| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 500mg |
| Targets |
The (S,R,S)-AHPC-PEG3-NH2 hydrochloride acts as the core structural unit of the VHL (Von Hippel-Lindau) ligand in the BET (Bromodomain and Extra-Terminal) degrading PROTACs (Proteolysis-Targeting Chimeras) studied. The PROTACs containing this compound simultaneously target two types of proteins:
1. VHL tumor suppressor protein (the target of the VHL ligand moiety where (S,R,S)-AHPC-PEG3-NH2 hydrochloride is located); 2. BET family proteins (e.g., Brd4, Brd2), the targets of the BET inhibitor moiety (JQ1 or I-BET726) in PROTACs. Isothermal titration calorimetry (ITC) data for the PROTACs (containing this VHL ligand unit) showed that the VCB complex (VHL-Elongin C-Elongin B) bound to the PROTACs alone with a Kd of 110 nM, while the Kd values for VCB binding to the binary complexes of PROTACs with Brd4 BD2 (second bromodomain of Brd4) and Brd2 BD1 (first bromodomain of Brd2) were 180 nM and 330 nM, respectively [1] |
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| ln Vitro |
All in vitro activity data were obtained from PROTACs containing (S,R,S)-AHPC-PEG3-NH2 hydrochloride (as the VHL ligand moiety), rather than the compound alone:
1. BET protein degradation in HeLa cells: HeLa cells were treated with the PROTACs for 24 hours. After treatment, protein levels of BET family members (e.g., Brd4, Brd2) were detected by immunoblotting and quantified relative to the DMSO control group. Triazolodiazepine (JQ1)-derived PROTACs (containing this VHL ligand unit) exhibited more potent BET degradation activity than tetrahydroquinoline (I-BET726)-derived PROTACs [1] 2. Antiproliferative activity in MV4;11 acute myeloid leukemia (AML) cells: MV4;11 cells were treated with the PROTACs or their parent BET ligands (JQ1/I-BET726) for 48 hours, followed by cell viability quantification. The antiproliferative activity of the PROTACs was driven by cMyc and showed a marked dependence on the length of the polyethylene glycol (PEG) linker (connecting the BET inhibitor and (S,R,S)-AHPC-PEG3-NH2 hydrochloride) [1] 3. cMyc suppression in MV4;11 cells: MV4;11 cells were treated with 50 nM of the PROTACs, parent BET inhibitors, or DMSO for 4 hours. cMyc protein levels were detected by immunoblotting, and the PROTACs showed significant cMyc suppression effects compared to the control group [1] |
| Enzyme Assay |
Isothermal Titration Calorimetry (ITC) was used to measure the cooperativity of ternary complex formation between the PROTACs (containing (S,R,S)-AHPC-PEG3-NH2 hydrochloride), VCB complex (VHL-Elongin C-Elongin B), and BET proteins. The detailed experimental process included three groups:
1. Titrate the VCB complex into a solution containing only the PROTACs (to determine the binding affinity between VCB and the PROTACs); 2. Titrate the VCB complex into a binary complex solution composed of Brd4 BD2 (the second bromodomain of Brd4) and the PROTACs (to evaluate the binding affinity between VCB and the Brd4 BD2-PROTAC binary complex); 3. Titrate the VCB complex into a binary complex solution composed of Brd2 BD1 (the first bromodomain of Brd2) and the PROTACs (to evaluate the binding affinity between VCB and the Brd2 BD1-PROTAC binary complex). Binding affinity data (Kd values) were calculated from the ITC results: the Kd for VCB binding to the PROTACs alone was 110 nM, while the Kd values for VCB binding to the Brd4 BD2-PROTAC and Brd2 BD1-PROTAC binary complexes were 180 nM and 330 nM, respectively. These results indicated negative cooperativity in the ternary complex formation of tetrahydroquinoline (I-BET726)-derived PROTACs [1] |
| Cell Assay |
Two main cell-based assays were conducted using PROTACs containing (S,R,S)-AHPC-PEG3-NH2 hydrochloride, with no separate assays for the compound alone:
1. BET protein degradation assay in HeLa cells: HeLa cells were seeded and cultured to the appropriate density, then treated with different concentrations of the PROTACs for 24 hours. After the treatment period, the cells were lysed to extract total proteins, and the protein levels of BET family members (e.g., Brd4, Brd2) were detected by immunoblotting. The intensity of the protein bands was quantified using relevant software, with the DMSO-treated group as the reference (set to 100% protein level). The experiment included two biological replicates, and one representative set of results was presented in the study [1] 2. Antiproliferative and cMyc suppression assay