| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
SGK3 and VPS34 (DC50 = 3~10 nM)
HaloPROTAC-E targets HaloTag-fused proteins of interest through the HaloTag ligand moiety, while the E3 ligase recruiting moiety recruits an E3 ubiquitin ligase (typically CRBN or VHL). This brings the target protein into proximity with the ubiquitin-proteasome machinery, leading to ubiquitination and subsequent proteasomal degradation of the target protein. The compound's mechanism is based on the PROTAC technology, which induces targeted protein degradation rather than simple inhibition. |
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| ln Vitro |
In vitro studies have demonstrated that HaloPROTAC-E effectively induces degradation of HaloTag-fused proteins in cellular systems. The compound's activity is assessed by measuring the reduction in target protein levels using Western blot analysis or other protein detection methods. The efficiency of degradation depends on the concentration of the compound, incubation time, and the specific target protein. HaloPROTAC-E has been shown to induce rapid and potent degradation of HaloTag fusion proteins.
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| ln Vivo |
In vivo studies for HaloPROTAC-E have been limited, as the compound is primarily used as a research tool in cell-based studies. The compound's potential for in vivo applications depends on its pharmacokinetic properties, including oral bioavailability and tissue distribution. Studies in appropriate animal models would be needed to evaluate its in vivo efficacy, pharmacokinetics, and safety profile. The compound is primarily used for chemical biology research rather than therapeutic development.
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| Enzyme Assay |
For protein degradation assays, cells expressing HaloTag-fused proteins of interest are treated with HaloPROTAC-E at various concentrations (typically 0.001-10 μM) for 4-24 hours. Target protein levels are assessed by Western blot using antibodies against the protein of interest or against the HaloTag. Degradation kinetics are evaluated by time-course experiments. The compound's effect on cell viability is assessed using MTT or CellTiter-Glo assays to ensure specificity of degradation.
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| Cell Assay |
For cellular studies, appropriate cell lines expressing HaloTag-fused target proteins are cultured in appropriate media and treated with HaloPROTAC-E at various concentrations for 4-24 hours. Target protein degradation is assessed by Western blot using antibodies against the protein of interest or the HaloTag. Ubiquitination of the target protein is assessed by immunoprecipitation followed by Western blot using anti-ubiquitin antibodies. Cellular phenotypes resulting from target protein degradation are evaluated using appropriate functional assays.
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| Animal Protocol |
For in vivo studies, appropriate animal models would be used to evaluate HaloPROTAC-E. The compound would be administered via intravenous, intraperitoneal, or oral routes at various doses. Target protein degradation in tissues would be assessed by Western blot or immunohistochemistry. Pharmacokinetic parameters including half-life, clearance, and tissue distribution would be determined. However, detailed in vivo protocols for HaloPROTAC-E have not been extensively reported.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of HaloPROTAC-E have not been fully characterized in the available literature. As a PROTAC molecule, it is expected to have higher molecular weight and potentially lower oral bioavailability compared to traditional small molecules. The compound is typically handled as a solid and stored at -20°C to maintain stability. Comprehensive PK studies are needed to fully understand its ADME properties.
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| Toxicity/Toxicokinetics |
HaloPROTAC-E is a research compound intended for laboratory use only and is not approved for human therapeutic use. As a PROTAC molecule, it induces degradation of HaloTag-fused proteins through the ubiquitin-proteasome system. Standard laboratory safety precautions should be followed when handling this compound. Storage at -20°C is recommended.
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| References | |
| Additional Infomation |
Inducing post-translational modifications of proteins is an important method for studying biology and validating drug targets. An effective strategy is to express target proteins fused with exogenous tags, thereby utilizing tag-directed chemical degraders to mediate protein ubiquitination and proteasome degradation. In this paper, we combine an improved HaloPROTAC degradation probe with CRISPR/Cas9 gene editing technology to trigger the rapid degradation of endogenous target proteins. Our optimized probe, HaloPROTAC-E, is a chloroalkane conjugate of the high-affinity VHL-binding protein VH298, capable of inducing the reversible degradation of two endosomal localization proteins, SGK3 and VPS34, with DC50 values of 3–10 nM. HaloPROTAC-E rapidly (approximately 50% degradation after 30 minutes) and completely (maximum degradation rate of approximately 95% after 48 hours) consumes Halo-tagged SGK3, thereby blocking downstream phosphorylation of the SGK3 substrate NDRG1. Compared with previously reported HaloPROTAC3 compounds, HaloPROTAC-E can more effectively induce the steady-state degradation of Halo-tagged endogenous VPS34. Quantitative proteomics analysis showed that HaloPROTAC-E has significant selectivity, inducing only the degradation of Halo-tagged endogenous VPS34 complexes (VPS34, VPS15, Beclin1, and ATG14), while the degradation of other proteins was not significant. This study shows that combining HaloPROTAC with CRISPR/Cas9 endogenous protein labeling technology is an effective method for inducing rapid and reversible degradation of endogenous proteins, which can be used to study their functions. [1]
HaloPROTAC-E is a PROTAC molecule that induces degradation of HaloTag-fused proteins through the ubiquitin-proteasome system. It contains a HaloTag ligand and an E3 ubiquitin ligase recruiting moiety. The compound is used as a research tool for chemical biology and drug discovery. It is for research use only and has not been approved for clinical applications. Storage at -20°C is recommended. |
| Molecular Formula |
C39H56CLN5O8S
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|---|---|
| Molecular Weight |
790.408648490906
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| Exact Mass |
789.353
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| CAS # |
2365478-58-4
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| PubChem CID |
154723985
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| Appearance |
Colorless to light yellow ointment
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| LogP |
4.1
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
24
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| Heavy Atom Count |
54
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| Complexity |
1250
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| Defined Atom Stereocenter Count |
3
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| SMILES |
CC1=C(SC=N1)C2=CC(=C(C=C2)CNC(=O)[C@@H]3C[C@H](CN3C(=O)[C@H](C(C)(C)C)NC(=O)C4(CC4)C#N)O)OCCOCCOCCOCCCCCCCl
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| InChi Key |
NJCGXLMTZGCSRS-JSWXEYCZSA-N
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| InChi Code |
InChI=1S/C39H56ClN5O8S/c1-27-33(54-26-43-27)28-9-10-29(32(21-28)53-20-19-52-18-17-51-16-15-50-14-8-6-5-7-13-40)23-42-35(47)31-22-30(46)24-45(31)36(48)34(38(2,3)4)44-37(49)39(25-41)11-12-39/h9-10,21,26,30-31,34,46H,5-8,11-20,22-24H2,1-4H3,(H,42,47)(H,44,49)/t30-,31+,34-/m1/s1
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| Chemical Name |
(2S,4R)-N-[[2-[2-[2-[2-(6-chlorohexoxy)ethoxy]ethoxy]ethoxy]-4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1-[(2S)-2-[(1-cyanocyclopropanecarbonyl)amino]-3,3-dimethylbutanoyl]-4-hydroxypyrrolidine-2-carboxamide
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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 |
| 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 : 100 mg/mL (126.52 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.16 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 25.0 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: ≥ 2.5 mg/mL (3.16 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2652 mL | 6.3258 mL | 12.6517 mL | |
| 5 mM | 0.2530 mL | 1.2652 mL | 2.5303 mL | |
| 10 mM | 0.1265 mL | 0.6326 mL | 1.2652 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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