Anticancer agent 205 (compound 9) is an effective anticancer agent. Anticancer agent 205 binds to the G4-mtDNA target and inhibits mtDNA (mitochondrial genome) replication, transcription, and translation. Anticancer agent 205 causes mitochondrial dysfunction, increases ROS production, induces DNA damage and cellular senescence. Anticancer agent 205 induces apoptosis and cell cycle arrest in the G0/G1 phase. Anticancer agent 205 has the potential to study colorectal cancer.

Physicochemical Properties

Molecular Formula	C52H60I2N4
Molecular Weight	994.87
Appearance	Typically exists as solid at room temperature
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	Anticancer agent 205 (compound 9) (4 µM; 1 h) interacts with G4-mtDNA in HCT116 cells[1]. Anticancer agent 205 (0, 1, 2, 4 µM; 48 h) reduces the mRNA levels of ND2 and ND5 in HCT116 and HFF1 cells[1]. Anticancer agent 205 (0, 1, 2, 4 µM; 48 h) reduces the protein levels of ND3, ND4, ND6, COX1, COX2, COX3, CYTB, ATP6, and ATP8 in HCT116 cells[1]. Anticancer agent 205 (0-4 µM) increases ROS levels in a concentration-dependent manner[1]. Anticancer agent 205 (0-4 µM; 72 h) induces DNA damage and cell senescence[1].
ln Vivo	Anticancer agent 205 (5 mg/kg; iv; every 2 days for 16 days) inhibits tumor growth in the HCT116 xenograft mouse model [1].
Cell Assay	Cell Cytotoxicity Assay[1] Cell Types: HCT116, LoVo, HeLa, HepG2, PANC-1, MDA-MB-231, HFF1, BJ Tested Concentrations: 0-40 µM Incubation Duration: 48 h Experimental Results: Showed cytotoxicity for HCT116, LoVo, HeLa, HepG2, PANC-1, MDA-MB-231, HFF1, BJ cells with IC50s of 3.4, 13.4, 23.5, >40, 24.8, 34.6, 32.7, >40 µM, respectively. Western Blot Analysis[1] Cell Types: HCT116 Tested Concentrations: 0, 1, 2, 4 µM Incubation Duration: 48 h Experimental Results: Showed that the protein levels of ND3, ND4, ND6, COX1, COX2, COX3, CYTB, ATP6, and ATP8 were decreased markedly. RT-PCR[1] Cell Types: HCT116, HFF1 Tested Concentrations: 0, 1, 2, 4 µM Incubation Duration: 48 h Experimental Results: Caused a significant reduction of mRNA levels for ND1, ND2, ND3, ND4, ND4L, ND5, COX2, ATP6, and ATP8 in HCT116, only ND2 and ND5 showed a significant reduction in HFF1 cells. Cell Cycle Analysis[1] Cell Types: HCT116 Tested Concentrations: 0, 1, 2, 4 µM Incubation Duration: 48 h Experimental Results: Induced cell cycle arrested in the G0/G1 phase with The percentage of the G0/G1 phase is increased from 50.66 to 66.01%, in a concentration-dependent manner, induces apoptosis by increases the expression of cleaved-caspase3 and the apoptosis cell increased from 2.64% to 27.4%.
Animal Protocol	Animal/Disease Models:male Balb/c nude mice (HCT116 cells)[1]. Doses: 5 mg/kg Route of Administration: I.v.; every 2 days for 16 days Experimental Results: Significantly inhibited the growth of tumors with the tumor growth reduced by 70% approximately.
References	[1]. Mitochondria-Selective Dicationic Small-Molecule Ligand Targeting G-Quadruplex Structures for Human Colorectal Cancer Therapy. J Med Chem. 2024 Apr 25;67(8):6292-6312.

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	1.0052 mL	5.0258 mL	10.0516 mL
	5 mM	0.2010 mL	1.0052 mL	2.0103 mL
	10 mM	0.1005 mL	0.5026 mL	1.0052 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.