TL02-59 diHCl is an orally bioactive, selective inhibitor of the Src-family kinase Fgr with IC50 of 0.03 nM. TL02-59 diHCl inhibits Src-family kinases Lyn and Hck with IC50 of 0.1 nM and 160 nM, respectively. TL02-59 diHCl effectively inhibits acute myeloid leukemia (AML) cell growth.

Physicochemical Properties

Molecular Formula	C32H35CLF3N5O4
Molecular Weight	646.10
Exact Mass	681.209
CAS #	2415263-06-6
Related CAS #	TL02-59;1315330-17-6
PubChem CID	146018965
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	11
Rotatable Bond Count	9
Heavy Atom Count	46
Complexity	923
Defined Atom Stereocenter Count	0
InChi Key	WLZPBFIIZXYWTD-UHFFFAOYSA-N
InChi Code	InChI=1S/C32H34F3N5O4.2ClH/c1-5-39-10-12-40(13-11-39)18-22-8-9-23(15-25(22)32(33,34)35)38-30(41)21-7-6-20(2)27(14-21)44-31-24-16-28(42-3)29(43-4)17-26(24)36-19-37-31;;/h6-9,14-17,19H,5,10-13,18H2,1-4H3,(H,38,41);2*1H
Chemical Name	3-(6,7-dimethoxyquinazolin-4-yl)oxy-N-[4-[(4-ethylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl]-4-methylbenzamide;dihydrochloride
Synonyms	TL0259 diHCl; TL02 59 diHCl
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	In TF-1 cells, TL02-59 diHClide (0.1-1000 nM; 6 hours) potently inhibits Fgr autophosphorylation, partially at 0.1-1 nM and fully above 10 nM. The range of 100–1000 nM inhibits Hck, Lyn, and Flt3 [1]. AML cell lines expressing this kinase with single-digit nM potency are inhibited in proliferation and undergo apoptosis by TL02-59 diHClide [1]. Primary AML bone marrow samples undergo growth stoppage due to TL02-59 dihydrochloride [1].
ln Vivo	TL02-59 (oral; 1 and 10 mg/kg; for three weeks) greatly reduces bone marrow involvement while totally eliminating AML cells in the spleen and peripheral blood of AML animal models [1]. When administered intravenously, TL02-59 has a t1/2 of 5.7 hours; when administered orally, it is 6.5 hours [1].
Cell Assay	Western Blot Analysis[1] Cell Types: TF-1 bone marrow cells Tested Concentrations: 0.1, 1, 10, 100, 1000 nM Incubation Duration: 6 hrs (hours) Experimental Results: Inhibition of Fgr autophosphorylation in TF-1 cells.
Animal Protocol	Animal/Disease Models: NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice with human MV4-11 AML cells [1] Doses: 1 and 10 mg/kg Route of Administration: Oral; for three weeks Experimental Results: Elimination of AML in mice AML cells in the spleen and peripheral blood in the model, while Dramatically inhibiting bone marrow involvement.
References	[1]. Selective Inhibition of the Myeloid Src-Family Kinase Fgr Potently Suppresses AML Cell Growth in Vitro and in Vivo. ACS Chem Biol. 2018 Jun 15;13(6):1551-1559.

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	1.5477 mL	7.7387 mL	15.4775 mL
	5 mM	0.3095 mL	1.5477 mL	3.0955 mL
	10 mM	0.1548 mL	0.7739 mL	1.5477 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.