Antitumor agent-19 is an antitumor agent that targets tumor-associated macrophages (TAMs) and could be utilized as a potent modulator of TAMs. The EC50s of Antitumor agent-19 in RAW 264.7 cells and BMDM cells were 17.18 µM and 18.87 µM, respectively.

Physicochemical Properties

Molecular Formula	C24H21CLF3N5O
Molecular Weight	487.90465426445
Exact Mass	487.138
CAS #	2379727-90-7
PubChem CID	139600306
Appearance	Typically exists as solid at room temperature
LogP	5
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	3
Heavy Atom Count	34
Complexity	742
Defined Atom Stereocenter Count	0
InChi Key	RTILPKXOEYEQSE-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H21ClF3N5O/c1-33-12-13(11-29-33)16-3-2-4-21-22(16)17-9-14(6-8-20(17)32-21)30-23(34)31-15-5-7-19(25)18(10-15)24(26,27)28/h2-5,7,10-12,14,32H,6,8-9H2,1H3,(H2,30,31,34)
Chemical Name	1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[5-(1-methylpyrazol-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]urea
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

Targets	tumor-associated macrophages (TAMs)[1]
ln Vitro	Antitumor agent-19 (Compound 23a; 10-50 µM; 1 hour; RAW 264.7 cells) successfully promotes the expression of M1-phenotype macrophage markers (IL-1β, TNF-α, iNOS, and IL-12α) in RAW 264.7 cells in a dose-dependent manner over the course of an hour. It has been observed that antitumor agent-19 repolarizes TAMs from M1 (anti-tumor) and M2 (pro-tumor) in a dose-dependent manner[1].
ln Vivo	The LLC mouse model's tumor growth can be significantly inhibited by antitumor agent-19 (Compound 23a; 5 mg/kg; intraperitoneal injection; every other day; for 30 days; C57BL/6 female mice). Furthermore, the combined use of anti-PD-1 antibody and antitumor agent-19 provides more potent antitumor effects than using either one alone in vivo[1].
Cell Assay	RT-PCR[1] Cell Types: RAW 264.7 cells Tested Concentrations: 10 µM, 20 µM, or 50 µM Incubation Duration: 1 hour Experimental Results: Induced the expression of M1-phenotype macrophage markers (IL-1β, TNF-α, iNOS and IL-12α) in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: Six-to eightweeks old C57BL/6 female mice with lewis lung cancer cells (LLC)[1] Doses: 5 mg/kg Route of Administration: intraperitoneal (ip) injection; every other day; for 30 days Experimental Results: Remarkably inhibited tumor growth of the LLC mouse model.
References	[1]. Pei H, et al. iscovery of potent ureido tetrahydrocarbazole derivatives for cancer treatments through targeting tumor-associated macrophages. Eur J Med Chem. 2019 Sep 28;183:111741.

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	2.0496 mL	10.2480 mL	20.4960 mL
	5 mM	0.4099 mL	2.0496 mL	4.0992 mL
	10 mM	0.2050 mL	1.0248 mL	2.0496 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.