HT-1080, RD and MCF-7 cells with IC50 of 44 nM, 61 nM and 49 nM respectively.
References	[1]. Synthesis and biological evaluation of novel FiVe1 derivatives as potent and selective agents for the treatment of mesenchymal cancers. Eur J Med Chem. 2022 Nov 15;242:114638.

Solubility Data

Solubility (In Vitro)

DMSO : ~25 mg/mL (~64.22 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (6.42 mM) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.5688 mL	12.8442 mL	25.6885 mL
	5 mM	0.5138 mL	2.5688 mL	5.1377 mL
	10 mM	0.2569 mL	1.2844 mL	2.5688 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.