ML-9 HCl is a novel, potent and selective myosin light chain kinase (MLCK) inhibitor, also Akt kinase and and stromal interaction molecule 1 (STIM1). ML-9 enhances the anticancer activity of docetaxel, suggesting its potential application as an adjuvant to existing anticancer chemotherapy. By simultaneously inhibiting the Akt pathway and autophagy, ML-9 was found to have a complex effect on autophagy and to be a useful tool for targeting autophagy in cancer therapy.

Physicochemical Properties

Molecular Formula	C15H18CL2N2O2S
Molecular Weight	361.28662
Exact Mass	360.046
Elemental Analysis	C, 49.87; H, 5.02; Cl, 19.63; N, 7.75; O, 8.86; S, 8.88
CAS #	105637-50-1
Related CAS #	ML-9 Free Base;110448-31-2
PubChem CID	108047
Appearance	Off-white to light yellow solid powder
Boiling Point	508.8ºC at 760mmHg
Melting Point	196-200ºC
Flash Point	261.5ºC
Vapour Pressure	1.79E-10mmHg at 25°C
LogP	4.626
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	22
Complexity	451
Defined Atom Stereocenter Count	0
SMILES	C1=CC2=C(C=CC=C2S(=O)(=O)N3CCCNCC3)C(=C1)Cl.Cl
InChi Key	ZNRYCIVTNLZOGI-UHFFFAOYSA-N
InChi Code	InChI=1S/C15H17ClN2O2S.ClH/c16-14-6-1-5-13-12(14)4-2-7-15(13)21(19,20)18-10-3-8-17-9-11-18/h1-2,4-7,17H,3,8-11H21H
Chemical Name	1-(5-chloronaphthalen-1-yl)sulfonyl-1,4-diazepane;hydrochloride
Synonyms	ML9 HCl; ML-9 HCl; ML 9 HCl; ML-9 hydrochloride
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	While ML9 (0-100 μM; 0-24 hours) does not dramatically trigger cell death, it does significantly diminish the viability of cardiac cells [2]. ML9 (50 μM; 1-4 hours) dramatically raises the amounts of mitochondrial caspase-3, allowing
Cell Assay	cell viability assay [1] Cell Types: Neonatal rat ventricular myocytes (NRVM) Cell Tested Concentrations: 0, 10, 50 and 100 μM Incubation Duration: 0, 1, 4, 8 and 24 STIM1 protein levels diminished by approximately 42% [2]. Hourly Experimental Results: diminished cell viability at concentrations of 50-100 μM. Apoptosis analysis[1] Cell Types: Neonatal rat ventricular myocytes (NRVM) Cell Tested Concentrations: 50 μM Incubation Duration: 1, 4 and 8 hrs (hours) Experimental Results: Induction of cardiomyocyte death via necrosis and apoptosis.
References	[1]. ML-9, a myosin light chain kinase inhibitor, reduces intracellular Ca2+ concentration in guinea pig trachealis.Eur J Pharmacol. 2004 Feb 23;486(3):325-33. [2]. The STIM1 inhibitor ML9 disrupts basal autophagy in cardiomyocytes by decreasing lysosome content.Toxicol In Vitro. 2018 Apr;48:121-127. [3]. Identification of ML-9 as a lysosomotropic agent targeting autophagy and cell death.Cell Death Dis. 2014 Apr 24;5:e1193.

Solubility Data

Solubility (In Vitro)

DMSO : ~83.33 mg/mL (~230.65 mM) H2O : ≥ 5 mg/mL (~13.84 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.76 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 (5.76 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 (5.76 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	2.7679 mL	13.8393 mL	27.6786 mL
	5 mM	0.5536 mL	2.7679 mL	5.5357 mL
	10 mM	0.2768 mL	1.3839 mL	2.7679 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.