KIN59 (5'-O-Tritylinosine) is a potent allosteric inhibitor of thymidine phosphorylase. KIN59 inhibits FGF2-stimulated cell growth. KIN59 inhibits the expression of p-FGFR1 and P-Akt in cells stimulated by FGF2 (10 ng/mL). KIN59 has anticancer effect.

Physicochemical Properties

Molecular Formula	C29H26N4O5
Molecular Weight	510.54
Exact Mass	510.19
CAS #	4152-77-6
PubChem CID	135526599
Appearance	White to off-white solid powder
LogP	2.747
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	7
Heavy Atom Count	38
Complexity	793
Defined Atom Stereocenter Count	4
SMILES	C1=CC=C(C=C1)C(C2=CC=CC=C2)(C3=CC=CC=C3)OC[C@@H]4[C@H]([C@H]([C@@H](O4)N5C=NC6=C5N=CNC6=O)O)O
InChi Key	YEPZMZDCXFRHCL-ZYWWQZICSA-N
InChi Code	InChI=1S/C29H26N4O5/c34-24-22(38-28(25(24)35)33-18-32-23-26(33)30-17-31-27(23)36)16-37-29(19-10-4-1-5-11-19,20-12-6-2-7-13-20)21-14-8-3-9-15-21/h1-15,17-18,22,24-25,28,34-35H,16H2,(H,30,31,36)/t22-,24-,25-,28-/m1/s1
Chemical Name	9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(trityloxymethyl)oxolan-2-yl]-1H-purin-6-one
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	For GM7373 cells treated with FGF2 (30 ng/mL) and PBS (10%), respectively, KIN59 (0-100 µM; 24 h) suppresses cell growth with IC50 values of 5.8 and 63 µM [1]. In FGFR1-overexpressing GM7373-FGFR1 cells stimulated by FGF2 (10 ng/mL), KIN59 (60 µM; 30 min) suppresses the production of P-Akt and p-FGFR1 [1]. KIN59 has IC50 values of 44 μM and 67 μM, respectively, which indicate its ability to inhibit human thymidine phosphorylase (TPase) and recombinant bacteria (E. coli) [2].
ln Vivo	In mice, anticancer activity is demonstrated by KIN59 (15 mg/kg; subcutaneous injection; twice daily beginning on day 2 for 20 days) [1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: GM7373 cells Tested Concentrations: 0-100 µM Incubation Duration: 24 h Experimental Results: Inhibited FGF2 (30 ng /mL)-induced proliferation of bovine macrovascular endothelial GM7373 cells in a dose-dependent manner with IC50 values of 5.8, 63 µM for FGF2 and PBS, respectively. Western Blot Analysis[1] Cell Types: GM7373-FGFR1, GM7373-VEGFR2 cells Tested Concentrations: 60 µM Incubation Duration: 30 min Experimental Results: Inhibited FGFR1 phosphorylation and Akt activation triggered by FGF2 in FGFR1-overexpressing GM7373-FGFR1 cells, demonstrated minor inhibitors on VEGF-mediated VEGFR2 phosphorylation and Akt activation in GM7373 cells overexpressing VEGFR2.
Animal Protocol	Animal/Disease Models: Eightweeks old female, 25 g, athymic, nude nu/nu (nude) mice (F2T-luc2.9 cells)[1] Doses: 15 mg/kg Route of Administration: Sc; twice (two times) daily from day 2 (one time/day during the weekend) at a site distant from the tumor (inoculation) site for 20 days Experimental Results: Caused a significant inhibition in the rate of tumor growth.
References	[1]. The thymidine phosphorylase inhibitor 5'-O-tritylinosine (KIN59) is an antiangiogenic multitarget fibroblast growth factor-2 antagonist. Mol Cancer Ther. 2012 Apr;11(4):817-29. [2]. Thymidine phosphorylase is noncompetitively inhibited by 5'-O-trityl-inosine (KIN59) and related compounds. Nucleosides Nucleotides Nucleic Acids. 2006;25(9-11):975-80.

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	1.9587 mL	9.7936 mL	19.5871 mL
	5 mM	0.3917 mL	1.9587 mL	3.9174 mL
	10 mM	0.1959 mL	0.9794 mL	1.9587 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.