| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
Inositol-requiring enzyme 1alpha (IRE1alpha) and X-Box Binding Protein 1 splicing (XBP1s). IRE1alpha is a sensor of endoplasmic reticulum (ER) stress. 3,6-DMAD hydrochloride is a potent IRE1alpha-XBP1s pathway inhibitor. It inhibits IRE1alpha oligomerization and its endoribonuclease (RNase) activity, blocking the splicing of XBP1 and thus inhibiting the downstream UPR, which can promote apoptosis in cancer cells.
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| 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].
In vitro, 3,6-DMAD hydrochloride inhibits XBP1 splicing in a dose-dependent manner in HT1080 cells treated with ER stress inducer thapsigargin (Tg). It also shows cytotoxicity against multiple myeloma (MM) cell lines RPMI 8226 and MM1.R, with an IC50 in the low micromolar range (0-6 microM; 24 h). This confirms its role in promoting cancer cell death. |
| 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].
In vivo, 3,6-DMAD hydrochloride suppresses multiple myeloma xenograft growth in NOD Scid mice. Administered intraperitoneally (i.p.) at 10 mg/kg every 48 hours for 12 days, it significantly inhibits tumor growth in the RPMI 8226 xenograft model. This suggests that targeting the IRE1alpha-XBP1 pathway is a valid therapeutic strategy for multiple myeloma. |
| Enzyme Assay |
A non-cell biochemical assay is performed to measure IRE1alpha RNase activity. Purified recombinant IRE1alpha protein is incubated with an RNA stem-loop substrate (XBP1 hairpin). The reaction is performed in the presence of varying concentrations of the test compound. The RNA fragments are separated by gel electrophoresis, and the inhibition of XBP1 splicing is quantified to determine the IC50.
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| 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. Cells (e.g., HT1080, RPMI 8226) are seeded in 96-well plates and treated with varying concentrations of 3,6-DMAD hydrochloride for 24-72 hours. Cell viability is measured using an MTT or CellTiter-Glo assay. XBP1 splicing is confirmed by RT-PCR, where the shift from unspliced (u) to spliced (s) XBP1 is blocked by the inhibitor. |
| 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. In vivo efficacy is evaluated in a RPMI 8226 multiple myeloma xenograft model in NOD Scid mice. Mice are injected subcutaneously with tumor cells. When tumors reach a certain size, the compound is administered intraperitoneally (IP) at a dose of 10 mg/kg every 48 hours for 12 days. Tumor volume is measured by calipers twice weekly. Body weight is monitored to assess toxicity. |
| ADME/Pharmacokinetics |
3,6-DMAD hydrochloride has a molecular weight of 400.87. It is soluble in DMSO. For in vivo administration (IP), it can be formulated in a vehicle such as 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline. Detailed PK parameters such as half-life, Cmax, and bioavailability are not publicly available for this research compound.
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| Toxicity/Toxicokinetics |
In animal studies, 3,6-DMAD hydrochloride is well-tolerated at efficacious doses (10 mg/kg, IP). No significant body weight loss or overt toxicity was reported. The compound is for research use only. Comprehensive toxicity studies, including genotoxicity and chronic toxicity, have not been published.
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| References |
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| Additional Infomation |
3,6-DMAD hydrochloride is a research-grade chemical tool. It is a potent IRE1alpha-XBP1s pathway inhibitor used to study the unfolded protein response in cancer. The IRE1alpha-XBP1 pathway is a validated target for treating plasma cell malignancies. This product is for research use only and is not FDA-approved for human therapy. Key references describe its role in multiple myeloma.
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| Molecular Formula |
C22H31N5
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| Related CAS # |
3,6-DMAD dihydrochloride;2226511-77-7
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| Appearance |
Yellow to orange solid powder
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO :~25 mg/mL
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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