Werner syndrome RecQ helicase-IN-3 is a potent orally active Werner syndrome RecQ DNA helicase WRN inhibitor with an IC50 value of 0.06 μM. RecQ helicase-IN-3 with Werner syndrome has antiproliferative properties. RecQ helicase-IN-3 from Werner syndrome exhibits anticancer activity.

Physicochemical Properties

Molecular Formula	C31H30CLF3N8O5
Molecular Weight	687.068715572357
Exact Mass	686.2
CAS #	2869954-98-1
PubChem CID	166140846
Appearance	White to off-white solid powder
LogP	4.2
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	13
Rotatable Bond Count	7
Heavy Atom Count	48
Complexity	1290
Defined Atom Stereocenter Count	0
SMILES	C12=NC(C3=CCOCC3)=NN1C(=O)C(N1CCN(C(C3=NC=CC=C3O)=O)CC1)=C(CC)N2CC(NC1=CC=C(C(F)(F)F)C=C1Cl)=O
InChi Key	JXPFVKAENGSGMB-UHFFFAOYSA-N
InChi Code	InChI=1S/C31H30ClF3N8O5/c1-2-22-26(40-10-12-41(13-11-40)28(46)25-23(44)4-3-9-36-25)29(47)43-30(38-27(39-43)18-7-14-48-15-8-18)42(22)17-24(45)37-21-6-5-19(16-20(21)32)31(33,34)35/h3-7,9,16,44H,2,8,10-15,17H2,1H3,(H,37,45)
Chemical Name	-[2-chloro-4-(trifluoromethyl)phenyl]-2-[2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-[4-(3-hydroxypyridine-2-carbonyl)piperazin-1-yl]-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]acetamide
Synonyms	Werner syndrome RecQ helicase-IN-3
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	WRN ( IC50 = 0.06 µM )
ln Vitro	RecQ helicase-IN-3 (example 96) for Werner syndrome exhibits anti-proliferative action at GI50s of >10 µM for DLD1 WRN-KO cells and 0.06 for SW48 cells, respectively, during 8–20 days [1].
ln Vivo	Mice treated daily for 22 days with 120 mg/kg of Werner syndrome RecQ helicase-IN-3 exhibit antitumor action [1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: SW48, DLD1 WRN-KO cells Tested Concentrations: 0-10 µM Incubation Duration: 8-20 days Experimental Results: demonstrated antiproliferative activity with GI50s of 0.06, >10 µM for SW48, DLD1 WRN-KO cells, respectively.
Animal Protocol	Animal/Disease Models: Female Crl:NU(NCr)-Foxn1nu -Homozygous nude mice (SW48 xenografts cells)[1] Doses: 120 mg/kg Route of Administration: Po ; daily for 22 days Experimental Results: Inhibited tumor growth.
References	[1]. Triazolo-pyrimidine analogues for treating diseases connected to the inhibiton of werner syndrome recq helicase (wrn). WO2022249060A1.

Solubility Data

Solubility (In Vitro)

DMSO : ~62.5 mg/mL (~91 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.03 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 (3.03 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 (3.03 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	1.4555 mL	7.2773 mL	14.5546 mL
	5 mM	0.2911 mL	1.4555 mL	2.9109 mL
	10 mM	0.1455 mL	0.7277 mL	1.4555 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.