Anticancer agent 109 (compound 6-15) is an inhibitor (blocker/antagonist) of the Gas6-Axl axis and has tumor suppressor activity. Anticancer agent 109 inhibits the expression of Gas6 and Axl, inhibits the expression of p-PI3K and p-AKT, leads to arrest in the G1 phase of the tumor cell cycle, promotes cell apoptosis, and significantly inhibits tumor growth in nude mouse tumor-bearing models.

Physicochemical Properties

Molecular Formula	C19H15N3O2
Molecular Weight	317.34
CAS #	2097497-16-8
PubChem CID	129116328
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	1
Heavy Atom Count	24
Complexity	535
Defined Atom Stereocenter Count	0
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	Axl
ln Vitro	Anticancer agent 109 (10 μM, 48 hours) induces G1 phase arrest and promotes late apoptosis, but does not alter DNA synthesis. It inhibits Gas6 and Axl in A549 cells and inhibits proteins related to the Gas6-Axl axis in PANC-1 cells [1].
ln Vivo	Anticancer agent 109 (3 mg/kg, i.p., 1 day for A549 tumor model 3, 6 times a week for 85 days for PANC-1 tumor model) significantly reduced tumor size and weight in nude mouse xenograft models[1].
Cell Assay	Cell Viability Assay[1] Cell Types: MCF-7; PANC-1; MDA-MB-231; HT-29; DU145; U937; A549; PANC-1 Tested Concentrations: 30 μM Incubation Duration: 48 h Experimental Results: Inhibited the growth of cancer cells, and was up to 20-fold safer against normal cells and up to 5.4-fold more active than Sunitinib against the cancer cells. Inhibited growth with IC50s of 2.0 μM (MCF-7); 2.8 μM (MDA-MB- 231); 4.6 μM (HT-29); 1.1 μM (DU145); 6.7 μM (U937); 4.2 μM (A549); 4.0 μM (PANC-1). Apoptosis Analysis[1] Cell Types: PANC-1 Tested Concentrations: 1 μM , 5 μM , 10 μM Incubation Duration: 48 h Experimental Results: Increased the sub-G1 fraction and induced late apoptosis. Western Blot Analysis[1] Cell Types: A549; PANC-1 Tested Concentrations: 10 μM Incubation Duration: 48 h Experimental Results: Inhibited Gas6 and Axl in A549 and PANC-1 cell, increased the expression ratio of Bax/Bcl-2 and inhibited p-PI3K and p-AKT in PANC-1 cell. RT-PCR[1] Cell Types: PANC-1 Tested Concentrations: 3 μM; 5 μM 10 μM Incubation Duration: 48 h Experimental Results: Inhibited Gas6 and Axl.
Animal Protocol	Animal/Disease Models: A549 or PANC-1 xenografted in BALB/c-nu (nude) mice [1]. Doses: 1 mg/kg; 3 mg/kg Route of Administration: intraperitoneal (ip) injection 6 times a week Experimental Results: Promoted tumor regression to around a quarter with 1 mg/kg, smaller but not eliminated with 3 mg/kg in A549 models. Promoted tumor regression to around a quarter with 3 mg/kg in PANC-1 models.
References	[1]. Antitumor effect of 3-(quinolin-2-ylmethylene)-4,6-dimethyl-5-hydroxy-7-azaoxindole down-regulating the Gas6-Axl axis. Eur J Med Chem. 2023 May 5;251:115274.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.1512 mL	15.7560 mL	31.5119 mL
	5 mM	0.6302 mL	3.1512 mL	6.3024 mL
	10 mM	0.3151 mL	1.5756 mL	3.1512 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.