| 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 |
| Targets |
IC50: 0.57 μM (CRBN)[1]
CRBN (cereblon); ALV2 binds to CRBN with an IC50 of 0.57 uM in a TR-FRET assay. It recruits the E3 ubiquitin ligase substrate receptor cereblon to Helios (IKZF2), leading to the ubiquitination and degradation of Helios. |
|---|---|
| ln Vitro |
In the TR-FRET test, ALV2 (10 nM-100 μM) causes CRBN-Helios dimerization[1]. In Jurkat cells, ALV2 (0.1–10 μM) primarily stimulates Helios degradation[1]. In Jurkat cells, ALV2 (1 μM) pretreatment for 18 hours increases IL-2 secretion[1].
In the TR-FRET assay, ALV2 (10 nM-100 uM) causes CRBN-Helios dimerization. In Jurkat T cells, ALV2 (0.1-10 uM) primarily stimulates Helios degradation. In Jurkat cells, ALV2 (1 uM) pretreatment for 18 hours increases IL-2 secretion, indicating enhanced T cell activation. |
| ln Vivo |
In vitro, ALV2 (100 mg/kg; intraperitoneally twice daily for seven days) selectively degrades Helios[1].
In vivo, ALV2 (100 mg/kg; intraperitoneally twice daily for seven days) selectively degrades Helios in CrbnI391V/I391V mice. It decreases Helios levels in splenic CD4+FoxP3+ regulatory T (Treg) cells without affecting Ikaros levels, demonstrating target specificity. |
| Enzyme Assay |
(1) CRBN binding TR-FRET assay: Mix CRBN protein (Eu-labeled), biotinylated Helios peptide, streptavidin-XL665, and ALV2 (0.1 nM - 100 uM). (2) Incubate at room temperature for 2-3 h. (3) Measure fluorescence at 620 nm (donor) and 665 nm (acceptor). (4) Calculate IC50 from 665/620 ratio; the IC50 for ALV2 is 0.57 uM.
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| Cell Assay |
(1) Seed Jurkat cells (1×10⁵ cells/mL) in 12-well plates. (2) Treat with ALV2 at 0.1, 1, 10 uM for 18 h. (3) For Helios degradation: lyse cells, perform Western blot with anti-Helios antibody (detects IKZF2) and anti-Ikaros (IKZF1) as control. (4) For IL-2 secretion: pre-treat with ALV2 (1 uM, 18 h), then stimulate with anti-CD3/anti-CD28 for 24 h, collect supernatant, measure IL-2 by ELISA.
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| Animal Protocol |
Animal/Disease Models: CrbnI391V/I391V mice
Doses: 100 mg/kg Route of Administration: Ip twice (two times) daily for 7 days Experimental Results: decreased Helios, but not Ikaros, levels in splenic CD4+FoxP3+ Treg cells. (1) Use 6-8 week old CrbnI391V/I391V mice (male and female, 20-25 g). (2) Administer ALV2 intraperitoneally (IP) at 100 mg/kg twice daily for 7 days. (3) Formulation: dissolve in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. (4) On day 7, sacrifice mice, collect spleens, isolate CD4+ T cells. (5) Analyze Helios and Ikaros levels in CD4+FoxP3+ Treg cells by flow cytometry (intracellular staining) and Western blot. (6) Determine percentage of Helios degradation relative to vehicle control. |
| ADME/Pharmacokinetics |
Standard formulation for in vivo: prepare 20-50 mg/mL stock in DMSO, then dilute in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline for IP injection. Solubility in DMSO: ~160 mg/mL (~305.36 mM). LogP: 2.5 (predicted). PK in mice (IP, 100 mg/kg, predicted): Cmax ~ 20-50 uM, t1/2 ~ 2-4 h, AUC ~ 50-100 uM·h, moderate clearance (CL ~ 20-40 mL/min/kg).
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| Toxicity/Toxicokinetics |
For in vitro toxicity: CCK-8 assay on HEK293 cells, IC50 > 50 uM. For in vivo toxicity: MTD studies in CrbnI391V/I391V mice, single IP dose up to 200 mg/kg; repeated dosing at 100 mg/kg IP twice daily for 7 days is well tolerated, with no significant weight loss or mortality. Monitor clinical signs, body weight, and serum chemistry (ALT, AST, BUN, creatinine). The compound is for research use only, not for human use.
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| References | |
| Additional Infomation |
ALV2 is a first-in-class selective Helios (IKZF2) molecular glue degrader that binds to cereblon with an IC50 of 0.57 microM. It specifically degrades Helios without affecting the related family member Ikaros, making it valuable for studying Treg cell biology and immune-oncology. By destabilizing Treg cells, ALV2 may enhance anti-tumor immune responses. This compound is not FDA-approved and is strictly for research use.
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| Molecular Formula |
C26H26CLN5O5
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|---|---|
| Molecular Weight |
523.97
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| Exact Mass |
523.162
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| CAS # |
2438124-95-7
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| PubChem CID |
146628167
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| Appearance |
White to yellow solid powder
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| LogP |
2.5
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
37
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| Complexity |
998
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=C(C=C(C=C1)NC(=O)NC(C)(C)C2=CC(=CC=C2)NC3=CC(=O)N(C3=O)C4CCC(=O)NC4=O)Cl
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| InChi Key |
YKFHBIMJVFQOQL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H26ClN5O5/c1-14-7-8-17(12-18(14)27)29-25(37)31-26(2,3)15-5-4-6-16(11-15)28-19-13-22(34)32(24(19)36)20-9-10-21(33)30-23(20)35/h4-8,11-13,20,28H,9-10H2,1-3H3,(H2,29,31,37)(H,30,33,35)
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| Chemical Name |
1-(3-chloro-4-methylphenyl)-3-[2-[3-[[1-(2,6-dioxopiperidin-3-yl)-2,5-dioxopyrrol-3-yl]amino]phenyl]propan-2-yl]urea
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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 :~160 mg/mL (~305.36 mM)
H2O :< 0.1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 4 mg/mL (7.63 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 40.0 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: 4 mg/mL (7.63 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 40.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9085 mL | 9.5425 mL | 19.0851 mL | |
| 5 mM | 0.3817 mL | 1.9085 mL | 3.8170 mL | |
| 10 mM | 0.1909 mL | 0.9543 mL | 1.9085 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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