Anticancer agent 43 is a novel anticancer agent that induces apoptosis by caspase 3, PARP1, and Bax dependent mechanisms. Also induces DNA damage.

Physicochemical Properties

Molecular Formula	C14H9FN2O3S2
Molecular Weight	336.361263990402
Exact Mass	336.003
CAS #	2470015-35-9
PubChem CID	163322351
Appearance	Light yellow to yellow solid powder
LogP	2.3
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	3
Heavy Atom Count	22
Complexity	630
Defined Atom Stereocenter Count	0
SMILES	COC(=O)C\1=NC2=C(/C1=C\C3=C(NC(=S)S3)O)C=C(C=C2)F
InChi Key	GBWAVAUPOYYLNO-VMPITWQZSA-N
InChi Code	InChI=1S/C14H9FN2O3S2/c1-20-13(19)11-8(5-10-12(18)17-14(21)22-10)7-4-6(15)2-3-9(7)16-11/h2-5,18H,1H3,(H,17,21)/b8-5+
Chemical Name	methyl (3E)-5-fluoro-3-[(4-hydroxy-2-sulfanylidene-3H-1,3-thiazol-5-yl)methylidene]indole-2-carboxylate
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 43 (compound 3a) exhibits selectivity for human tumor cells (SI50=28.94)[1]. HepG2 cells are exposed to anticancer agent 43 (45 µM) for 24 hours, which causes apoptosis via caspase 3, PARP1, and Bax-dependent pathways [1]. HepG2 cells undergo the G1/S phase transition without being affected by anticancer drug 43 (45 µM, 24 hours) [1]. HCT116 cells (tail DNA = 16.1%, OTM = 3.7), MCF-7 cells, HepG2 cells (tail DNA = 26.2%, OTM = 13.2), and Balb/DNA damagec 3T3 cells (tail DNA = 8.4%, OTM = 3.5) were all exposed to anticancer agent 43 (0.7, 45, and 55 µM) [1].
Cell Assay	Cytotoxicity assay[1] Cell Types: HepG2, MCF-7, HCT116, HeLa, A549, WM793, THP-1, HaCaT, Balb/c3T3 Cell Tested Concentrations: 0, 1, 10, 100 µM Incubation Duration: 72 hrs (hours) Experimental Results: It has cytotoxic effects on HepG2, MCF-7, HCT116, HeLa, and A549 cells, with a GI50 of 12.1, 0.7, 0.8, 49.3, and 9.7 µM. It has low toxicity to WM793, THP-1, HaCaT, and Balb/c 3T3 cells, with a GI50 of 80.4, 62.4, 98.3, 40.8 µM respectively. Apoptosis analysis[1] Cell Types: HepG2 Cell Tested Concentrations: 45 µM Incubation Duration: 24 hrs (hours) Experimental Results: Apoptosis in HepG2 cells was induced through caspase 3, PARP1 and Bax-dependent pathways. Western Blot Analysis [1] Cell Types: HCT116, MCF-7 Cell Tested Concentrations: 0.7 µM Incubation Duration: 24 h Experimental Results: Cdk2 protein expression was diminished in HCT116 and MCF-7 cells. Cell cycle analysis[1] Cell Types: HepG2 cells Tested Concentrations: 45 µM Incubation Duration: 24 hrs (hours) Experimental Results: No effect on the G1/S phase transition of HepG2 cells.
References	[1]. Synthesis of novel indole-thiazolidinone hybrid structures as promising scaffold with anticancer potential. Bioorg Med Chem. 2021; 50:116453.

Solubility Data

Solubility (In Vitro)

DMSO : ~125 mg/mL (~371.63 mM)

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	2.9730 mL	14.8650 mL	29.7301 mL
	5 mM	0.5946 mL	2.9730 mL	5.9460 mL
	10 mM	0.2973 mL	1.4865 mL	2.9730 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.