in MV4;11 cells: - For antiproliferative activity: MV4;11 cells (AML cell line) were seeded in a suitable culture plate and treated with serial concentrations of the PROTACs or their parent BET ligands (JQ1/I-BET726) for 48 hours. After incubation, cell viability was measured using a cell viability assay method, and the half-effective concentrations (EC50) of the PROTACs and parent ligands were calculated based on the viability data; - For cMyc suppression: MV4;11 cells were treated with 50 nM of the PROTACs, parent BET inhibitors, or DMSO (control) for 4 hours. The cells were then lysed to extract proteins, and cMyc protein levels were detected by immunoblotting. Similar to the HeLa cell assay, two biological replicates were performed, and one representative result was shown [1] |
| References | |
| Additional Infomation |
1. Structural role in PROTACs: (S,R,S)-AHPC-PEG3-NH2 hydrochloride is the core structural component of the VHL ligand in the bifunctional BET degradation PROTACs designed in this study. These PROTACs consist of two functional parts—a BET bromine domain inhibitor (JQ1 or I-BET726) and a VHL ligand (containing (S,R,S)-AHPC-PEG3-NH2 hydrochloride)—linked by a polyethylene glycol (PEG) linker of a specific length [1]. 2. Key findings related to linkers and synergistic effects: Studies have shown that a more effective BET inhibitor (I-BET726) does not necessarily produce more effective PROTACs. Specifically, tetrahydroquinoline (I-BET726)-derived PROTACs (containing the VHL ligand unit) exhibited negative synergism in the formation of ternary complexes (with VCB and BET proteins) and weak BET degradation and antiproliferative activity, while triazolidinedazapine (JQ1)-derived PROTACs exhibited positive synergism and stronger biological activity. These results highlight the crucial role of conjugation strategies (including the length of the PEG linker connecting the BET inhibitor and (S,R,S)-AHPC-PEG3-NH2 hydrochloride and the exit vector of the inhibitor) in determining the efficacy of PROTACs [1]. 3. Significance of the study: The insights and structure-activity relationship (SAR) framework of divalent degraders (including degraders containing (S,R,S)-AHPC-PEG3-NH2 hydrochloride) provided in this study are expected to be widely used in the design and optimization of PROTACs in the future [1].
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| Molecular Formula |
C30H46CLN5O7S
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|---|---|
| Molecular Weight |
656.233545780182
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| Exact Mass |
655.28
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| CAS # |
2097971-11-2
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| PubChem CID |
131704494
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| Appearance |
Light yellow to brown solid powder
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
17
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| Heavy Atom Count |
44
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| Complexity |
884
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| Defined Atom Stereocenter Count |
3
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| SMILES |
C(N1C[C@H](O)C[C@H]1C(=O)NCC1C=CC(C2SC=NC=2C)=CC=1)(=O)[C@H](C(C)(C)C)NC(=O)COCCOCCOCCN.Cl
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| InChi Key |
ZOYHUTRHKHRRPK-QVRKWNSCSA-N
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| InChi Code |
InChI=1S/C30H45N5O7S.ClH/c1-20-26(43-19-33-20)22-7-5-21(6-8-22)16-32-28(38)24-15-23(36)17-35(24)29(39)27(30(2,3)4)34-25(37)18-42-14-13-41-12-11-40-10-9-31;/h5-8,19,23-24,27,36H,9-18,31H2,1-4H3,(H,32,38)(H,34,37);1H/t23-,24+,27-;/m1./s1
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| Chemical Name |
(2S,4R)-1-[(2S)-2-[[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide;hydrochloride
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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: (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)
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| Solubility (In Vitro) |
DMSO : ~200 mg/mL (~304.77 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (76.19 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5239 mL | 7.6193 mL | 15.2386 mL | |
| 5 mM | 0.3048 mL | 1.5239 mL | 3.0477 mL | |
| 10 mM | 0.1524 mL | 0.7619 mL | 1.5239 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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