| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
Purity: ≥98%
AT6 is a novel, potent and highly selective PROTAC-based BRD degrader, it is analog of AT1. Inducing macromolecular interactions with small molecules to activate cellular signaling is a challenging goal. PROTACs (proteolysis-targeting chimeras) are bifunctional molecules that recruit a target protein in proximity to an E3 ubiquitin ligase to trigger protein degradation. Structural elucidation of the key ternary ligase-PROTAC-target species and its impact on target degradation selectivity remain elusive.
| Targets |
AT6 targets bromodomain and extra-terminal (BET) family protein BRD4 (bromodomain-containing protein 4, BD2 domain) and von Hippel-Lindau (VHL) E3 ubiquitin ligase. [1]
|
|---|---|
| ln Vitro |
In order to cause protein degradation, bifunctional entities known as PROTACs (proteolysis-targeting chimaeras) enlist a target protein that is close to an E3 ubiquitin ligase. It is yet unclear how the target species' structural makeup affects the selectivity of target degradation by the important ternary ligase PROTAC. In the ternary complex, the ligand folds inward to permit the development of certain intermolecular interactions. Studies using isothermal titration calorimetry are compatible with the pronounced cooperative production of ternary complexes with Brd4BD2, which is corroborated by surface mutagenesis and proximity experiments. Compound AT1, which was created based on structure, shows extremely selective Brd4 depletion in cells[1].
1. AT6 mediates selective cooperative recognition of BRD4 BD2 and VHL, forming a stable ternary complex. Biochemical assays confirm that AT6 exhibits preferential binding to BRD4 BD2 over BRD4 BD1 and non-BET bromodomains, with no significant cross-reactivity. [1] 2. AT6 induces conformational changes in both BRD4 BD2 and VHL to facilitate mutual interaction: The BRD4-binding moiety of AT6 interacts with BRD4 BD2 via hydrophobic and hydrogen-bonding networks, while its VHL-binding ligand triggers a structural shift in VHL, creating a complementary hydrophobic pocket for BRD4 BD2. This cooperative binding enhances the stability of the BRD4-AT6-VHL ternary complex. [1] 3. AT6 demonstrates efficient ternary complex formation capacity, as validated by biochemical and structural analyses, supporting its potential to induce BRD4 degradation via the VHL-dependent ubiquitin-proteasome pathway . [1] |
| Enzyme Assay |
1. Ternary complex formation assay (AlphaScreen): Recombinant BRD4 BD2 domain and VHL-HIF1α fusion protein are mixed with serial dilutions of AT6. The assay measures the proximity between fluorophore-labeled BRD4 and VHL, with signal amplification indicating ternary complex formation. Experiments are performed in triplicate, and data are normalized to a positive control (MZ1) to assess relative binding efficiency. [1]
2. Surface Plasmon Resonance (SPR) assay: Recombinant BRD4 BD2 or BD1 domain is immobilized on a sensor chip. AT6 is serially diluted in running buffer and injected over the chip to monitor binding kinetics (association and dissociation rates) and affinity. A parallel assay with recombinant VHL-HIF1α complex is conducted to confirm direct binding of AT6 to VHL. [1] 3. Isothermal Titration Calorimetry (ITC) assay: AT6 is titrated into solutions containing recombinant BRD4 BD2 or VHL-HIF1α complex at 25°C. Heat changes associated with binding are recorded to determine thermodynamic parameters (ΔH, ΔS, and binding stoichiometry), which reflect the cooperative nature of the interaction between AT6 and its two targets. Experiments are performed in duplicate to ensure reproducibility. [1] |
| References | |
| Additional Infomation |
1. AT6 is a protein hydrolysis-targeting chimera (PROTAC) designed as a tool compound for studying the structural basis of PROTAC-mediated co-recognition. Its structure comprises a BRD4 BD2 selective binding moiety, a rigid linker, and a VHL binding ligand (derived from VH032). [1] 2. AT6 differs from other BET PROTACs (such as MZ1) in its linker design and BRD4 binding specificity, providing a comparative model for studying how structural modifications affect co-binding and target selectivity. [1] 3. Structural analysis of the BRD4-AT6-VHL ternary complex revealed key molecular interactions driving co-recognition, providing insights for the rational design of next-generation BET degraders with higher potency and tissue selectivity. [1]
|
| Molecular Formula |
C48H58CLN9O7S3
|
|---|---|
| Molecular Weight |
1004.67822599411
|
| Exact Mass |
1003.33
|
| CAS # |
2098836-50-9
|
| PubChem CID |
124201839
|
| Appearance |
White to off-white solid powder
|
| LogP |
4.6
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
14
|
| Rotatable Bond Count |
20
|
| Heavy Atom Count |
68
|
| Complexity |
1740
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
CC1=C(SC2N3C(=NN=C3[C@H](CC(=O)NCCOCCOCCSC(C)(C)[C@H](NC(=O)C)C(N3C[C@H](O)C[C@H]3C(=O)NCC3C=CC(C4SC=NC=4C)=CC=3)=O)N=C(C3C=CC(Cl)=CC=3)C1=2)C)C
|
| InChi Key |
ZCEIHPCVOJGWHG-TZPPCSJFSA-N
|
| InChi Code |
InChI=1S/C48H58ClN9O7S3/c1-27-29(3)68-47-40(27)41(33-12-14-35(49)15-13-33)54-37(44-56-55-30(4)58(44)47)23-39(61)50-16-17-64-18-19-65-20-21-67-48(6,7)43(53-31(5)59)46(63)57-25-36(60)22-38(57)45(62)51-24-32-8-10-34(11-9-32)42-28(2)52-26-66-42/h8-15,26,36-38,43,60H,16-25H2,1-7H3,(H,50,61)(H,51,62)(H,53,59)/t36-,37+,38+,43-/m1/s1
|
| Chemical Name |
(2S,4R)-1-[(2R)-2-acetamido-3-[2-[2-[2-[[2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetrazatricyclo[8.3.0.02,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetyl]amino]ethoxy]ethoxy]ethylsulfanyl]-3-methylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide
|
| Synonyms |
AT6; AT-6; AT 6
|
| 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: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), 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)
|
| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~99.53 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (4.98 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 5 mg/mL (4.98 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 5 mg/mL (4.98 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.9953 mL | 4.9767 mL | 9.9534 mL | |
| 5 mM | 0.1991 mL | 0.9953 mL | 1.9907 mL | |
| 10 mM | 0.0995 mL | 0.4977 mL | 0.9953 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.
 The crystal structure of the Brd4BD2:MZ1:VHL-ElonginC-ElonginB complex.Nat Chem Biol.2017 May;13(5):514-521. |
|---|
 Brd4BD2and VHL form a stable, cooperative complex in the presence of MZ1.
Schematic model of selective PROTAC-induced target degradation.Nat Chem Biol.2017 May;13(5):514-521. |
 The molecular basis of MZ1-induced compact complex formation between Brd4BD2and VHL.
Structure-guided design and characterization of Brd4-selective degrader AT1.Nat Chem Biol.2017 May;13(5):514-521. |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
