CYP4Z1-IN-1 (compound 7c) is a potent inhibitor of CYP4Z1 with IC50 of 41.8 nM. CYP4Z1-IN-1 reduces breast cancer stem cell marker expression, spheroid formation and metastasis capacity, and tumor initiation in a concentration-dependent fashion in vitro & in vivo.

Physicochemical Properties

Molecular Formula	C13H18N2O3
Molecular Weight	250.3
Exact Mass	250.13
Elemental Analysis	C, 62.38; H, 7.25; N, 11.19; O, 19.18
CAS #	2760611-38-7
PubChem CID	166642480
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	8
Heavy Atom Count	18
Complexity	261
Defined Atom Stereocenter Count	0
InChi Key	HJEKWTVRPSQJRV-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H18N2O3/c1-2-18-13(16)5-3-4-11-6-8-12(9-7-11)14-10-15-17/h6-10,17H,2-5H2,1H3,(H,14,15)
Chemical Name	ethyl (E)-4-(4-(N'-hydroxyformimidamido)phenyl)butanoate
Synonyms	compound 7c; CYP4Z1-IN-1; CYP4Z1-IN 1; CYP4Z1 IN-1; CYP4Z1 Inhibitor 7c; CYP4Z1-IN-7c; GLXC-26590;
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	CYP4Z1-IN-1 (compound 7c) exhibits anti-proliferative action against breast cancer stem cells (CSCs) at an IC50 of 483 ± 2.5 nM[1]. In breast cancer cells, CYP4Z1-IN-1 (0.8-51.2 μM, 24 h) refluxes the expression of stem cell markers (P-gp, Oct3/4, Nanog, ALDH1A1, and Sox2) [1]. Breast cancer cells can migrate sparsely when exposed to CYP4Z1-IN-1 (0.8–51.2 μM, 24-48 hours).
ln Vivo	CYP4Z1-IN-1 (compound 7c) (2000 mg/kg, recently, for 7 days) has no obvious toxicity and weight loss in mice [1]. CYP4Z1-IN-1 (12.8 μM, 72 h treatment of MCF-7 and MDA-MB-231 cells, then subcutaneous tumor cells inguinal breast) can induce tumescent cell initiation [1].
Cell Assay	Western Blot Analysis[1] Cell Types: MCF-7 and MDA-MB-231 Cell Tested Concentrations: 0.8 μM, 3.2 μM, 12.8 μM, 51.2 μM Incubation Duration: 24 h Experimental Results: Significant inhibition of protein and solubility [1]. The expression of stemness markers (P-gp, Oct3/4, Nanog, ALDH1A1, and Sox2) in MCF-7 cells was concentration-dependent.
Animal Protocol	Animal/Disease Models: Mice [1] Doses: 2000 mg/kg Route of Administration: Orally for 7 days Experimental Results: Compound 7c is shown to be quite safe; no obvious toxicity and weight loss were observed. Animal/Disease Models: BALB/c-nude mice (3-4 weeks old, female) [1] Doses: 12.8 μM Route of Administration: MCF-7 and MDA-MB-231 cells were pretreated with 7c (12.8 μM) for 72 hrs (hrs (hours)) and then Implanted subcutaneously (sc) (sc) into the mouse inguinal mammary gland Experimental Results: Blocks the tumor-initiating ability of breast cancer cells.
References	[1]. Identification of a Novel Potent CYP4Z1 Inhibitor Attenuating the Stemness of Breast Cancer Cells through Lead Optimization. J Med Chem. 2022 Dec 8;65(23):15749-15769.

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	3.9952 mL	19.9760 mL	39.9521 mL
	5 mM	0.7990 mL	3.9952 mL	7.9904 mL
	10 mM	0.3995 mL	1.9976 mL	3.9952 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.