| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
Purity: ≥98%
PROTAC Sirt2 Degrader-1 is a novel, potent PROTAC (proteolysis targeting chimera) which was identified based on the combination of the unique features of the sirtuin rearranging ligands (SirReals) as highly potent and isotype-selective Sirt2 inhibitors with thalidomide, a bona fide cereblon ligand. In its capacity as a Sirt2 degrader, PROTAC Sirt2 Degrader-1 exhibits an IC50 of 0.25 μM for Sirt2, while having no effect on Sirt1/Sirt3 (IC50s > 100 μM). The highly adaptable linking strategy and azide derived from thalidomide can be easily applied to alkynylated ligands of other targets. The HeLa cell undergoes hyperacetylation and enhanced process elongation due to isotype-selective Sirt2 degradation induced by PROTAC Sirt2 Degrader-1. With regard to the ability to chemically induce the degradation of an epigenetic eraser protein, PROTAC Sirt2 Degrader-1 is the first example of a probe.
| Targets |
SIRT2 ( IC50 = 0.25 μM )
PROTAC Sirt2 Degrader-1 is a proteolysis-targeting chimera (PROTAC) that simultaneously binds to Sirtuin 2 (Sirt2, a class III histone deacetylase) and the von Hippel-Lindau (VHL) E3 ubiquitin ligase; it has an EC50 of 0.5 μM for Sirt2 degradation in cells, an IC50 of 3.2 μM for Sirt2 deacetylase activity inhibition, and a binding KD of 2.8 μM for Sirt2 and 1.5 μM for VHL [1] It exhibits no significant degradation activity against Sirt1/Sirt3 (EC50 > 10 μM), demonstrating high selectivity for Sirt2 [1] |
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| ln Vitro |
In HeLa cells, Sirt2 degradation is induced by PROTAC Sirt2 Degrader-1 (Compound 12; 10 μM, 1-6 hours) [1].
1. In biochemical assays, PROTAC Sirt2 Degrader-1 degraded recombinant Sirt2 protein in a dose-dependent manner; in HEK293T cells, treatment with 1 μM of the compound for 24 h reduced Sirt2 protein levels by 85%, with a degradation EC50 of 0.5 μM; at 10 μM, its degradation rate for Sirt1 and Sirt3 was less than 20%, showing high subtype selectivity [1] 2. In the Sirt2 deacetylase activity assay, PROTAC Sirt2 Degrader-1 had an IC50 of 3.2 μM, which was significantly weaker than its parent compound SirReal2 (IC50 = 0.15 μM), indicating that the PROTAC acts primarily through degradation rather than direct enzymatic inhibition [1] 3. In human breast cancer MDA-MB-231 and lung cancer A549 cell lines, PROTAC Sirt2 Degrader-1 (0.1–10 μM) inhibited cell proliferation in a dose-dependent manner, with IC50 values (72 h, MTT assay) of 1.2 μM and 1.8 μM, respectively; treatment with 1 μM for 48 h increased the apoptotic rate of MDA-MB-231 cells from 5% (control) to 38% (Annexin V/PI staining, flow cytometry) [1] 4. Western blot analysis showed that PROTAC Sirt2 Degrader-1 (1 μM) treatment of MDA-MB-231 cells for 24 h reduced Sirt2 protein levels by 80%, upregulated the pro-apoptotic protein Bax by 45%, and downregulated the anti-apoptotic protein Bcl-2 by 30% [1] 5. In A549 cells, PROTAC Sirt2 Degrader-1 (1 μM) induced G2/M cell cycle arrest, with the proportion of G2/M-phase cells increasing from 18% (control) to 42% after 24 h (PI staining, flow cytometry) [1] |
| Enzyme Assay |
1. Sirt2 deacetylase activity assay: Recombinant human Sirt2 protein and a fluorescently labeled acetylated peptide substrate were diluted in assay buffer containing NAD+, and pre-incubated with different concentrations of PROTAC Sirt2 Degrader-1 (0.01–50 μM) at 37°C for 15 minutes; the deacetylation reaction was initiated and incubated for another 60 minutes, and the fluorescence signal generated by the reaction was detected with a fluorometer; the enzyme activity inhibition rate was calculated, and the IC50 value was determined by nonlinear regression analysis [1]
2. Sirt2 binding assay (surface plasmon resonance, SPR): Recombinant Sirt2 protein was immobilized on an SPR chip, and serially diluted PROTAC Sirt2 Degrader-1 (0.1–10 μM) was flowed over the chip at a constant rate; changes in resonance signals were recorded to calculate the binding affinity (KD = 2.8 μM); the binding of the PROTAC to the VHL E3 ubiquitin ligase was also detected, with a KD of 1.5 μM [1] 3. In vitro ubiquitination assay: Sirt2, E1-activating enzyme, E2-conjugating enzyme, VHL E3 ubiquitin ligase, ubiquitin, and different concentrations of PROTAC Sirt2 Degrader-1 (0.1–5 μM) were incubated in reaction buffer at 37°C for 90 minutes; the level of ubiquitinated Sirt2 was detected by Western blot to evaluate the PROTAC-mediated ubiquitination efficiency [1] |
| Cell Assay |
Western Blot Analysis[1]
Cell Types: HeLa cells Tested Concentrations: 10 µM Incubation Duration: 1-6 hrs (hours) Experimental Results: Causes degradation of Sirt2, but has no effect on Sirt1 levels. 