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

Physicochemical Properties

Molecular Formula	C22H31N5
Related CAS #	3,6-DMAD dihydrochloride;2226511-77-7
Appearance	Yellow to orange solid powder
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	3,6-DMAD hydrochloride is cytotoxic to MM cell lines (0–6 μM; 24 h; RPMI 8226 and MM1.R human MM cells)[1]. In HT1080 cells treated with Tg (0.3 μM) for 14 hours, 3,6-DMAD hydrochloride (0-30 μM) inhibits XBP1 splicing (XBP1s) in a dose-dependent manner[1]. The IRE1α endonuclease activity is inhibited by 3,6-DMAD hydrochloride (0.1-500 μM; 14 h; HT1080 cells treated with Tg(0.3 μM))[1]. IRE1α oligomerization and the development of IRE1α-GFP foci are inhibited by 3,6-DMAD hydrochloride (1-60 μM; 2 h; HEK293 cells)[1].
ln Vivo	In vivo suppression of XBP1 splicing has been seen in NOD Scid mice with RPMI 8226 xenograft when given 3,6-DMAD hydrochloride (10 mg/kg; ip; three times every 12 hours, for 84 hours)[1]. RPMI 8226 xenograft-producing multiple myeloma xenograft growth is inhibited in vivo by 3,6-DMAD hydrochloride (10 mg/kg; 24 h; ip; every 48 hours, for 12 days; NOD Scid mice)[1].
Cell Assay	Cell Viability Assay[1] Cell Types: RPMI 8226 and MM1.R human MM cells Tested Concentrations: 0, 0.5, 1, 2, 3, 4, 5 and 6 μM Incubation Duration: 24 hrs (hours) Experimental Results: Inhibited cell survival rate in a dose dependent manner. Western Blot Analysis[1] Cell Types: HT1080 cells treated with Tg(0.3 μM) Tested Concentrations: 0, 5, 10 and 30 μM Incubation Duration: 14 hrs (hours) Experimental Results: demonstrated XBP1s inhibition at as low as 0.5 μM.
Animal Protocol	Animal/Disease Models: NOD SCID (severe combined immunodeficient) mouse (4-6 weeks) with RPMI 8226 xenograft[1] Doses: 10 mg/kg Route of Administration: intraperitoneal (ip)injection; three times every 12 hrs (hours), for 84 hrs (hours) Experimental Results: Inhibited XBP1-luciferase activity in NOD SCID (severe combined immunodeficient) mouse with RPMI 8226 xenograft. Animal/Disease Models: NOD SCID (severe combined immunodeficient) mouse (4 -6 weeks) with RPMI 8226 xenograft[1] Doses: 10 mg/kg Route of Administration: intraperitoneal (ip)injection; every 48 hrs (hours), for 12 days Experimental Results: Inhibited tumor growth in NOD SCID (severe combined immunodeficient) mouse with RPMI 8226 xenograft.
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)

DMSO :~25 mg/mL

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.)