Huib32
Based on 1 Customer Validation
Huib32 is a potent small-molecule inhibitor of USP32 (IC50 = 21.2 nM), exhibiting high selectivity over other closely related deubiquitinating enzymes (DUBs), such as USP8/10/16, UCHL1 and OTUB2. Huib32 reversibly inhibits USP32 by covalently binding to the active site Cys743, which enhances substrate ubiquitination, alters endosomal morphology, and mimics USP32 depletion. Huib32 can be used for breast, ovarian, and lung cancer and Alzheimer's and Parkinson’s diseases research.
For research use only. We do not sell to patients.
- Purity: 98.72%
- Formula: C16H23N5O4S
- Molecular Weight:381.45
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
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USP32 21.2 nM (IC50) |
Huib32 (BB01CA282) (0-50 μM, 6-24 h) selectively inhibits both USP32FL and USP32CD in a dose-dependent manner (from 0.1 μM), with no significant effect on 16 other detected DUBs[1].
Huib32 (0-50 μM, 24 h) demonstrates potent, dose-dependent target engagement of endogenous USP32 in MelJuSo cells (IC50 ~0.1 μM), confirming its cell permeability and cellular on-target activity[1].
Huib32 (10 μM, 4-72 h) induces late endosome dispersion and enhances ubiquitination of RAB7 and other substrates (e.g., RAB6, RAB32, RAB11A/B, TMEM192), confirming its role in membrane trafficking and revealing new potential substrates of USP32[1].
Huib32 (10 μM, 4-72 h) significantly alters the ubiquitinome, enriching and depleting unique Lys-ε-Gly-Gly peptides at 4 h and 72 h, impacting substrates like RAB11A/B and USP32 itself without changing their total protein levels, indicating non-proteolytic ubiquitination[1].
Huib32 (10 μM, 72 h) specifically increased the ubiquitination of GFP-RAB7 WT, but not the GFP-RAB7 2KR mutant, validating its on-target effect against an established USP32 substrate[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:MelJuSo cells
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Concentration:0, 0.1, 0.5, 1, 5, 10, and 50 μM
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Incubation Time:4, 24 and 72 h
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Result:Strongly increased the ubiquitination of RAB7 after 4 h of incubation in MelJuSo cells stably expressing GFP-RAB7, which steadily stayed up to 72 h of incubation.
Approximately 2-fold increase in RAB7 ubiquitination was observed in GFP-RAB7 WT, but not in the ubiquitination-deficient mutant GFP-RAB7 2KR.
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Cell Line:MelJuSo cells
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Concentration:0, 5 and 10 μM
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Incubation Time:72 h
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Result:Induced dispersion and swelling of the late endosomal compartments, a phenotype consistent with USP32 depletion.
Chemical Information
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Appearance Solid
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Molecular Weight 381.45
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Formula C16H23N5O4S
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Color White to off-white
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SMILES
CC1=NOC(C)=C1S(N2CCC(C(N[C@@H]3CCN(C#N)C3)=O)CC2)(=O)=O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Solvent & Solubility
DMSO : 100 mg/mL (262.16 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (6.55 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (6.55 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation
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Data Sheet (274 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
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| DMSO | 1 mM | 2.6216 mL | 13.1079 mL | 26.2158 mL | 65.5394 mL |
| 5 mM | 0.5243 mL | 2.6216 mL | 5.2432 mL | 13.1079 mL | |
| 10 mM | 0.2622 mL | 1.3108 mL | 2.6216 mL | 6.5539 mL | |
| 15 mM | 0.1748 mL | 0.8739 mL | 1.7477 mL | 4.3693 mL | |
| 20 mM | 0.1311 mL | 0.6554 mL | 1.3108 mL | 3.2770 mL | |
| 25 mM | 0.1049 mL | 0.5243 mL | 1.0486 mL | 2.6216 mL | |
| 30 mM | 0.0874 mL | 0.4369 mL | 0.8739 mL | 2.1846 mL | |
| 40 mM | 0.0655 mL | 0.3277 mL | 0.6554 mL | 1.6385 mL | |
| 50 mM | 0.0524 mL | 0.2622 mL | 0.5243 mL | 1.3108 mL | |
| 60 mM | 0.0437 mL | 0.2185 mL | 0.4369 mL | 1.0923 mL | |
| 80 mM | 0.0328 mL | 0.1638 mL | 0.3277 mL | 0.8192 mL | |
| 100 mM | 0.0262 mL | 0.1311 mL | 0.2622 mL | 0.6554 mL |
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