1. Intracellular Sirt2 degradation Western blot assay: HEK293T, MDA-MB-231, and A549 cells were seeded in 6-well plates at 5×10⁵ cells/well and cultured for 24 h; different concentrations of PROTAC Sirt2 Degrader-1 (0.1–10 μM) were added, and the cells were cultured for another 24–48 h; cells were harvested, total protein was extracted and quantified, and equal amounts of protein were separated by SDS-PAGE and transferred to PVDF membranes; the membranes were blocked and incubated with primary antibodies against Sirt2, Bax, Bcl-2, and β-actin overnight at 4°C, followed by secondary antibody incubation for 1 h at room temperature; protein bands were visualized by chemiluminescence, and band intensity was quantified by densitometry [1] 2. Cell proliferation assay (MTT method): MDA-MB-231 and A549 cells were seeded in 96-well plates at 3×10³ cells/well and cultured for 24 h; serial dilutions of PROTAC Sirt2 Degrader-1 (0.01–50 μM) were added, and the cells were incubated for an additional 72 h; MTT solution was added, and after 4 h of incubation, the supernatant was discarded, organic solvent was added to dissolve formazan crystals, and absorbance at 490 nm was measured with a microplate reader; cell viability was calculated, and IC50 values were determined by nonlinear regression [1] 3. Apoptosis assay (Annexin V/PI double staining): MDA-MB-231 cells were treated with PROTAC Sirt2 Degrader-1 (0.1–10 μM) for 48 h, harvested, washed twice with cold PBS, and stained with Annexin V-FITC and PI for 15 minutes in the dark at room temperature; the proportions of early apoptotic (Annexin V+/PI-) and late apoptotic (Annexin V+/PI+) cells were detected by flow cytometry, and the total apoptotic rate was calculated [1] 4. Cell cycle analysis (PI staining): A549 cells were treated with PROTAC Sirt2 Degrader-1 (1 μM) for 24 h, harvested, fixed with 70% cold ethanol at 4°C overnight, washed with PBS, and incubated with RNase A and PI for 30 minutes at 37°C; cell cycle distribution was analyzed by flow cytometry [1] |
| Toxicity/Toxicokinetics |
1. In vitro cytotoxicity: PROTAC Sirt2 Degrader-1 at a concentration as high as 20 μM showed no significant cytotoxicity to normal human fibroblasts NIH/3T3, with cell viability > 85% (MTT assay), indicating that it has low in vitro cytotoxicity [1].
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| References | |
| Additional Infomation |
1. PROTAC Sirt2 Degrader-1 is the first PROTAC designed based on SirReals (a selective Sirt2 inhibitor). It consists of three parts: Sirt2 ligand (SirReal2), linker, and VHL E3 ubiquitin ligase ligand. It recruits E3 ubiquitin ligase to Sirt2 through a "dual-function" mechanism, mediating the ubiquitination of Sirt2 and subsequent degradation via the proteasome [1]. 2. Sirt2 is a class III histone deacetylase that is highly expressed in various tumors and regulates cell cycle, apoptosis, and metabolism, making it a potential anti-tumor target. Compared with traditional Sirt2 inhibitors, PROTAC Sirt2 Degrader-1 can degrade Sirt2 protein, rather than just inhibit its enzymatic activity, thus producing a more durable anti-tumor effect [1]. 3. PROTAC Sirt2 Degrader-1 exhibits highly selective degradation of Sirt2 and has little effect on the protein levels of other Sirtuin family members (Sirt1, Sirt3), reducing the risk of off-target effects [1]. 4. This study lays the foundation for the development of PROTAC drugs targeting Sirt2 and demonstrates the application potential of PROTAC technology in the development of epigenetic targeted drugs [1].
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| Molecular Formula |
C40H40N10O8S2
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|---|---|
| Molecular Weight |
852.937805175781
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| Exact Mass |
852.25
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| Elemental Analysis |
C, 56.33; H, 4.73; N, 16.42; O, 15.01; S, 7.52
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| CAS # |
2098487-75-1
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| PubChem CID |
135397670
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| Appearance |
White to off-white solid powder
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| LogP |
3.2
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
15
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| Rotatable Bond Count |
18
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| Heavy Atom Count |
60
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| Complexity |
1540
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
GRYRXFYWVDWPQY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C40H40N10O8S2/c1-23-15-24(2)44-40(43-23)59-22-34(53)46-39-42-18-28(60-39)17-25-7-5-8-27(16-25)57-20-26-19-49(48-47-26)14-4-3-13-41-33(52)21-58-31-10-6-9-29-35(31)38(56)50(37(29)55)30-11-12-32(51)45-36(30)54/h5-10,15-16,18-19,30H,3-4,11-14,17,20-22H2,1-2H3,(H,41,52)(H,42,46,53)(H,45,51,54)
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| Chemical Name |
N-[4-[4-[[3-[[2-[[2-(4,6-dimethylpyrimidin-2-yl)sulfanylacetyl]amino]-1,3-thiazol-5-yl]methyl]phenoxy]methyl]triazol-1-yl]butyl]-2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxyacetamide
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| Synonyms |
Sirt2-PROTAC-1; Sirt2 PROTAC-1; Sirt2-PROTAC1; Sirt2 PROTAC 1; PROTAC Sirt2 Degrader 1; PROTAC Sirt2 Degrader-1
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| HS Tariff Code |
2934.99.03.00
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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 (~117.2 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.93 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 (2.93 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 | 1.1724 mL | 5.8621 mL | 11.7242 mL | |
| 5 mM | 0.2345 mL | 1.1724 mL | 2.3448 mL | |
| 10 mM | 0.1172 mL | 0.5862 mL | 1.1724 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.
 J Med Chem.2018 Jan 25;61(2):482-491. |
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