| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Heat - shock protein 70 (Hsp70) and Bcl - 2 interacting mediator of cell death (Bim). S1g - 10 is a specific inhibitor of the Hsp70 - Bim protein - protein interaction.
Heat Shock Protein 70 (Hsp70) and Bim protein-protein interaction (PPI). S1g-10 selectively inhibits the Hsp70-Bim PPI by competitively binding to the NBD domain of Hsp70. In fluorescence polarization assays, its inhibition constant (Ki) is 0.25 μmol/L. [1] |
|---|---|
| ln Vitro |
In OM45 - GFP processing assay, S1g - 10 can interfere with the interaction between Hsp70 and Bim. By using Bax/Bak double - knockout MEF cells, it was found that S1g - 10 could affect the process of mitochondrial outer membrane permeabilization (MOMP), which is related to the apoptosis pathway. The results showed that S1g - 10 could inhibit the function of Hsp70 - Bim interaction, thus affecting the related apoptosis and mitophagy processes.
Inhibition of Hsp70-Bim PPI: In fluorescence polarization assays, S1g-10 competitively inhibits the binding between Hsp70 and the BimBH3 peptide with a Ki value of 0.25 μmol/L, representing approximately a 10-fold improvement in potency compared to the lead compound S1g-2 (Ki = 2.6 μmol/L). [1] Inhibition of ATPase Activity: S1g-10 selectively inhibits Bim-stimulated Hsp70 ATPase activity with a half-maximal inhibitory concentration (IC50) of 15.8 μmol/L, while having no effect on the basal ATPase activity of Hsp70. [1] Induction of Apoptosis: In chronic myeloid leukemia (CML) cell lines, S1g-10 induces apoptosis with EC50 values of 0.62 μmol/L in BV173, 1.3 μmol/L in K562, and 0.34 μmol/L in TKI-resistant K562-R3 cells. It shows no obvious toxicity towards normal HEK293 and BaF3 cells even at 15 μmol/L. [1] Target Specificity Validation: Co-immunoprecipitation (Co-IP) experiments in BV173 cells confirm that S1g-10 (1-10 μmol/L) dose-dependently disrupts the Hsp70-Bim interaction without affecting the Hsp70-Bag3 interaction, demonstrating its specific targeting of the Hsp70-Bim PPI. [1] Impact on Downstream Signaling: Treatment of K562 cells with S1g-10 (2 μmol/L) for 12 hours significantly reduces the protein expression and phosphorylation levels of AKT and EIF4e. [1] Target Dependency Validation: In K562 or BV173 cells stably transduced with Hsp70 or Bim shRNA, S1g-10 (2 μmol/L)-induced apoptosis is significantly attenuated, confirming that its anti-tumor activity depends on the Hsp70-Bim PPI. [1] |
| ln Vivo |
Antitumor Efficacy: In nude mice bearing xenografts of BCR-ABL-independent imatinib-resistant CML cells (K562-R1), intraperitoneal administration of S1g-10 (2 mg/kg, once daily for 14 days) significantly inhibits tumor growth with a T/C ratio of 33.51%, and has negligible effects on mouse body weight. [1]
Target Engagement Validation: Tumors excised from S1g-10-treated mice show disrupted Hsp70-Bim interaction by Co-IP, along with decreased protein levels of AKT and eIF4E, confirming target engagement in vivo. [1] |
| Enzyme Assay |
Fluorescence Polarization Assay for Ki Determination: A fluorescence polarization competition binding assay was performed using a Fam-labeled BimBH3 peptide and recombinant Hsp70 NBD domain (residues 1-383). Increasing concentrations of S1g-10 were incubated with Hsp70 and the Fam-BimBH3 peptide. The change in fluorescence polarization was measured to calculate the Ki value for inhibition of the Hsp70-Bim interaction. VER-155008 was used as a negative control. [1]
Single-Turnover ATPase Activity Assay: The inhibitory effect of S1g-10 on Bim-stimulated Hsp70 ATPase activity was determined in reaction mixtures containing Hsp70, Bim, and varying compound concentrations. The rate of inorganic phosphate production from ATP hydrolysis was measured. The effect on basal Hsp70 ATPase activity was also assessed to verify selectivity. [1] |
| Cell Assay |
Apoptosis Detection by Flow Cytometry: CML cells (BV173, K562, K562-R3) or normal cells (HEK293, BaF3) were seeded in culture plates and treated with various concentrations of S1g-10 for 24 hours. Cells were harvested, stained with Annexin V-FITC and propidium iodide (PI), and analyzed by flow cytometry to determine the percentage of apoptotic cells for EC50 calculation. [1]
