AMI-1 (AMI 1) disodium salt is a novel, cell-permeable and selective inhibitor of Histone Methyltransferase (HMT) with anti-inflammatory activity. It inhibits HMT with an IC50 of 3.0 μM and 8.8 μM for yeast Hmt1p and human PRMT1, respectively.

Physicochemical Properties

Molecular Formula	C21H14N2NA2O9S2
Molecular Weight	548.45
Exact Mass	547.993
CAS #	20324-87-2
Related CAS #	AMI-1 free acid;134-47-4
PubChem CID	88489
Appearance	Light brown to brown solid powder
LogP	5.164
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	2
Heavy Atom Count	36
Complexity	859
Defined Atom Stereocenter Count	0
InChi Key	MOUNHKKCIGVIDI-UHFFFAOYSA-L
InChi Code	InChI=1S/C21H16N2O9S2.2Na/c24-19-9-15(33(27,28)29)7-11-5-13(1-3-17(11)19)22-21(26)23-14-2-4-18-12(6-14)8-16(10-20(18)25)34(30,31)32;;/h1-10,24-25H,(H2,22,23,26)(H,27,28,29)(H,30,31,32);;/q;2*+1/p-2
Chemical Name	Disodium 7,7''-(carbonyldiimino)bis(4-hydroxynaphthalene-2-sulphonate)
Synonyms	AMI-1 disodium salt; AMI 1; AMI1; AMI-1
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, avoid exposure to moisture.
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	The in vitro methylation reactions carried out by the five recombinantly active PRMTs (PRMT1, -3, -4, and -6 as well as Hmt1p) can be inhibited by AMI-1[2]. AMI-1 inhibits type II PRMT5 in addition to type I PRMTs (PRMT1, 3, 4, and 6)[2]. AMI-1 does not compete with AdoMet for the binding site and selectively inhibits the methyltransferase activity of arginine in vitro, but not that of lysine[3]. Cellular proteins and GFP-Npl3 methylation are both inhibited by AMI-1[3]. In vitro, sarcoma in S180 and U2OS cells is inhibited by AMI-1 (0.6-2.4 mM; 48-96 hours) in a time- and dose-dependent manner[4]. AMI-1 (1.2–2.4 mM; 48–72 hours) induces apoptosis in cells, which lowers the viability of S180 cells[4].
ln Vivo	AMI-1 reduces S180 viability in vivo at a dose of 0.5 mg intraperitoneally every day for seven days[4]. ?In a tumor xenograft model, AMI-1 (0.5 mg; intratumorally; daily; for 7 days) downregulates PRMT5 but does not control PRMT7 expression[4]. ?In a tumor xenograft model, AMI-1 (0.5 mg; intratumorally; daily; for 7 days) reduces the levels of H4R3me2s and H3R8me2s[4].
Cell Assay	Cell Viability Assay[4] Cell Types: S180 cells, U2OS cells Tested Concentrations: 0.6 mM, 1.2 mM, 2.4 mM Incubation Duration: 48 hrs (hours), 72 hrs (hours), 96 hrs (hours) Experimental Results: Inhibited the cell viability. Increased the percentages of cells undergoing apoptosis.
Animal Protocol	Animal/Disease Models: 6-7 weeks old male Kunming mice (18-22 g), with S180 cells xenograft[4] Doses: 0.5 mg Route of Administration: Intratumorally, daily, for 7 days Experimental Results: diminished tumor weight.
References	[1]. Targeting protein arginine methyltransferase 5 inhibits colorectal cancer growth by decreasing arginine methylation of eIF4E and FGFR3. Oncotarget. 2015 Sep 8;6(26):22799-811. [2]. Arginine Methyltransferase inhibitor-1 Inhibits Sarcoma Viability in vitro and in vivo. Oncol Lett. 2018 Aug;16(2):2161-2166. [3]. Small Molecule Regulators of Protein Arginine Methyltransferases. J Biol Chem. 2004 Jun 4;279(23):23892-9.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (182.3 mM) Water:10 mg/mL (18.2 mM) Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.56 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 (4.56 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: PBS: 22mg/mL Solubility in Formulation 4: 100 mg/mL (182.33 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.8233 mL	9.1166 mL	18.2332 mL
	5 mM	0.3647 mL	1.8233 mL	3.6466 mL
	10 mM	0.1823 mL	0.9117 mL	1.8233 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.