KLH45 is a potent and specific DDHD2 inhibitor (antagonist) with IC50 of 1.3 nM.

Physicochemical Properties

Molecular Formula	C24H25F3N4O2
Molecular Weight	458.476115942001
Exact Mass	458.192
Elemental Analysis	C, 62.87; H, 5.50; F, 12.43; N, 12.22; O, 6.98
CAS #	1632236-44-2
PubChem CID	126970670
Appearance	Off-white to light yellow solid powder
Density	1.3±0.1 g/cm3
Boiling Point	561.0±60.0 °C at 760 mmHg
Flash Point	293.1±32.9 °C
Vapour Pressure	0.0±1.5 mmHg at 25°C
Index of Refraction	1.591
LogP	5.87
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	33
Complexity	615
Defined Atom Stereocenter Count	0
SMILES	N1=CC(C2=CC=C(OC(F)(F)F)C=C2)=NN1C(N(C1CCCCC1)CCC1=CC=CC=C1)=O
InChi Key	FYOYNRLSBYWAHL-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H25F3N4O2/c25-24(26,27)33-21-13-11-19(12-14-21)22-17-28-31(29-22)23(32)30(20-9-5-2-6-10-20)16-15-18-7-3-1-4-8-18/h1,3-4,7-8,11-14,17,20H,2,5-6,9-10,15-16H2
Chemical Name	N-cyclohexyl-N-(2-phenylethyl)-4-[4-(trifluoromethoxy)phenyl]triazole-2-carboxamide
Synonyms	KLH45; KLH-45; KLH 45;
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	KLH45 (25 nM, 4 hr) has no cross-reactivity with any of the over 40 identified serine hydrolases, with the exception of ABHD6, and completely inactivates DDHD2 in Neuro2A cells (>95% inhibition)[1]. In DDHD2-expressing cells supplemented with fatty acids, KLH45 (2 μM) enhances LD formation [2].
ln Vivo	KLH45 (20 mg/kg; given every 12 hours) caused notable increases in a number of TAGs that accumulated over the course of four days in the brains of DDHD2−/− mice [1].
Cell Assay	Cell viability assay [2] Cell Types: COS-7 cells. Tested Concentrations: 2μM. Incubation Duration: 16 hrs (hours). Experimental Results: Selective blocking of DDHD2 activity.
Animal Protocol	Animal/Disease Models: Mouse[1]. Doses: 5-40 mg/kg. Management: IP, once. Experimental Results: No altered brain signatures were shown.
References	[1]. The hereditary spastic paraplegia-related enzyme DDHD2 is a principal brain triglyceride lipase. Proc Natl Acad Sci U S A. 2014 Oct 14;111(41):14924-9. [2]. Functional Contribution of the Spastic Paraplegia-Related Triglyceride Hydrolase DDHD2 to the Formation and Content of Lipid Droplets. Biochemistry. 2018 Feb 6;57(5):827-838.

Solubility Data

Solubility (In Vitro)

DMSO : ~180 mg/mL (~392.60 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 4.5 mg/mL (9.82 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 45.0 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: ≥ 4.5 mg/mL (9.82 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 45.0 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. Solubility in Formulation 3: ≥ 4.5 mg/mL (9.82 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 45.0 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	2.1811 mL	10.9056 mL	21.8112 mL
	5 mM	0.4362 mL	2.1811 mL	4.3622 mL
	10 mM	0.2181 mL	1.0906 mL	2.1811 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.