Co-Immunoprecipitation: BV173 cells were treated with varying concentrations (1, 2.5, 5, 10 μmol/L) of S1g-10 for 12 hours and then lysed. Immunoprecipitation was performed using an Hsp70-specific antibody, and the levels of co-precipitated Bim and Bag3 were analyzed by Western blot to assess the compound's effect on Hsp70-Bim and Hsp70-Bag3 PPIs. [1] Western Blot Analysis: K562 cells treated with S1g-10 (2 μmol/L) for 12 hours were lysed. Protein levels of AKT, pAKT, EIF4e, pEIF4e, and PARP cleavage fragments were detected using specific antibodies, with β-actin as a loading control. [1] Gene Knockdown Experiments: K562 or BV173 cells stably expressing Hsp70 shRNA or Bim shRNA were generated. Control (non-specific shRNA) or knockdown cells were treated with S1g-10 (2 μmol/L) for 24 hours. Apoptosis rates were assessed by Annexin V staining and flow cytometry, and PARP cleavage was examined to verify target dependency. [1] Use Bax/Bak double - knockout MEF cells. Seed the cells in an appropriate culture medium, and after culturing to the appropriate density, add different concentrations of S1g - 10 solution. Incubate the cells for a certain period of time, and then detect the change of mitochondrial outer membrane permeabilization (MOMP) through relevant methods, such as observing the morphological changes of mitochondria or detecting the release of related proteins. |
| Animal Protocol |
In Vivo Efficacy Study: K562-R1 cells (1 × 10⁶) were subcutaneously implanted into the right flank of sublethally irradiated nude mice. When tumors reached 50-60 mm³, mice were randomized into vehicle control and treatment groups (n=5 per group). S1g-10 was administered intraperitoneally at 2 mg/kg once daily for 14 days. The vehicle was 50% DMSO/PBS. Body weights were measured every 3 days. Mice were sacrificed on day 15, and tumors were excised and weighed. [1]
In Vivo Pharmacokinetic Study: Sprague-Dawley (SD) rats (n=3 per group, 200-250 g) were administered a single intraperitoneal dose of S1g-10 (3 mg/kg). Blood samples were collected at 0, 0.5, 1, 1.5, 2, 4, 6, 8, 12, and 24 hours post-dose. Plasma was separated, and drug concentrations were analyzed by LC-MS/MS to calculate pharmacokinetic parameters. [1] |
| ADME/Pharmacokinetics |
Pharmacokinetic Parameters: In SD rats following a single intraperitoneal dose of S1g-10 (3 mg/kg), the half-life (T1/2) was 5.2 hours, and clearance (CL) was 4.1 mL/min/kg. The maximum plasma concentration (Cmax) was 2.3 μg/mL (approximately 5.3 μmol/L), and the area under the curve (AUC) was 729.1 min·μg/mL (approximately 28.0 μmol·h). [1]
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| References | |
| Additional Infomation |
Background Information: S1g-10 is a small molecule inhibitor. The interaction between Hsp70 and Bim is involved in apoptosis and mitophagy. Research on S1g-10 helps to explore the role of yeast Bxi1/Ybh3 in regulating mitophagy and apoptosis, and reveals a conserved ubiquitin-associated mitophagy pathway in yeast. Furthermore, studies have shown that Ybh3 is multifunctional, possessing the functions of both BH3-domain-only proteins and multi-domain Bax proteins.
|
| Molecular Formula |
C26H31N3OS
|
|---|---|
| Molecular Weight |
433.608844995499
|
| Exact Mass |
433.2187837
|
| CAS # |
3032432-71-3
|
| PubChem CID |
169494198
|
| Appearance |
Light yellow to yellow solid at room temperature
|
| LogP |
6.5
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
31
|
| Complexity |
715
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CCN(CCNC1C2=CC=C(SC3CCCCC3)C3=C2C(=CC=C3)C(=O)C=1C#N)CC
|
| InChi Key |
CFZOZLLGYXBIOC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H31N3OS/c1-3-29(4-2)16-15-28-25-20-13-14-23(31-18-9-6-5-7-10-18)19-11-8-12-21(24(19)20)26(30)22(25)17-27/h8,11-14,18,28H,3-7,9-10,15-16H2,1-2H3
|
| Chemical Name |
6-cyclohexylsulfanyl-3-[2-(diethylamino)ethylamino]-1-oxophenalene-2-carbonitrile
|
| Synonyms |
S1g-10; Hsp70-Bim Inhibitor S1g-10; CHEMBL5397919; 6-(Cyclohexylthio)-3-((2-(diethylamino)ethyl)amino)-1-oxo-1H-phenalene-2-carbonitrile
|
| 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) |
DMSO: ~25 mg/mL (57.7 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3062 mL | 11.5311 mL | 23.0622 mL | |
| 5 mM | 0.4612 mL | 2.3062 mL | 4.6124 mL | |
| 10 mM | 0.2306 mL | 1.1531 mL | 2.3062 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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