| Targets |
USP8 (IC50 = 4.0 μM)
USP8-IN-3 targets ubiquitin-specific protease 8 (USP8), a deubiquitinating enzyme that removes ubiquitin moieties from target proteins. USP8 plays a critical role in various cellular processes, including endosomal sorting, receptor trafficking, and cell signaling. By inhibiting USP8, this compound disrupts these processes, leading to anti-proliferative effects. It also inhibits USP7. |
|---|---|
| ln Vitro |
In vitro, USP8-IN-3 demonstrates potent inhibition of USP8 with an IC50 of 4.0 μM. It inhibits the proliferation of H1957 cells (GI50 = 6.01 μM) and GH3 rat pituitary cells (GI50 = 37.03 μM). Its activity against USP7 suggests a broader mechanism.
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| ln Vivo |
In vivo, USP8-IN-3 is being investigated for its potential to treat cancer and viral infections. Its anti-proliferative activity in cellular models supports its further development as a therapeutic agent. However, specific in vivo efficacy data is not detailed in the available literature.
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| Enzyme Assay |
The inhibitory activity of USP8-IN-3 is assessed using in vitro deubiquitinase assays. Recombinant USP8 is incubated with a ubiquitinated substrate and varying concentrations of the compound. The deubiquitination reaction is measured, and the IC50 is calculated.
|
| Cell Assay |
Western Blot Analysis[1]
Cell Types: / Tested Concentrations: 6.25 μM, 12.5 μM, 25 μM, USP8-IN-3 (6.25-100 μM; 15 min) inhibits the cleavage activity of USP8 on dimeric ubiquitin[1]. Incubation time at 50 μM and 100 μM: 15 minutes; add Di-Ub substrate for an additional hour. Experimental Results: diminished levels of cleaved diubiquitin. The cellular activity of USP8-IN-3 is evaluated in cancer cell lines, such as H1975 (NSCLC) and GH3 cells. Cells are treated with the compound, and its effect on cell proliferation is measured using standard assays. |
| Animal Protocol |
In animal studies, USP8-IN-3 would be administered to mice bearing tumor xenografts to assess its anti-tumor activity. Its effect on tumor growth and pharmacodynamic markers would be evaluated.
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| ADME/Pharmacokinetics |
As a small molecule (MW 425.43), USP8-IN-3 has properties suitable for in vivo studies. Its pharmacokinetic properties would need to be determined for specific applications.
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| Toxicity/Toxicokinetics |
Toxicology data for USP8-IN-3 is limited. As a research compound, its safety profile has not been established in formal toxicology studies. Its use is limited to research applications.
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| References | |
| Additional Infomation |
USP8-IN-3 (CAS: 2477651-10-6) is a potent inhibitor of USP8 and USP7. It is a valuable research tool for studying the role of deubiquitinating enzymes in cancer and viral infections. Its ability to inhibit the proliferation of NSCLC cells highlights its potential relevance for lung cancer research.
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| Exact Mass |
425.11
|
|---|---|
| Elemental Analysis |
C, 50.82; H, 4.26; F, 13.40; N, 16.46; O, 7.52; S, 7.54
|
| CAS # |
2477651-10-6
|
| Related CAS # |
2477651-10-6
|
| PubChem CID |
150116188
|
| Appearance |
Light yellow to yellow solid
|
| LogP |
3.9
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
29
|
| Complexity |
573
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
DYHHEFBWFATDAN-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C18H18F3N5O2S/c19-18(20,21)15-2-1-3-16(24-15)25-10-8-13(9-11-25)23-17(29)22-12-4-6-14(7-5-12)26(27)28/h1-7,13H,8-11H2,(H2,22,23,29)
|
| Chemical Name |
1-(4-nitrophenyl)-3-(1-(6-(trifluoromethyl)pyridin-2-yl)piperidin-4-yl)thiourea
|
| Synonyms |
USP8-IN-3; DUN-51106; DUN 51106; DUN51106
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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)
|
| Solubility (In Vitro) |
DMSO: ~100 mg/mL (~235.1 mM)
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|---|---|
| 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.) |
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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