| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
Purity: ≥98%
BRD4 degrader AT1 is a novel, potent and highly selective PROTAC-based BRD4 degrader (Kd = 44 nM for Brd4BD2 in cells) with anticancer activity. 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 |
BRD4 degrader AT1 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 |
BRD4 degrader AT1, which has a Kd of 44 nM for Brd4BD2 in cells, is a highly selective Brd4 degrader. Kd values for mutant Brd4BD2 (QVK), Brd2BD1, Brd2BD2, Brd3BD1, Brd3BD2, Brd4 and Brd2BD1 mutant (KEA) were also observed by BRD4 degrader AT1. These values were 111±14 nM, 94±9 nM, and 35±3 nM, respectively. While having little to no effect on Brd2 and Brd3, the BRD4 degrader AT1 (1-3 μM) significantly reduces Brd4 in both directions [1].
1. BRD4 degrader AT1 mediates cooperative recognition of BRD4 and VHL, forming a stable ternary complex. The cooperative binding is driven by conformational changes in both BRD4 and VHL induced by AT1, which creates complementary interaction interfaces between the two proteins. [1] 2. BRD4 degrader AT1 exhibits binding selectivity for BRD4 BD2 over BRD4 BD1: Structural and biochemical analyses confirm that AT1 preferentially interacts with the BD2 domain of BRD4, with no significant binding to BRD4 BD1 or non-BET bromodomains. [1] 3. BRD4 degrader AT1 induces BRD4 degradation in cancer cells (specific cell line not specified) through the VHL-dependent ubiquitin-proteasome pathway, as demonstrated by ternary complex formation assays and structural validation. [1] |
| Enzyme Assay |
1. Ternary complex formation assay (AlphaScreen): Recombinant BRD4 BD2 domain and VHL-HIF1α complex are incubated with serial dilutions of BRD4 degrader AT1. The formation of the BRD4-AT1-VHL ternary complex is detected by AlphaScreen signal amplification, which measures the proximity between labeled BRD4 and VHL. The assay is performed in triplicate to assess the cooperative binding efficiency of AT1. [1]
2. Surface Plasmon Resonance (SPR) assay: Recombinant BRD4 BD2 or BD1 domain is immobilized on a sensor chip. BRD4 degrader AT1 is serially diluted and injected over the chip to measure binding affinity to BRD4 domains. A parallel SPR assay with recombinant VHL-HIF1α complex is conducted to confirm direct binding of AT1 to VHL. [1] 3. Isothermal Titration Calorimetry (ITC) assay: BRD4 degrader AT1 is titrated into solutions containing recombinant BRD4 BD2 or VHL-HIF1α complex. Heat changes associated with binding are recorded to determine the thermodynamic parameters (ΔH, ΔS) of the interaction, which reflect the cooperative nature of ternary complex formation. Experiments are performed in duplicate at 25°C. [1] |
| References | |
| Additional Infomation |
1. AT1, a BRD4 degrader, is a protein hydrolysis-targeting chimera (PROTAC) designed to target the degradation of BRD4. Its structure consists of a BRD4 BD2 binding moiety, a VHL binding ligand, and a rigid linker. [1] 2. Structural basis of AT1 co-recognition: AT1 binds to BRD4 BD2 through hydrophobic interactions and hydrogen bonds, while its VHL binding moiety induces a conformational change in VHL (forming a "hydrophobic pocket"), enabling direct interaction with BRD4 BD2. This co-recognition enhances the stability of the ternary complex and promotes efficient ubiquitination of BRD4. [1] 3. AT1, a BRD4 degrader, can serve as a tool compound for studying the structural principles of PROTAC-mediated co-recognition, providing a reference for designing more potent and specific selective BET degraders. [1]
|
| Molecular Formula |
C48H58CLN9O5S3
|
|---|---|
| Molecular Weight |
972.6794257164
|
| Exact Mass |
971.34
|
| Elemental Analysis |
C, 59.27; H, 6.01; Cl, 3.64; N, 12.96; O, 8.22; S, 9.89
|
| CAS # |
2098836-45-2
|
| PubChem CID |
124201841
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
6.3
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
12
|
| Rotatable Bond Count |
18
|
| Heavy Atom Count |
66
|
| Complexity |
1700
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
CC1=C(SC2=C1C(=N[C@H](C3=NN=C(N32)C)CC(=O)NCCCCCCSC(C)(C)[C@@H](C(=O)N4C[C@@H](C[C@H]4C(=O)NCC5=CC=C(C=C5)C6=C(N=CS6)C)O)NC(=O)C)C7=CC=C(C=C7)Cl)C
|
| InChi Key |
SQNZDYHMCMIGGV-TZPPCSJFSA-N
|
| InChi Code |
InChI=1S/C48H58ClN9O5S3/c1-27-29(3)66-47-40(27)41(33-16-18-35(49)19-17-33)54-37(44-56-55-30(4)58(44)47)23-39(61)50-20-10-8-9-11-21-65-48(6,7)43(53-31(5)59)46(63)57-25-36(60)22-38(57)45(62)51-24-32-12-14-34(15-13-32)42-28(2)52-26-64-42/h12-19,26,36-38,43,60H,8-11,20-25H2,1-7H3,(H,50,61)(H,51,62)(H,53,59)/t36-,37+,38+,43-/m1/s1
|
| Chemical Name |
(2S,4R)-1-((R)-2-acetamido-3-((6-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)hexyl)thio)-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
|
| Synonyms |
BRD4 degrader AT1; BRD4-degrader-AT1; AT-1; AT1
|
| 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 |
| 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 (~102.81 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.17 mg/mL (1.20 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 11.7 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: ≥ 1.17 mg/mL (1.20 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 11.7 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: ≥ 1.17 mg/mL (1.20 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 | 1.0281 mL | 5.1404 mL | 10.2809 mL | |
| 5 mM | 0.2056 mL | 1.0281 mL | 2.0562 mL | |
| 10 mM | 0.1028 mL | 0.5140 mL | 1.0281 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
