ML141 (also called CID-2950007; ML-141; CID2950007; ML 141) is a novel potent, selective and reversible non-competitive inhibitor of Rho family GTPase cdc42 with potential anticancer activity. With an IC50 of 200 nM, it suppresses CDC42. By causing both cell death and division inhibition, ML141 improves TMX's capacity to inhibit the growth of BLBC cells. A substantial defense against metformin-induced apoptosis is provided by ML141 to neuroblastoma cells. In addition, ML141 reduces K. pneumoniae invasion in a manner that is dependent on dosage.

Physicochemical Properties

Molecular Formula	C22H21N3O3S
Molecular Weight	407.49
Exact Mass	407.13
Elemental Analysis	C, 64.85; H, 5.19; N, 10.31; O, 11.78; S, 7.87
CAS #	71203-35-5
Related CAS #	71203-35-5
PubChem CID	2950007
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Boiling Point	622.9±65.0 °C at 760 mmHg
Melting Point	216 °C
Flash Point	330.5±34.3 °C
Vapour Pressure	0.0±1.8 mmHg at 25°C
Index of Refraction	1.652
LogP	1.82
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	668
Defined Atom Stereocenter Count	0
SMILES	S(C1C([H])=C([H])C(=C([H])C=1[H])N1C([H])(C2C([H])=C([H])C(=C([H])C=2[H])OC([H])([H])[H])C([H])([H])C(C2C([H])=C([H])C([H])=C([H])C=2[H])=N1)(N([H])[H])(=O)=O
InChi Key	QBNZBMVRFYREHK-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H21N3O3S/c1-28-19-11-7-17(8-12-19)22-15-21(16-5-3-2-4-6-16)24-25(22)18-9-13-20(14-10-18)29(23,26)27/h2-14,22H,15H2,1H3,(H2,23,26,27)
Chemical Name	4-[3-(4-methoxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]benzenesulfonamide
Synonyms	CID-2950007;ML-141; CID2950007; ML 141;CID 2950007; ML141
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

Targets	cdc42 (IC50 = 200 nM)
ln Vitro	ML141 (CID-2950007) is not cytotoxic in either cell line at doses of 0.1-3 μM after treatment for 4 days. Ten milligrams of the compound did not cause any cytotoxicity in OVCA429 cells, but after four days of treatment, SKOV3ip cells showed some cytotoxicity at this concentration, though not to the point of statistical significance. When applied at 10 μM for 24 or 48 hours, respectively, ML141 does not cause any harm to Swiss 3T3 or Vero E6 cells[1]. ML141 suppresses the migration of ovarian cancer cells and prevents the formation of 3T3 fibroblast filopodia[1].
ln Vivo	ML141 (CID-2950007) (10 μM; intracerebroventricular injection) induces acute anxiety in mice[3].
Enzyme Assay	Overnight at 4°C, wild-type GST-Cdc42 (4 μM) is bound to GSH-beads. By incubating with a buffer containing 10 mM EDTA for 20 minutes at 30°C, washing twice with NP-HPS buffer, and then re-suspending in the same buffer containing 1 mM EDTA/or 1 mM MgCl2, 1 mM DTT, and 0.1% BSA, Cdc42 on GSH-beads is nucleotide-depleted. A 15-minute RT incubation of the protein-bead complex blocks Cdc42 unbound sites. After incubating 30 μL of this suspension with a 20 mM inhibitor for three minutes at room temperature, 30 μL of different concentrations of ice-cold BODIPY-FL-GTP are added. Samples are incubated for 45 minutes at 4° C, and an Accuri flow cytometer is used to measure the binding of fluorescent nucleotide to enzyme. The program GraphPad Prism is used to export and plot raw data.
Cell Assay	The adherent cell CeligoTM cytometer is used to count the number of live and dead cells, which are indicated by positive Calcein-AM and PI staining, respectively, after the cells are incubated with 500 nM Calcein-AM and 1 µM PI for 15 minutes.
Animal Protocol	NOD/SCID mice bearing MDA-MB 231 derived tumors 1 mg/day i.p.
References	[1]. A Potent and Selective Inhibitor of Cdc42 GTPase. [2]. EMBO Mol Med . 2013 May;5(5):723-36. [3]. Oncotarget . 2014 Nov 30;5(22):11709-22. [4]. Infect Immun . 2015 Feb;83(2):769-79. [5]. Int J Hematol . 2015 Jan;101(1):5-12.
Additional Infomation	4-[3-(4-methoxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]benzenesulfonamide is a sulfonamide.

Solubility Data

Solubility (In Vitro)

DMSO: ~81 mg/mL (~198.8 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.14 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 2: 2.08 mg/mL (5.10 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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 3: ≥ 2.08 mg/mL (5.10 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 4: 2% DMSO +30% PEG 300 +5% Tween 80 +ddH2O: 10mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4540 mL	12.2702 mL	24.5405 mL
	5 mM	0.4908 mL	2.4540 mL	4.9081 mL
	10 mM	0.2454 mL	1.2270 mL	2.4540 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.