| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
WEE1; LEB-03-144 targets OTUB1 deubiquitinase (via covalent binding to C23 allosteric site) and WEE1 kinase (via AZD1775 warhead) [1]
|
|---|---|
| ln Vitro |
LEB-03-144 significantly stabilized WEE1 protein levels in HEP3B hepatoma cells. Western blot analysis demonstrated that treatment with LEB-03-144 (concentration not specified) increased WEE1 abundance to levels comparable to the proteasome inhibitor bortezomib (positive control). No stabilization was observed with EN523 (OTUB1 recruiter) or AZD1775 (WEE1 inhibitor) alone, confirming bifunctional dependency [1].
The compound utilized a C3 alkyl linker to conjugate EN523 and AZD1775, which was critical for ternary complex formation. SAR studies indicated that linker length/rigidity influenced efficacy: C3/C5 alkyl linkers (e.g., LEB-03-144, LEB-03-145) stabilized WEE1, while PEG linkers failed [1]. |
| Enzyme Assay |
OTUB1 Engagement Assay**: Recombinant OTUB1 protein was incubated with LEB-03-144 or controls, followed by IA-rhodamine probe labeling. Gel-based ABPP confirmed compound engagement at OTUB1 C23 without inhibiting catalytic activity toward K48-linked di-ubiquitin substrates [1]
|
| Cell Assay |
WEE1 Stabilization Assay: HEP3B cells were treated with LEB-03-144, DMSO (vehicle), EN523, AZD1775, or bortezomib for 24h. Cells were lysed, proteins separated by SDS-PAGE, and WEE1 levels analyzed via Western blot using anti-WEE1 antibody (4936). Band intensity quantification confirmed significant stabilization [1].
|
| References | |
| Additional Infomation |
LEB-03-144 is a deubiquitinating enzyme-targeting chimera (DUBTAC) designed to target and stabilize the WEE1 protein. It links the OTUB1 covalent recruiter EN523 and the WEE1 inhibitor AZD1775 via a C3 alkyl linker. WEE1 is a tumor suppressor that is degraded in cancer via the ubiquitin-proteasome pathway; stabilizing WEE1 may inhibit cancer cell proliferation. The therapeutic significance of stabilizing WEE1 while inhibiting it remains to be explored [1].
The synthesis involves a multi-step organic reaction: first, AZD1775 is linked to a piperazine intermediate, and then coupled to an EN523-modified furanyl propionate via amide coupling. Purity and structure were confirmed by nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS) [1]. |
| Molecular Formula |
C43H51N11O6
|
|---|---|
| Molecular Weight |
817.94
|
| Exact Mass |
817.402378
|
| CAS # |
2858812-89-0
|
| PubChem CID |
162624844
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
3.3
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
13
|
| Rotatable Bond Count |
16
|
| Heavy Atom Count |
60
|
| Complexity |
1520
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC(C)(C1=NC(=CC=C1)N2C3=NC(=NC=C3C(=O)N2CC=C)NC4=CC=C(C=C4)N5CCN(CC5)CCCNC(=O)CCC6=CC=C(O6)N7CCN(CC7=O)C(=O)C=C)O
|
| InChi Key |
RKQZLHHFLGRLEJ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C43H51N11O6/c1-5-20-53-41(58)33-28-45-42(48-40(33)54(53)35-10-7-9-34(47-35)43(3,4)59)46-30-11-13-31(14-12-30)50-24-22-49(23-25-50)21-8-19-44-36(55)17-15-32-16-18-39(60-32)52-27-26-51(29-38(52)57)37(56)6-2/h5-7,9-14,16,18,28,59H,1-2,8,15,17,19-27,29H2,3-4H3,(H,44,55)(H,45,46,48)
|
| Chemical Name |
N-[3-[4-[4-[[1-[6-(2-hydroxypropan-2-yl)pyridin-2-yl]-3-oxo-2-prop-2-enylpyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]piperazin-1-yl]propyl]-3-[5-(2-oxo-4-prop-2-enoylpiperazin-1-yl)furan-2-yl]propanamide
|
| Synonyms |
LEB-03-144; LEB03-144;
|
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2226 mL | 6.1129 mL | 12.2258 mL | |
| 5 mM | 0.2445 mL | 1.2226 mL | 2.4452 mL | |
| 10 mM | 0.1223 mL | 0.6113 mL | 1.2226 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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