RU-SKI 43 HCl is a potent and specific inhibitor of hedgehog acyltransferase (Hhat) with IC50 of 850 nM. RU-SKI 43 HCl reduces Gli-1 activation through smoothing-independent non-canonical signaling and inhibits Akt and mTOR pathway activity. RU-SKI 43 HCl has anti-cancer activity.

Physicochemical Properties

Molecular Formula	C22H31CLN2O2S
Molecular Weight	423.01
Exact Mass	422.179
CAS #	1782573-67-4
Related CAS #	RU-SKI 43;1043797-53-0
PubChem CID	90488982
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	8
Heavy Atom Count	28
Complexity	475
Defined Atom Stereocenter Count	0
InChi Key	JBBKLHJLHRGJSQ-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H30N2O2S.ClH/c1-4-16(2)13-23-14-22(25)24-10-8-21-19(9-11-27-21)20(24)15-26-18-7-5-6-17(3)12-18;/h5-7,9,11-12,16,20,23H,4,8,10,13-15H2,1-3H3;1H
Chemical Name	2-(2-methylbutylamino)-1-[4-[(3-methylphenoxy)methyl]-6,7-dihydro-4H-thieno[3,2-c]pyridin-5-yl]ethanone;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

Targets	IC50: 850 nM (Hhat)[1]
ln Vitro	RU-SKI 43 hydrochloride (10 μM; for 6 days) greatly inhibits cell proliferation (83% in AsPC-1 cells) in AsPC-1 and Panc-1 cells[2]. RU-SKI 43 hydrochloride (10 or 20 μM; 5 hours) produces dose-dependent suppression of Shh palmitoylation following only 5 hours[1]. RU-SKI 43 hydrochloride (10 μM; for 72 hours) causes a 40% drop in Gli-1 levels in AsPC-1 cells[2] . RU-SKI 43 hydrochloride (10 μM; 48 hours) leads in reduced phosphorylation (47-67%) of four proteins in the Akt pathway, including Akt (phosphorylation at both Thr307 and Ser473), PRAS40, Bad and GSK-3β. RU-SKI 43 therapy also reduces phosphorylation of mTOR and S6, components of the mTOR signaling pathway[2]. RU-SKI 43 hydrochloride functions as an uncompetitive inhibitor (Ki=7.4 μM) with regard to Shh, and as a noncompetitive inhibitor ( Ki=6.9 μM) with respect to 125I-iodo-palmitoylCoA[1].
ln Vivo	After being administered intravenously, RU-SKI 43 hydrochloride has a t1/2 of 17 min in mouse plasma[1].
Cell Assay	Cell Proliferation Assay[2] Cell Types: AsPC-1 and Panc-1 pancreatic cancer cells Tested Concentrations: 10 μM Incubation Duration:For 6 days (drugs were replenished every 48 hrs (hours)) Experimental Results: Strongly diminished cell proliferation (83% in AsPC-1 cells). Western Blot Analysis[1] Cell Types: COS-1 cells expressing HA-Hhat and Shh Tested Concentrations: 10 or 20 μM Incubation Duration: 5 hrs (hours) Experimental Results: Caused dose-dependent inhibition of Shh palmitoylation following only 5 hrs (hours).
References	[1]. Inhibitors of Hedgehog acyltransferase block Sonic Hedgehog signaling.Nat Chem Biol. 2013 Apr;9(4):247-9. [2]. Hedgehog acyltransferase as a target in pancreatic ductal adenocarcinoma. Oncogene. 2014 Jan 27. doi: 10.1038/onc.2013.575.

Solubility Data

Solubility (In Vitro)

DMSO: ≥ 51 mg/mL (120.56 mM) H2O: 2.5 mg/mL (5.91 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.91 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 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.91 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 25.0 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.3640 mL	11.8201 mL	23.6401 mL
	5 mM	0.4728 mL	2.3640 mL	4.7280 mL
	10 mM	0.2364 mL	1.1820 mL	2.3640 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.