IRG1-IN-1 is an itaconic acid analogue. IRG1-IN-1 inhibits the activity of immune response gene 1 (IRG1). IRG1-IN-1 may be utilized in research on cancer, inflammation, and autoimmune diseases.

Physicochemical Properties

Molecular Formula	C18H15FO4
Molecular Weight	314.307708978653
Exact Mass	314.095
CAS #	2407652-42-8
PubChem CID	153387663
Appearance	White to off-white solid powder
LogP	3.4
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	6
Heavy Atom Count	23
Complexity	446
Defined Atom Stereocenter Count	0
SMILES	C(O)(=O)C(=CC1=CC=C(F)C=C1)C(CC1=CC=CC=C1)C(O)=O
InChi Key	DZXOXUPVMSOERA-WJDWOHSUSA-N
InChi Code	InChI=1S/C18H15FO4/c19-14-8-6-13(7-9-14)11-16(18(22)23)15(17(20)21)10-12-4-2-1-3-5-12/h1-9,11,15H,10H2,(H,20,21)(H,22,23)/b16-11-
Chemical Name	(3Z)-2-benzyl-3-[(4-fluorophenyl)methylidene]butanedioic acid
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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 6 (IRG1-IN-1; 0.5 mM; 2 mM) inhibits the release of TNFα and itaconic acid from human monocyte-derived macrophages (hMDMs) activated by lipopolysaccharide (LPS) [1]. C-IRG1-9 rat glioma cell growth is inhibited by IRG1-IN-1 (0.5 mM; 1 mM)[1]. TCR-activated hCD8+ T cell proliferation is enhanced by IRG1-IN-1 (10 nM)[1]. In CR-activated hCD8+ T cells, IRG1-IN-1(10μM) exhibits a reduction in the trimethylation of histone 3 at lysine 4 (H3K4me3)[1].
ln Vivo	In C57BL/6 mice, IRG1-IN-1 (compound 6) (ip; 0.2 mg/kg; 27 days) exhibits antitumor effect[1].
Cell Assay	Western Blot Analysis[1] Cell Types: TCR-activated hCD8+ T cells Tested Concentrations: 10 μM Incubation Duration: 24-72 h Experimental Results: diminished protein levels of histone 3 (H3). Cell Proliferation Assay[1] Cell Types: C-IRG1-9 rat glioma cells and TCR-activated hCD8+ T cells Tested Concentrations: 0.5 mM; 1 mM; 10 nM Incubation Duration: 48- 96 h Experimental Results: Inhibited the proliferation of C-IRG1-9 rat glioma cells and increased proliferation of TCR-activated hCD8+ T cells.
Animal Protocol	Animal/Disease Models: C57BL/6 mice[1] Doses: 0.2 mg/kg Route of Administration: IP; 27 days Experimental Results: Increased survival of C57BL/6 mice bearing mouse CT26 colorectal tumors. diminished intratumoral frequency of M-MDSCs in tumors.
References	[1]. Compositions and methods of using itaconic acid derivatives. Patent. US20210261495A1.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (318.16 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.95 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.95 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (7.95 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.1816 mL	15.9079 mL	31.8157 mL
	5 mM	0.6363 mL	3.1816 mL	6.3631 mL
	10 mM	0.3182 mL	1.5908 mL	3.1816 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.