Antitumor agent-61 (Compound 9b) is an Irinotecan (Ir) analogue and a potential antitumor agent. The IC50 of Antitumor agent-61 against tumor cells SK-OV-3, SK-OV-3/CDDP, U2OS, MCF-7, A549 and MG-63 were 0.92, 1.39, 1.75, 2.20, 3.05 and 3.23 μM respectively. Antitumor agent-61 causes apoptosis in SK-OV-3 cells through the mitochondrial pathway.

Physicochemical Properties

Molecular Formula	C54H63FN5O10P
Molecular Weight	992.08
CAS #	2408917-12-2
PubChem CID	162645608
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	14
Rotatable Bond Count	19
Heavy Atom Count	71
Complexity	2000
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

ln Vitro	Antineoplastic agent 61 (compound 6b) exhibited antiproliferative activity against A549, MCF-7, MG-63, U2OS, SK-OV-3, and SK-μM OV-3/CDDP cells, with IC50 values of 3.05, 2.20, 3.23, 1.75, 0.92, and 1.39[1]. At 150 μM, antitumor agent-61 (compound 6b) demonstrates some inhibitory effect against Topo I [1]. Through the mitochondrial route, antitumor agent 61 (compound 6b) (5-10 μM; 24 hours, SK-OV-3 cells) promotes apoptosis. dose-dependently raises ROS levels and lowers MMP levels [1].
ln Vivo	Average NPC tumor burden was dose-dependently reduced by Antitumor Agent-61 (Compound 6b) (10–20 mg/kg; ih; twice daily for 28 days; BALB/c nude mice with SK-OV-3 xenografts) [1].
Cell Assay	Cytotoxicity assay [1] Cell Types: SK-OV-3 Cell Tested Concentrations: 5.0 and 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: The percentage of apoptotic cells (including early and late apoptosis) increased from 21.11% (5 μM) to 32.27 % (10 μM), the cell apoptosis rate was Dramatically higher than that of Ir. Cell cycle analysis[1] Cell Types: SK-OV-3 Cell Tested Concentrations: 5.0 and 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: S phase percentage increased in a dose-dependent manner. Western Blot Analysis [1] Cell Types: SK-OV-3 cells Tested Concentrations: 5.0 and 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: The expression of anti-apoptotic protein Bcl-2 diminished, and the expression of pro-apoptotic proteins Bax and caspase increased.
Animal Protocol	Animal/Disease Models: SK-OV-3 xenograft BALB/c nude mice [1] Doses: 10 and 20 mg/kg Route of Administration: subcutaneous injection; 20 mg/kg. Twice a day for 28 days. Experimental Results: 10 mg/kg and 20 mg/kg inhibited tumor growth by 47.7% and 56.8% respectively, without affecting body weight or causing any obvious side effects.
References	[1]. Synthesis, mechanisms of action, and toxicity of novel aminophosphonates derivatives conjugated irinotecan in vitro and in vivo as potent antitumor agents. Eur J Med Chem. 2020 Mar 1;189:112067.

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	1.0080 mL	5.0399 mL	10.0798 mL
	5 mM	0.2016 mL	1.0080 mL	2.0160 mL
	10 mM	0.1008 mL	0.5040 mL	1.0080 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.