AXL-IN-13 is a potent, orally bioactive AXL inhibitor (IC50= 1.6 nM, Kd: 0.26 nM). AXL-IN-13 reverses TGF-β1-induced epithelial-mesenchymal transition (EMT) and inhibits cancer cell migration and invasion.

Physicochemical Properties

Molecular Formula	C34H41FN6O5
Molecular Weight	632.724951505661
Exact Mass	632.312
CAS #	2376928-82-2
PubChem CID	139533572
Appearance	Off-white to light yellow solid powder
LogP	5.6
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	10
Rotatable Bond Count	12
Heavy Atom Count	46
Complexity	942
Defined Atom Stereocenter Count	0
SMILES	N1(C)C=C(C(NC2CCCCC2)=O)C(NC2=CC=C(OC3C4C(N=CC=3)=CC(OCCCN3CCOCC3)=C(OC)C=4)C(F)=C2)=N1
InChi Key	QBRQTJMZEYORSP-UHFFFAOYSA-N
InChi Code	InChI=1S/C34H41FN6O5/c1-40-22-26(34(42)38-23-7-4-3-5-8-23)33(39-40)37-24-9-10-30(27(35)19-24)46-29-11-12-36-28-21-32(31(43-2)20-25(28)29)45-16-6-13-41-14-17-44-18-15-41/h9-12,19-23H,3-8,13-18H2,1-2H3,(H,37,39)(H,38,42)
Chemical Name	N-cyclohexyl-3-[3-fluoro-4-[6-methoxy-7-(3-morpholin-4-ylpropoxy)quinolin-4-yl]oxyanilino]-1-methylpyrazole-4-carboxamide
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	PDGFRβ 2.3 nM (Kd)
ln Vitro	According to ELISA data, AXL-IN-13 (Compound 6li) inhibits Ba/F3-TEL-AXL cell proliferation with an IC50 of 4.7 nM [1]. Inhibition of CSF1R, FLT1/3/4, KLT, PDGFRB, and TIE2 binding affinity has also been demonstrated by AXL-IN-13 [1]. In MDA-MB-231 and 4T1 cells, AXL-IN-13 (0-500 nM, 6 hours) inhibits AXL phosphorylation [1]. In MDA-MB-231 cells, TGF-β1 (10 ng/mL)-induced EMT is blocked by AXL-IN-13 (0-3 μM, 3 days)[1]. MDA-MB-231 cells' migration and invasion caused by TGF-β1 (10 ng/mL) are inhibited by AXL-IN-13 (0-3 μM, 24 hours) [1].
ln Vivo	AXL-IN-13 (compound 6li) (50 or 100 mg/kg, oral, 14 days) inhibits 4T1 tumor growth and metastasis[1]. AXL-IN-13 (25 mg/kg, oral) shows a good PK curve, with an AUC of 8410.21 ng/mL/h, a T1/2 value of 4.22 h, and an oral bioavailability (F) of 14.4%[1].
Cell Assay	Western Blot Analysis[1] Cell Types: MDA-MB-231 cells Tested Concentrations: 0, 0.11, 0.33, 1, 3 μM. Incubation Duration: 3 days Experimental Results: Restored the protein levels of E-cadherin and N-cadherin to control levels . Cell Migration Assay [1] Cell Types: MDA-MB-231 cell Tested Concentrations: 0, 0.11, 0.33, 1, 3 μM. Incubation Duration: 24 h Experimental Results: Inhibited cell migration at 1 and 3 μM. Inhibited the invasion of MDA -MB-231 cells by 22.6, 34.8, 56.5, and 70.4% at the concentrations of 0.11, 0.33, 1.0, and 3.0 μM, respectively.
Animal Protocol	Animal/Disease Models: Xenograft model derived from highly metastatic 4T1 cells.[1] Doses: 50 or 100 mg/kg Route of Administration: Oral administration (po) Experimental Results: Suppressed 4T1 tumor growth with a tumor growth inhibition (TGI) of 78.0 and 95.9% at 50 and 100 mg/kg, respectively. Inhibited the phosphorylation of AXL. demonstrated that liver is one of the most common sites of breast cancer metastasis. Animal/Disease Models: Rats[1] Doses: 5 mg/kg (iv), 25 mg/ kg (po) Route of Administration: intravenous (iv) injection (iv), oral administration (po) Experimental Results: pharmacokinetic/PK parameters of AXL-IN-13 (Compound 6li). parameters T1/2 (h) Cmax (ng/mL) AUClast F (%) 5 mg/kg (iv) 3.31 12280.44 11684.24 25 mg/kg (po) 4.22 887.75 8410.21 14.4
References	[1]. Discovery of 3-Aminopyrazole Derivatives as New Potent and Orally Bioavailable AXL Inhibitors. J Med Chem. 2022 Nov 24;65(22):15374-15390.

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	1.5805 mL	7.9023 mL	15.8045 mL
	5 mM	0.3161 mL	1.5805 mL	3.1609 mL
	10 mM	0.1580 mL	0.7902 mL	1.5805 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.