TMX1
Based on 1 Customer Validation
TMX1 is a covalent, selective BRD4 molecular glue degrader. TMX1 binds to the JQ1-binding site of BRD4BD2, forms covalent bonds with Cys58 of DCAF16 and Cys87 of GAK in a BRD4BD2-dependent template-assisted manner, stabilizes the BRD4-TMX1-DCAF16 ternary complex, and promotes the ubiquitination of BRD4 via the CRL4DCAF16 ubiquitin ligase complex. TMX1 induces selective degradation of BRD4, mild degradation of BRD2 and BRD3, as well as DCAF16-dependent cytotoxicity.
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- Pureté : 99.57%
- CAS No.: 3079093-92-5
- Formule: C26H28N4O3S
- Masse moléculaire:476.59
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Stockage:
4°C, sealed storage, away from moisture and light
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)
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Activité biologique
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Chemical Information
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Solvant et solubilité
- Protocole
- Pureté et documentation
- Références
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Help & FAQs
Activité biologique
Description
IC50 & Target
[1]|
BRD4 BD2 |
BRD2 |
BRD3 |
In Vitro
TMX1 (0.01-10 μM; 4-24 h) induces proteasome-dependent, DCAF16-mediated degradation of BRD4, with selective activity against the BRD4BD2 domain, achieving ~80% maximal degradation in K562 cells at 16 h[1].
TMX1 (0.5 μM; 5 h) selectively downregulates BRD4 (with minor effects on BRD2 and BRD3) in K562 cells[1].
TMX1 (0.1 μM; 14 days) requires DCAF16 for its induced cellular toxicity in K562 cells[1].
TMX1 (0.01-10 μM; 16 h) induces concentration-dependent degradation of BRD4 in K562 cells after 16 h of treatment[2].
TMX1 (1 μM; 16 h) induced degradation of BRD4_BD2-eGFP in K562-Cas9 cells requires the RBX1-CUL4-DDB1-DCAF16 ubiquitin ligase complex, as demonstrated by a UPS-focused CRISPR screen with treatment at 1 μM for 16 h[2].
TMX1 (1 μM; 16 h) induced BRD4 degradation in K562 cells requires DCAF16 after 16 h of treatment at 1 μM[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:K562 cells
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Concentration:0.01-10 μM
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Incubation Time:16 h
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Result:Induced concentration-dependent degradation of BRD4, with visible reduction of BRD4 protein levels across all tested concentrations relative to the DMSO control.
Maintained stable actin levels as a loading control.
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Cell Line:DCAF16-knockout K562 cells
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Concentration:1 μM
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Incubation Time:16 h
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Result:Abolished TMX1-induced BRD4 degradation in DCAF16-knockout K562 cells, whereas BRD4 degradation was observed in cells transduced with non-targeting sgRNA.
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Cell Line:DCAF16-knockout K562 cells expressing DCAF16 mutants
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Concentration:1 μM
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Incubation Time:16 h
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Result:Abolished TMX1-induced BRD4 degradation in cells expressing the DCAF16 C58S mutant, whereas degradation was preserved in cells expressing wild-type DCAF16 or other mutants (C177S, C179S, A53R).
Chemical Information
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CAS No. 3079093-92-5
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Appearance Solid
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Masse moléculaire 476.59
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Formule C26H28N4O3S
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Color White to off-white
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SMILES
O=C/C=C/C1=CC=C(C2=N[C@@H](CC(OC(C)(C)C)=O)C3=NN=C(C)N3C4=C2C(C)=C(C)S4)C=C1
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Livraison
Room temperature in continental US; may vary elsewhere.
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Stockage
4°C, sealed storage, away from moisture and light
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)
Solvant et solubilité
In Vitro:
DMSO : ≥ 100 mg/mL (209.82 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
* "≥" means soluble, but saturation unknown.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (5.25 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (5.25 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL. * In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocole
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Mammalian live/dead viability and cytotoxicity staining
Live/dead viability and cytotoxicity staining assays are based on the simultaneous detection of intracellular esterase activity in metabolically active (viable) cells and membrane integrity loss in non-viable cells. In commonly used dual-staining approaches, membrane-permeant fluorogenic substrates are converted by intracellular esterases into fluorescent products in live cells, while impermeant DNA-binding dyes selectively enter cells with compromised plasma membranes and label nucleic acids in dead or dying cells, enabling discrimination between viable and non-viable populations by fluorescence microscopy or flow cytometry.
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Cell Viability Determination by MTT Colorimetric Assay
The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.
Pureté et documentation
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Fiche technique (280 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Instruction de manipulation (2659 KB)
Références
[1]. Li YD, et al. Template-assisted covalent modification of DCAF16 underlies activity of BRD4 molecular glue degraders. bioRxiv [Preprint]. 2023 Feb 15:2023.02.14.528208. [Content Brief]
[2]. Li YD, et al. Template-assisted covalent modification underlies activity of covalent molecular glues. Nat Chem Biol. 2024;20(12):1640-1649. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 2.0982 mL | 10.4912 mL | 20.9824 mL | 52.4560 mL |
| 5 mM | 0.4196 mL | 2.0982 mL | 4.1965 mL | 10.4912 mL | |
| 10 mM | 0.2098 mL | 1.0491 mL | 2.0982 mL | 5.2456 mL | |
| 15 mM | 0.1399 mL | 0.6994 mL | 1.3988 mL | 3.4971 mL | |
| 20 mM | 0.1049 mL | 0.5246 mL | 1.0491 mL | 2.6228 mL | |
| 25 mM | 0.0839 mL | 0.4196 mL | 0.8393 mL | 2.0982 mL | |
| 30 mM | 0.0699 mL | 0.3497 mL | 0.6994 mL | 1.7485 mL | |
| 40 mM | 0.0525 mL | 0.2623 mL | 0.5246 mL | 1.3114 mL | |
| 50 mM | 0.0420 mL | 0.2098 mL | 0.4196 mL | 1.0491 mL | |
| 60 mM | 0.0350 mL | 0.1749 mL | 0.3497 mL | 0.8743 mL | |
| 80 mM | 0.0262 mL | 0.1311 mL | 0.2623 mL | 0.6557 mL | |
| 100 mM | 0.0210 mL | 0.1049 mL | 0.2098 mL | 0.5246 mL |
TMX1 Related Classifications
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