Antitumor agent-115 (SS-12) is a potent in vitro antitumor compound with IC50 of 0.34 μM-24.14 μM against mouse breast cancer cell line 4T1 cells. Antitumor agent-115 can block the cell cycle of mouse breast cancer cell line 4T1, reduce mitochondrial membrane potential, and cause apoptosis. The IC50 for 4T1 cell viability is 8-25 μmol/L, and can be used for treating breast cancer diseases. Research.

Physicochemical Properties

Molecular Formula	C19H38CLNO2
Molecular Weight	347.96
CAS #	2759277-20-6
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	In 4T1 cells, antitumor agent-115 (SS-12) (1.25 μM, 10 μm, 40 μM, 24 hours) mostly causes apoptosis by stopping the cell cycle in the G0/G1 phase [1]. 4T1 cell migration and invasion are inhibited by antitumor agent-115 (15 μM, 30 μM, 60 μM, 24 hours) [1]. 4T1 cells undergo apoptosis when exposed to antitumor agent-115 (10 μM) for a full day [1].
ln Vivo	Antitumor agent-115 (SS-12) exhibits a good bioavailability in rats when administered orally (10 mg stachydrine/kg equivalency, 200 μL) [1]. In breast cancer model mice, antitumor agent-115 (0.1 mL/10 g, oral treatment) exhibits antitumor effects and enhances the morphological structure of the spleen [1].
Cell Assay	Cell Viability Assay[1] Cell Types: 4T1 cell Tested Concentrations: 1-128 μM Incubation Duration: 24, 48, 72 h Experimental Results: Inhibited 4T1 cells proliferation with IC50 value of 8-25 μM. Apoptosis Analysis[1] Cell Types: 4T1 cell Tested Concentrations: 1.25 μM, 10 μM, 40 μM Incubation Duration: 24 h Experimental Results: Increased apoptosis rates of 4T1 cell at concentration of 1.25 μM. Cell Cycle Analysis[1] Cell Types: 4T1 cell Tested Concentrations: 1.25 μM, 10 μM, 40 μM Incubation Duration: 24 h Experimental Results: Increased the proportion in the G0/G1 phase and diminished in the S and G2/M phases of 4T1 cells. Cell Migration Assay [1] Cell Types: 4T1 cell Tested Concentrations: 15 μM, 30 μM, 60 μM Incubation Duration: 24 h Experimental Results: Expanded the wound area and inhibited cell migration and invasion at concentration of 0.03 μM. Western Blot Analysis[1] Cell Types: 4T1 cell Tested Concentrations: 10 μM Incubation Duration: 24 h Experimental Results: Improved the cleaved caspase-3 expression and and Bax expression, but inhibited the anti-apoptosis protein Bcl2 expression in 4T1 cells.
Animal Protocol	Animal/Disease Models: Rats[1] Doses: 10 mg stachydrine/kg equivale, 200 μL Route of Administration: Oral administration (po) Experimental Results: demonstrated high systemic exposure with 6.10 μg/mL of Cmax value and 14.75 h μg/mL of AUC0-t value. Animal/Disease Models: Tumor-bearing mice[1] Doses: 0.2 mmol/kg Route of Administration: Oral administration (po) Experimental Results: Inhibitory rate was 51.47% ( P < 0.01) Displayed significant tumor cell shrinkage and larger void areas in tumor tissues demonstrated more weight loss than the model group and other treatment groups, increased of both AST and ALT levels and diminished spleen index of the tumor-bearing mice
References	[1]. Zeng H, et al. Synthesis, characterization and anti-breast cancer activities of stachydrine derivatives. Eur J Med Chem. 2023 Jul 26;259:115679.

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	2.8739 mL	14.3695 mL	28.7389 mL
	5 mM	0.5748 mL	2.8739 mL	5.7478 mL
	10 mM	0.2874 mL	1.4369 mL	2.8739 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.