| ln Vitro |
In TR-FRET experiments, VWK147 inhibited the interaction between HSP90β CTD and PPID[1]. In fluorescence polarization experiments, VWK147 (10 μM; 3 h) did not bind to HSP90 NTD[1]. In cell-free luciferase renaturation experiments, VWK147 (25–100 μM; 1 h) inhibited HSP90 molecular chaperone function[1]. In BS3 crosslinking experiments, VWK147 (2–50 μM; 1 h) reduced HSP90α CTD dimers[1]. In cisplatin-sensitive T24 and cisplatin-resistant T24-CR urothelial carcinoma cells, VWK147 (1–10 μM; 6 h) destabilized HSP90 client proteins ULK1, RIPK1, and CDK4[1]. VWK147 (0.1-100 μM; 72 hours) reduced the viability of cisplatin-sensitive (253J, T24) and drug-resistant (253J-CR, T24-CR) urothelial carcinomas, with an IC50 value of approximately 3-5 μM after 72 hours [1]. VWK147 (10 μM; 0-24 hours) induced apoptosis and necrotizing death in T24 and T24-CR urothelial carcinomas within 24 hours [1]. The process by which VWK147 (5 μM; 24 hours) induced death in 253J, 253J-CR, T24 and T24-CR urothelial carcinomas was partially caspase-dependent [1]. VWK147 (1-10 μM; 6 hours) induced PARP1 cleavage in T24 and T24-CR urothelial carcinomas in a concentration-dependent manner [1]. VWK147 (5 μM; 1–24 h) induces time-dependent cleavage of PARP1 in T24 and T24-CR urothelial carcinoma cells within 24 hours [1]. VWK147 (5 μM; 0–24 h) induces caspase-3 activation in cisplatin-sensitive (253J, T24) and drug-resistant (253J-CR, T24-CR) urothelial carcinoma cells within 24 hours [1]. VWK147 (5 μM; 6 h) inhibits autophagy flux in T24 and T24-CR urothelial carcinoma cells, manifested as LC3-II accumulation [1]. VWK147 (5 μM; 4 h) inhibits autophagosome-lysosome fusion in T24 and T24-CR urothelial carcinoma cells stably expressing mRFP-EGFP-rLC3 [1].
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| Cell Assay |
Western Blot Analysis [1]
Cell Types: Cisplatin-sensitive (T24) and drug-resistant (T24-CR) urothelial carcinoma cells Tested Concentrations: 1 μM, 3 μM, 5 μM, 10 μM Incubation Duration: 6 hours Experimental Results: The levels of HSP90 client proteins ULK1, RIPK1, and CDK4 were decreased in T24 and T24-CR cells. The levels of HSP27, HSP40, or HSP70 were not increased. Cell viability assay [1] Cell Types: Cisplatin-sensitive (253J, T24) and drug-resistant (253J-CR, T24-CR) urothelial carcinoma cells Tested Concentrations: 0.1-100 μM Incubation Duration: 72 hours Experimental Results: Cell viability was reduced in all four cell lines. The IC50 values of 253J, 253J-CR, T24 and T24-CR were ~3-5 μM, ~3-5 μM, ~3-5 μM and ~3-5 μM, respectively. Cell viability assay [1] Cell Types: 253J, 253J-CR, T24 and T24-CR urothelial carcinoma cells Tested Concentrations: 5 μM (co-treated with 20 μM Q-VD-OPh ()) Incubation Duration: 24 hours Experimental Results: Q-VD-OPh partially reduced the cell viability reduction mediated by it, but did not completely eliminate it, indicating that it has a partial pro-apoptotic effect.
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| References |
| Molecular Formula |
C32H38N10O7
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|---|---|
| Molecular Weight |
674.71
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| Appearance |
Typically exists as solids at room temperature
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| SMILES |
CCC(N1C(C(NC(C2=NC=C(C(N2CC3=CC=C(C=C3)OC)=O)N)=O)=CN=C1C(NC4=CN=C(N(C4=O)CC(C)C)C(NC)=O)=O)=O)C
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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) |
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
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.4821 mL | 7.4106 mL | 14.8212 mL | |
| 5 mM | 0.2964 mL | 1.4821 mL | 2.9642 mL | |
| 10 mM | 0.1482 mL | 0.7411 mL | 1.4821 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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