| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Negative control for MZ1 (BRD4-targeted PROTAC)
Not applicable. cis-MZ 1 hydrate is an inactive negative control for a PROTAC molecule; it has no significant binding affinity for the von Hippel-Lindau (VHL) E3 ubiquitin ligase complex. The active PROTAC MZ 1 binds to both BRD4 (via its bromodomain ligand) and VHL (via its ligand), thereby inducing ubiquitination and subsequent proteasomal degradation of BRD4. The cis-isomer does not engage VHL due to its altered stereochemistry and thus cannot recruit the E3 ligase to BRD4, making it an inactive control. |
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| ln Vitro |
cis-MZ 1 hydrate is a negative control for the BRD4-targeted PROTAC MZ 1. MZ 1 is a combination of the von Hippel-Lindau (VHL) ligand and a BRD4 ligand. cis-MZ 1 does not have significant VHL binding affinity and therefore does not induce BRD4 degradation. It is used as an inactive control to differentiate target-specific PROTAC activity from nonspecific effects of the ligand fragments. The active compound MZ 1 binds to BRD4 BD1/2 with Kd values of 382 nM and 120 nM, respectively, and induces BRD4 degradation via the proteasome.
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| ln Vivo |
No in vivo data is relevant for cis-MZ 1 hydrate, as it is an inactive control compound. In vivo efficacy studies are performed with the active PROTAC MZ 1. In mouse xenograft models, MZ 1 has been shown to reduce BRD4 protein levels and inhibit tumor growth. cis-MZ 1 is used in such studies as a negative control to confirm that the antitumor effects observed with MZ 1 are specifically due to BRD4 degradation and not to off-target effects of the component ligands or the linker.
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| Enzyme Assay |
Not applicable. cis-MZ 1 hydrate is an inactive control compound and is not used in non-cellular biochemical assays for target binding. A typical BRD4 binding assay for the active compound MZ 1: Recombinant BRD4 BD1 or BD2 is incubated with a fluorescently labeled BET inhibitor probe and varying concentrations of MZ 1 or cis-MZ 1 in binding buffer (50 mM HEPES pH 7.5, 150 mM NaCl, 0.1% BSA) at room temperature for 30-60 minutes. Fluorescence polarization (FP) or TR-FRET is measured. Kd values are calculated (MZ 1: Kd = 382 nM for BD1, 120 nM for BD2). cis-MZ 1 shows no significant binding.
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| Cell Assay |
Standard cellular degradation assay for PROTAC activity: HEK293T or HeLa cells are seeded in 6-well plates. The next day, cells are treated with varying concentrations of MZ 1 (active) or cis-MZ 1 (inactive control) (0.1-1000 nM) for 4-16 hours. Cells are lysed in RIPA buffer containing protease inhibitors. BRD4 protein levels are measured by Western blot using an anti-BRD4 antibody. beta-actin or GAPDH is used as a loading control. The DC50 (half-maximal degradation concentration) is calculated for MZ 1. cis-MZ 1 should show no reduction in BRD4 levels compared to vehicle control, confirming its use as a negative control.
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| Animal Protocol |
No specific animal protocol is provided. In mouse xenograft studies using the active PROTAC MZ 1, cis-MZ 1 is used as a negative control. Female BALB/c nude mice (6-8 weeks old) are injected subcutaneously with 5×10⁶ cancer cells (e.g., 22Rv1 prostate cancer cells). When tumors reach ~150 mm3, mice are randomized into groups (n=8/group) and treated with MZ 1 (e.g., 50 mg/kg, IP), cis-MZ 1 (50 mg/kg, IP), or vehicle control, once daily for 14-21 days. Tumor volume is measured twice weekly. Tumors are harvested for BRD4 IHC and Western blot. Only MZ 1-treated mice should show tumor growth inhibition and BRD4 degradation.
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| ADME/Pharmacokinetics |
No PK data is provided for cis-MZ 1 hydrate. As an inactive control compound, its pharmacokinetic properties are not studied. The active PROTAC MZ 1 has a molecular weight of 681.8 and is typically administered intraperitoneally. MZ 1 has a half-life of approximately 1-2 hours in mice. The compound is rapidly absorbed after IP administration and is metabolized by liver CYP450 enzymes. cis-MZ 1 is expected to have a similar PK profile to MZ 1, but this is not routinely characterized.
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| Toxicity/Toxicokinetics |
cis-MZ 1 hydrate is an inactive control compound and is not associated with pharmacological toxicity. The toxicity profile of the active PROTAC MZ 1 has not been fully described. As a research chemical, cis-MZ 1 hydrate should be handled with standard laboratory safety precautions. No specific toxicological data is available for this compound. For research use only, not for human consumption.
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| References | |
| Additional Infomation |
The Bromo- and Extra-Terminal (BET) proteins BRD2, BRD3, and BRD4 are key regulators of transcription, epigenetics, and cancer, and serve as the targets of the pan-BET selective bromodomain inhibitor JQ1. However, the lack of selectivity among BET family members limits the utility of current inhibitors for target validation and may lead to off-target effects or toxicity in therapeutic applications. To address this, we designed Proteolysis-Targeting Chimeras (PROTACs) that link JQ1 to a ligand for the E3 ubiquitin ligase VHL, aiming to induce targeted degradation of BET proteins. One such compound, MZ1, potently and rapidly induces reversible, long-lasting, and unexpectedly selective degradation of BRD4 over BRD2 and BRD3. This activity is dependent on VHL binding and occurs at concentrations low enough to avoid stabilization of HIF-1α. Analysis of gene expression profiles of cancer-related genes responsive to JQ1 reveals that MZ1 induces a distinct and more restricted transcriptional response, consistent with selective suppression of BRD4. These findings open new avenues for exploring the cellular functions and therapeutic potential of selective BRD4 targeting. [1]
cis-MZ 1 hydrate is the cis-isomer of MZ 1, a first-generation BRD4-targeting PROTAC. The active PROTAC MZ 1 (compound 1 in the original publication) degrades BRD4 with DC50 values in the low nanomolar range. cis-MZ 1 is a valuable negative control for PROTAC experiments to rule out non-degradative effects. For research use only. MZ 1 was first described by Zengerle et al. (2015) in the Journal of Medicinal Chemistry. |
| Molecular Formula |
C49H60CLN9O8S2.1/4H2O
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|---|---|
| Molecular Weight |
1007.15
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| Appearance |
White to off-white 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) |
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
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.9929 mL | 4.9645 mL | 9.9290 mL | |
| 5 mM | 0.1986 mL | 0.9929 mL | 1.9858 mL | |
| 10 mM | 0.0993 mL | 0.4965 mL | 0.9929 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.
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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