L-006235 (L-235) is a potent, selective, reversible and orally bioactive cathepsin K inhibitor (antagonist) with IC50 of 5 nM in a bone resorption assay. L-006235 has selectivity for cathepsin K (Ki=0.2 nM) over cathepsin B, cathepsin L and cathepsin S (Ki=1, 6, 47 μM). L-006235 reduces collagen degradation and prevents bone loss.

Physicochemical Properties

Molecular Formula	C24H30N6O2S
Molecular Weight	466.6
Exact Mass	466.215
CAS #	294623-49-7
PubChem CID	9912381
Appearance	White to off-white solid powder
LogP	3.868
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	33
Complexity	733
Defined Atom Stereocenter Count	0
InChi Key	FIVYCSWOCXEWSE-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H30N6O2S/c1-29-13-15-30(16-14-29)23-27-20(17-33-23)18-5-7-19(8-6-18)21(31)28-24(9-3-2-4-10-24)22(32)26-12-11-25/h5-8,17H,2-4,9-10,12-16H2,1H3,(H,26,32)(H,28,31)
Chemical Name	N-[1-(cyanomethylcarbamoyl)cyclohexyl]-4-[2-(4-methylpiperazin-1-yl)-1,3-thiazol-4-yl]benzamide
Synonyms	L-006235; L 006235; L006235
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	In the rabbit bone resorption experiment, L-006235 inhibits bone resorption with an IC50 of 5 nM[1]. In HepG2 cells, L-006235 (10 μM; 1 h) exhibits punctate fluorescence throughout the cytoplasm [2].
ln Vivo	L-006235, a suggested share nutrient, decreases N-telopeptide (NTx) and creatine nutrient (Cre) in rhesus monkeys by up to 76% when administered poqd for 8-11 days [1]. L-006235 (20 mg/kg; po; deposition) has a lengthy terminal half-life (204 minutes), a high bioavailability (68%), and a Cmax (1.4 μM) [1].
Animal Protocol	Animal/Disease Models: Rhesus monkey (15 years old) underwent ovariectomy (OVX) [1] Doses: 0.6, 3, 15 mg/kg Route of Administration: Orally one time/day for 8-11 days Experimental Results: Average reduction in uNTx/Cre The 76%, 68% and 31% doses were 15, 3 and 0.6mg/kg respectively.
References	[1]. Design and synthesis of tri-ring P3 benzamide-containing aminonitriles as potent, selective, orally effective inhibitors of cathepsin K. J Med Chem. 2005 Dec 1;48(24):7520-34. [2]. Lysosomotropism of basic cathepsin K inhibitors contributes to increased cellular potencies against off-target cathepsins and reduced functional selectivity. J Med Chem. 2005 Dec 1;48(24):7535-43. [3]. Cathepsin K inhibitors prevent bone loss in estrogen-deficient rabbits. J Bone Miner Res. 2011 Feb;26(2):252-62.

Solubility Data

Solubility (In Vitro)

DMSO : ~31.25 mg/mL (~66.97 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.46 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 (4.46 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 (4.46 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	2.1432 mL	10.7158 mL	21.4316 mL
	5 mM	0.4286 mL	2.1432 mL	4.2863 mL
	10 mM	0.2143 mL	1.0716 mL	2.1432 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.