ZX-29 is a potent and specific ALK inhibitor (antagonist) with IC50s of 2.1 nM, 1.3 nM and 3.9 nM for ALK, ALK L1196M and ALK G1202R respectively, but has no activity against EGFR. ZX-29 causes apoptosis by inducing endoplasmic reticulum stress and overcomes cellular resistance caused by ALK mutations. ZX-29 can also induce protective autophagy and have anti-tumor effects.

Physicochemical Properties

Molecular Formula	C23H28CLN7O3S
Molecular Weight	518.03
Exact Mass	517.17
Elemental Analysis	C, 53.33; H, 5.45; Cl, 6.84; N, 18.93; O, 9.27; S, 6.19
CAS #	2254805-62-2
Related CAS #	2254805-62-2
PubChem CID	142497299
Appearance	Light yellow to green yellow solid powder
LogP	3.5
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	10
Rotatable Bond Count	8
Heavy Atom Count	35
Complexity	765
Defined Atom Stereocenter Count	0
InChi Key	ATHZZWBANOJUOA-UHFFFAOYSA-N
InChi Code	InChI=1S/C23H28ClN7O3S/c1-30-10-12-31(13-11-30)16-8-9-20(21(14-16)34-2)27-23-25-15-17(24)22(28-23)26-18-6-4-5-7-19(18)29-35(3,32)33/h4-9,14-15,29H,10-13H2,1-3H3,(H2,25,26,27,28)
Chemical Name	N-[2-[[5-chloro-2-[2-methoxy-4-(4-methylpiperazin-1-yl)anilino]pyrimidin-4-yl]amino]phenyl]methanesulfonamide
Synonyms	ZX29; ZX 29; ZX-29
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: This product requires protection from light (avoid light exposure) during transportation and storage.
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	ALK (IC50 = 2.1 nM); ALK L1196M (IC50 = 1.3 nM); ALK G1202R (IC50 = 3.9 nM)
ln Vitro	ZX-29 (0-81 nM; 24-72 hours; NCI-H2228 cells) cell viability decreases in a dose- and time-dependent manner following ZX-29 treatment[1]. ZX-29 (10 nM; 24 hours; NCI-H2228 cells) results in the production of autophagosomes and the usual symptoms of autophagy. ZX-29 raises LC3 and Beclin1 expression levels[1]. ZX-29 (10 nM; 0-48 hours; NCI-H2228 cells) inhibits NCI-H2228 cells in the G1 phase and prevents them from proliferating[1]. ZX-29 (10-40 nM; 24-48 hours; NCI-H2228 cells) cells undergo apoptosis in response to ZX-29 treatment. ZX-29 dose-dependently increases the production of activated forms of caspase 3, downregulates the expression of the antiapoptotic protein Bcl-2, and upregulates the expression levels of the proapoptotic protein Bax[1]. ZX-29 (30-300 nM; 24 hours; NCI-H2228 cells) treatment dramatically and dose-dependently reduces the expression of p-ALK and its downstream signaling proteins, such as p-Akt and p-STAT3[1]. ZX-29 (20 nM; 0-48 hours; NCI-H2228 cells) treatment significantly increases the mRNA level of CHOP[1]. ZX-29 inhibits NCI-H2228 cell colony formation in a dose-dependent manner. ZX-29 concentration increased was accompanied by a progressive decrease in cell density as well as a sharpening and slendering of the cells' normal morphology[1].
ln Vivo	ZX-29 (50 mg/kg; intragastric administration; every 2 days; for a total of 7 times; female BALB/c nude mice) treatment inhibits tumor growth in a mouse xenograft model[1].
Animal Protocol	Female BALB/c nude mice (4-week-old) with H2228 cells[1] 50 mg/kg Intragastric administration; every 2 days; for a total of 7 times
References	[1]. ZX-29, a novel ALK inhibitor, induces apoptosis via ER stress in ALK rearrangement NSCLC cells and overcomes cell resistance caused by an ALK mutation. Biochim Biophys Acta Mol Cell Res. 2020 Mar 26;1867(7):118712.

Solubility Data

Solubility (In Vitro)

DMSO: ~50 mg/mL (~96.5 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.83 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9304 mL	9.6520 mL	19.3039 mL
	5 mM	0.3861 mL	1.9304 mL	3.8608 mL
	10 mM	0.1930 mL	0.9652 mL	1.9304 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.