Canfosfamide (TLK-286) is a glutathione analog prodrug that is activated by glutathione S-transferase P1-1 and causes apoptosis. Canfosfamide also inhibits the catalytic kinase activity of DNA-dependent protein kinase (DNA-PK). Canfosfamide produces an anti-cancer alkylating agent and glutathione analogue upon activation. Canfosfamide may be utilized in the study of malignant tumors.

Physicochemical Properties

Molecular Formula	C26H40N5O10PSCL4
Molecular Weight	787.474100000001
Exact Mass	785.099
CAS #	158382-37-7
Related CAS #	439943-59-6 (HCl);158382-37-7;
PubChem CID	133045
Appearance	Typically exists as solid at room temperature
Density	1.484g/cm3
Boiling Point	939.8ºC at 760mmHg
Flash Point	522.2ºC
Vapour Pressure	0mmHg at 25°C
Index of Refraction	1.587
LogP	4.267
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	14
Rotatable Bond Count	24
Heavy Atom Count	47
Complexity	1150
Defined Atom Stereocenter Count	3
SMILES	O=C(O)[C@H](NC([C@@H](NC(CC[C@H](N)C(O)=O)=O)CS(=O)(CCOP(N(CCCl)CCCl)(N(CCCl)CCCl)=O)=O)=O)C1=CC=CC=C1
InChi Key	VESKBLGTHHPZJF-QNWVGRARSA-N
InChi Code	InChI=1S/C26H40Cl4N5O10PS/c27-8-12-33(13-9-28)46(42,34(14-10-29)15-11-30)45-16-17-47(43,44)18-21(32)24(37)35(22(36)7-6-20(31)25(38)39)23(26(40)41)19-4-2-1-3-5-19/h1-5,20-21,23H,6-18,31-32H2,(H,38,39)(H,40,41)/t20-,21-,23+/m0/s1
Chemical Name	(2S)-2-amino-5-[[(2R)-2-amino-3-[2-[bis[bis(2-chloroethyl)amino]phosphoryloxy]ethylsulfonyl]propanoyl]-[(R)-carboxy(phenyl)methyl]amino]-5-oxopentanoic acid
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	IC50: ~1 μM (DNA-PK)[3]
ln Vitro	Canfosfamide (TLK-28) has an IC50 value of less than 1 µM and inhibits the catalytic kinase activity of purified DNA-dependent protein kinase (DNA-PK), but it does not directly harm DNA very much[2]. In a dose-dependent manner, canfosfamide (TER286) inhibits MCF-7, NIH-3T3, and M7609[3].
ln Vivo	Canfosfamide (TER286) shows a more potent antitumor effect with increased dosing[3].
Cell Assay	Cell Proliferation Assay[3] Cell Types: MCF-7, NIH-3T3 and M7609 Tested Concentrations: 0-100 μM Incubation Duration: Co-incubated for 2 h then removed and incubated for 5 days in MCF-7; co-incubated for 10 days in NIH-3T3; co-incubated for 48 h in M7609 Experimental Results: Inhibited these cancer cell lines in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: BALB/c nude mice (sc with tumors of M7609, MX -1 human breast tumor, lung tumor[3] Doses: 150 mg/kg for M7609 xenografts; 400 mg/kg or 200 mg/kg for other xenografts Route of Administration: iv, single dosage for M7609 xenograft; ip, single dosage for other xenografts ; ip, daily for 5 days Experimental Results: The best response to TER286 was for the MX-1 human breast tumor treated with an aggressive regimen (daily for 5 days), under which nearly all of the tumors were either severely growth inhibited or substantially regressed .
References	[1]. Mechanism of glutathione transferase P1-1-catalyzed activation of the prodrug canfosfamide (TLK286, TELCYTA). Biochemistry. 2013 Nov 12;52(45):8069-78. [2]. Tew KD. TLK-286: a novel glutathione S-transferase-activated prodrug. Expert Opin Investig Drugs. 2005 Aug;14(8):1047-54. [3]. Tumor efficacy and bone marrow-sparing properties of TER286, a cytotoxin activated by glutathione S-transferase. Cancer Res. 1998 Jun 15;58(12):2568-75.
Additional Infomation	See also: Canfosfamide (annotation moved to).

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	1.2699 mL	6.3494 mL	12.6989 mL
	5 mM	0.2540 mL	1.2699 mL	2.5398 mL
	10 mM	0.1270 mL	0.6349 mL	1.2699 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.