JAK3-IN-1 is a potent, selective and orally bioactive JAK3 inhibitor (antagonist) with IC50 of 4.8 nM. JAK3-IN-1 is more than 180-fold more selective for JAK3 compared to JAK1 (IC50 of 896 nM) and JAK 2 (IC50 of 1050 nM).

Physicochemical Properties

Molecular Formula	C26H30CLN7O2
Molecular Weight	508.02
Exact Mass	507.214
CAS #	1805787-93-2
PubChem CID	92042864
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Index of Refraction	1.672
LogP	2.18
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	9
Heavy Atom Count	36
Complexity	708
Defined Atom Stereocenter Count	0
InChi Key	UGXCBYVBIJACEK-UHFFFAOYSA-N
InChi Code	InChI=1S/C26H30ClN7O2/c1-4-24(35)30-19-7-5-6-18(14-19)16-28-25-21(27)17-29-26(32-25)31-22-9-8-20(15-23(22)36-3)34-12-10-33(2)11-13-34/h4-9,14-15,17H,1,10-13,16H2,2-3H3,(H,30,35)(H2,28,29,31,32)
Chemical Name	N-[3-[[[5-chloro-2-[2-methoxy-4-(4-methylpiperazin-1-yl)anilino]pyrimidin-4-yl]amino]methyl]phenyl]prop-2-enamide
Synonyms	JAK3 IN 1 JAK3IN1 JAK3-IN-1
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	Treatment with JAK3-IN-1 (Compound 9; 0–5 µM; 3 hours; BMDMs cells) totally suppressed p-STAT6 elicited by IL-4 at 500 nM, but only slightly inhibited p-STAT6 induced by IFNβ at 500 nM. p-STAT1 at 5.0μM [1]. ..Several tyrosine-protein kinases (TEC) family kinases and fms-related tyrosine kinase 3 (FLT3) were shown to be possible off-targets of JAK3-IN-1 (compound 9), which most potently inhibited JAK3. The protein enzymatically inhibits the kinase TXK (TXK, IC50 = 36 nM). Enzyme assays employing Z'-lyte or LanthaScreen formats reveal detection of FLT3 (IC50 = 13 nM), TTK protein kinase (TTK, IC50 = 49 nM), BLK proto-oncogene (BLK, IC50 = 157 nM), and tyrosine. JAK3-IN-1 exhibited extremely low fractional inhibition against wild-type (WT) EGFR and other JAKs, which is in line with an IC50 against EGFRWT and TYK2 that is >180 times higher (IC50 = 409 nM and >10000, respectively). JAK3-IN-1 has an IC50 that is more than 165 times greater than that of BTK or ITK (IC50 = 794 and 1070 nM, respectively) [1]. At dosages below 3.0 μM, there is no antiproliferative impact. JAK3-IN-1(Compound 9) specifically inhibits the proliferation of JAK3-dependent Ba/F3 cells (IC50 = 69 nM) contrasted to other JAK-dependent Ba/F3 cells [1].
ln Vivo	JAK3-IN-1 (compound 9) demonstrated reasonable pharmacokinetic features, with a T1/2 of 1.4 h and an area under the curve (AUC) value of 795 ng*hr/mL after an oral dosage of 10 mg/Kg. Utilization is good at 66%. After oral administration of JAK3-IN-1 (Compound 9) (75 mpk, QD) for 8 days, the number of B or T lymphocytes in the lungs and spleens of tumor-bearing mice was not altered, while NK cells were reduced [1].
Cell Assay	Western Blot Analysis[1] Cell Types: BMDMs Cell Tested Concentrations: 0 µM, 0.1 µM, 0.5 µM, 1 µM, 5 µM Incubation Duration: 3 hrs (hours) Experimental Results: Complete inhibition of IL-4-induced p-STAT6 concentration at 500 nM At 5.0 μM, it only partially inhibits IFNβ-induced p-STAT1.
References	[1]. Development of Selective Covalent Janus Kinase 3 Inhibitors. J Med Chem. 2015 Aug 27;58(16):6589-6606.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~196.84 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.92 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 (4.92 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (4.92 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 25.0 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	1.9684 mL	9.8421 mL	19.6843 mL
	5 mM	0.3937 mL	1.9684 mL	3.9369 mL
	10 mM	0.1968 mL	0.9842 mL	1.9684 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.