3,6-DMAD diHCl is an acridine analogue and a potent inhibitor of the IRE1α-XBP1s pathway. 3,6-DMAD diHCl promotes IL-6 secretion through the IRE1α-XBP1s pathway. 3,6-DMAD diHCl inhibits IRE1α oligomerization and endoribonuclease (RNase) activity. 3,6-DMAD diHCl may be used in cancer-related research.

Physicochemical Properties

Molecular Formula	C22H32CLN5
Molecular Weight	401.98
Exact Mass	437.211
CAS #	2226511-77-7
Related CAS #	3,6-DMAD hydrochloride
PubChem CID	165412452
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	7
Heavy Atom Count	29
Complexity	420
Defined Atom Stereocenter Count	0
InChi Key	QFTZLLNIVBXXJG-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H31N5.2ClH/c1-25(2)13-7-12-23-22-18-10-8-16(26(3)4)14-20(18)24-21-15-17(27(5)6)9-11-19(21)22;;/h8-11,14-15H,7,12-13H2,1-6H3,(H,23,24);2*1H
Chemical Name	9-N-[3-(dimethylamino)propyl]-3-N,3-N,6-N,6-N-tetramethylacridine-3,6,9-triamine;dihydrochloride
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	3,6-DMAD diHClide (0-6 μM; 24 hours; RPMI 8226 and MM1.R human MM cells) is MM cell line-cytotoxic [1]. In HT1080 cells treated with Tg (0.3 μM), 3,6-DMAD dihydrochloride (0-30 μM; 14 hours) inhibits XBP1 splicing (XBP1s) in a dose-dependent manner [1]. IRE1α endonuclease activity is inhibited by 3,6-DMAD dihydrochloride (0.1-500 μM; 14 hours; HT1080 cells treated with Tg (0.3 μM)) [1]. Inhibiting IRE1α oligomerization and IRE1α-GFP foci formation, 3,6-DMAD dihydrochloride (1-60 μM; 2 hours; HEK293 cells) is one way to treat this [1].
ln Vivo	In vivo XBP1 splicing is inhibited by 3,6-DMAD dihydrochloride (10 mg/kg; i.p.; administered three times every 12 hours for 84 hours) in NOD Scid mice bearing RPMI 8226 xenografts [1]. Multiple myeloma xenografts in vivo Graft growth is inhibited by 3,6-DMAD dihydrochloride (10 mg/kg; 24 hours; i.p.; every 48 hours for 12 days; NOD Scid mice using RPMI 8226 xenografts)[1].
Cell Assay	Cell Viability Assay[1] Cell Types: RPMI 8226 and MM1.R Human MM Cell Tested Concentrations: 0, 0.5, 1, 2, 3, 4, 5 and 6 μM Incubation Duration: 24 hrs (hours) Experimental Results: Single dose inhibition of cell viability dependent Way. Cell viability assay[1] Cell Types: 0, 5, 10 and 30 μM Tested Concentrations: 0, 0.5, 1, 2, 3, 4, 5 and 6 μM 24 hrs (hours) Incubation Duration: 14 hrs (hours) Experimental Results: Shows XBP1s inhibition as low as 0.5μM .
Animal Protocol	Animal/Disease Models: NOD SCID (severe combined immunodeficient) mouse (4-6 weeks), RPMI 8226 Grafts inhibit XBP1-luciferase activity in NOD SCID (severe combined immunodeficient) mouse. Animal/Disease Models: NOD SCID (severe combined immunodeficient) mouse (4-6 weeks), RPMI 8226 xenografts [1] Doses: 10 mg/kg Route of Administration: intraperitoneal (ip) injection; every 48 hrs (hrs (hours)) for 12 days Experimental Results: Using RPMI 8226 xenografts The agent inhibits tumor growth in NOD SCID (severe combined immunodeficient) mouse.
References	[1]. Acridine Derivatives as Inhibitors of the IRE1α-XBP1 Pathway Are Cytotoxic to Human Multiple Myeloma. Mol Cancer Ther. 2016 Sep;15(9):2055-65. [2]. SHP-2 specific deletion in macrophages accelerates pathological cardiac hypertrophy through promoting IRE1α-XBP1s pathway regulated by IL-6 secretion. Research Article. 2022 May 3.

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	2.4877 mL	12.4384 mL	24.8769 mL
	5 mM	0.4975 mL	2.4877 mL	4.9754 mL
	10 mM	0.2488 mL	1.2438 mL	2.4877 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.