Antitumor agent-70 (compound 8b) has anti-tumor effects and can cause apoptosis. Antitumor agent-70 inhibits multiple myeloma with IC50 of 0.12 μM. Antitumor agent-70 is a potential multi-target kinase inhibitor, especially for c-Kit.

Physicochemical Properties

Molecular Formula	C21H18N4O2
Molecular Weight	358.393224239349
Exact Mass	358.142
CAS #	2454133-88-9
PubChem CID	153531425
Appearance	Typically exists as solid at room temperature
LogP	3
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	5
Heavy Atom Count	27
Complexity	520
Defined Atom Stereocenter Count	0
SMILES	CC1=CC=C(C=C1)NC(=O)C2=CC=C(C=C2)/C=N/NC(=O)C3=CN=CC=C3
InChi Key	IXYMMBYRNJWPSX-YDZHTSKRSA-N
InChi Code	InChI=1S/C21H18N4O2/c1-15-4-10-19(11-5-15)24-20(26)17-8-6-16(7-9-17)13-23-25-21(27)18-3-2-12-22-14-18/h2-14H,1H3,(H,24,26)(H,25,27)/b23-13+
Chemical Name	N-[(E)-[4-[(4-methylphenyl)carbamoyl]phenyl]methylideneamino]pyridine-3-carboxamide
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	Compound 8b, or antitumor agent-70, demonstrates remarkable anti-proliferative properties, particularly against multiple myeloma cells (RPMI8226), at concentrations of 0–50 μM during a 24-hour period. Cell cycle arrest occurs in the G0/G1 phase when antineoplastic agent 70 (compound 8b) is applied (0-0.2 μM, 24 hours) [1]. Antitumor agent-70, also known as compound 8b, can cause cell apoptosis by encouraging the generation of reactive oxygen species (ROS) from cells, which in turn stops cell growth [1].
Cell Assay	Cell proliferation assay [1] Cell Types: Human myeloma cell line U266, human multiple myeloma cell line RPMI8226, human umbilical vein endothelial cells HUVEC Tested Concentrations: 0-50 μM Incubation Duration: 24 hrs (hours) Experimental Results: demonstrated for U266, RPMI8226 The IC50 values of anti-proliferative activity and HUVEC are 3.81 μM, 0.12 μM, and 12.09 μM respectively. Cell cycle analysis [1] Cell Types: RPMI8226 Tested Concentrations: 0-0.2 μM Incubation Duration: 24 hrs (hours) Experimental Results: The proportion of cells in G0/G1 phase increased Dramatically, and the proportion of cells in S phase and G2/M phase diminished Dramatically. Apoptosis analysis [1] Cell Types: RPMI8226 Tested Concentrations: 0-0.2 μM Incubation Duration: 24 hrs (hours) Experimental Results: The induced apoptosis rate increased with increasing concentration. The early apoptosis rate is higher than the late apoptosis rate.
References	[1]. Design, synthesis and biological evaluation of novel (E)-N-phenyl-4-(pyridine-acylhydrazone) benzamide derivatives as potential antitumor agents for the treatment of multiple myeloma (MM). Bioorg Chem. 2020 Oct;103:104189.

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	2.7903 mL	13.9513 mL	27.9026 mL
	5 mM	0.5581 mL	2.7903 mL	5.5805 mL
	10 mM	0.2790 mL	1.3951 mL	2.7903 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.