| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
| Targets |
Tim9-Tim10 complex (a chaperone complex of the mitochondrial inner membrane TIM22 translocation pathway)[1]
Identified via chemical-genetic synthetic lethality screening; selective against the tim10-1 mutant with an MIC50 of approximately 1 μM.[1] |
|---|---|
| ln Vitro |
- Yeast Growth Inhibition (MIC50): In YPD glucose medium, MitoBloCK-1 exhibits an MIC50 of approximately 1 μM against the tim10-1 mutant; approximately 2 μM against the tim10-73 mutant; and approximately 11.34 μM against the tim9-3 mutant. The MIC50 against the wild-type TIM10 strain is >200 μM.[1]
- Inhibition of Mitochondrial Protein Import: In mitochondria isolated from the tim10-1 tim9S suppressor strain, MitoBloCK-1 inhibits the import of TIM22 pathway substrates in a concentration-dependent manner. AAC import is markedly decreased at 1 μM or higher concentrations. The import of the phosphate carrier (PiC), Tom40, Tim22, and Tafazzin is also inhibited. However, MitoBloCK-1 does not impair the import of TIM23 pathway substrates (e.g., Su9-DHFR, cytochrome b2-DHFR, Hsp60) or Mia40/Erv1 pathway substrates (e.g., Tim9, Tim10, Mia40).[1] - Non-specific Effects on Mitochondrial Function: MitoBloCK-1 does not affect mitochondrial respiration (at 25 μM) or membrane potential (Δψ), nor does it cause non-specific permeabilization of mitochondrial membranes or release of matrix, inner membrane, intermembrane space, or outer membrane proteins. It does not alter the steady-state stability of the Tim9-Tim10 complex.[1] - Activity in Mammalian Cells: Treatment of mammalian cells with 25 μM and 50 μM MitoBloCK-1 results in a significant, dose-responsive decrease in cell viability.[1] - Effect on Isolated Mouse Liver Mitochondria: In isolated mouse liver mitochondria, 25 μM MitoBloCK-1 inhibits the import of AAC but does not affect the import of TIM23 pathway substrates (e.g., Su9-DHFR and Hsp60).[1] |
| Enzyme Assay |
- Yeast Growth Inhibition (MIC50): In YPD glucose medium, MitoBloCK-1 exhibits an MIC50 of approximately 1 μM against the tim10-1 mutant; approximately 2 μM against the tim10-73 mutant; and approximately 11.34 μM against the tim9-3 mutant. The MIC50 against the wild-type TIM10 strain is >200 μM.[1]
- Inhibition of Mitochondrial Protein Import: In mitochondria isolated from the tim10-1 tim9S suppressor strain, MitoBloCK-1 inhibits the import of TIM22 pathway substrates in a concentration-dependent manner. AAC import is markedly decreased at 1 μM or higher concentrations. The import of the phosphate carrier (PiC), Tom40, Tim22, and Tafazzin is also inhibited. However, MitoBloCK-1 does not impair the import of TIM23 pathway substrates (e.g., Su9-DHFR, cytochrome b2-DHFR, Hsp60) or Mia40/Erv1 pathway substrates (e.g., Tim9, Tim10, Mia40).[1] - Non-specific Effects on Mitochondrial Function: MitoBloCK-1 does not affect mitochondrial respiration (at 25 μM) or membrane potential (Δψ), nor does it cause non-specific permeabilization of mitochondrial membranes or release of matrix, inner membrane, intermembrane space, or outer membrane proteins. It does not alter the steady-state stability of the Tim9-Tim10 complex.[1] - Activity in Mammalian Cells: Treatment of mammalian cells with 25 μM and 50 μM MitoBloCK-1 results in a significant, dose-responsive decrease in cell viability.[1] - Effect on Isolated Mouse Liver Mitochondria: In isolated mouse liver mitochondria, 25 μM MitoBloCK-1 inhibits the import of AAC but does not affect the import of TIM23 pathway substrates (e.g., Su9-DHFR and Hsp60).[1] |
| Cell Assay |
- Mammalian Cell Viability Assay (MTT): Mammalian cells were treated with increasing concentrations of MitoBloCK-1 (e.g., 25 μM and 50 μM). After treatment, cell viability was measured using the MTT [1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan] assay.[1]
- Protein Import Assay in Isolated Mouse Liver Mitochondria: Mitochondria were isolated from mouse liver. Radiolabeled precursor proteins (e.g., AAC, Su9-DHFR, Hsp60) were incubated with isolated mitochondria in import buffer containing MitoBloCK-1 (25 μM) or vehicle control. Aliquots were withdrawn at specified time points, and protease was added to remove non-imported precursor. Mitochondria were re-isolated by centrifugation, and samples were analyzed by SDS-PAGE and autoradiography.[1] |
| Toxicity/Toxicokinetics |
The study indicates that in yeast, the compound shows selective toxicity against the tim10 mutant, with an MIC50 >200 μM against the wild-type TIM10 strain. In mammalian cells, 25 μM and 50 μM MitoBloCK-1 treatment significantly reduces cell viability in a dose-dependent manner. At 25 μM, the compound does not impair respiration or membrane potential in isolated yeast mitochondria, nor does it cause non-specific membrane permeabilization.[1]
|
| References | |
| Additional Infomation |
Mechanism of Action: MitoBloCK-1 inhibits an early step in the TIM22 protein import pathway. It hinders the binding of the Tim9-Tim10 complex to its substrates, preventing translocating substrates (e.g., AAC) from crossing the mitochondrial outer membrane. Cross-linking and immunoprecipitation experiments show decreased Tim9-AAC cross-linked products in the presence of MitoBloCK-1.[1]
- Substrate Specificity: Using MitoBloCK-1 as a probe, the substrate specificity of the Tim9-Tim10 complex was determined. The compound inhibits the import of Tim22 and Tafazzin but not Tim23, indicating that Tim23 import depends on the Tim8-Tim13 complex rather than the Tim9-Tim10 complex.[1] - Structure-Activity Relationship (SAR): A limited SAR study of MitoBloCK-1 was conducted. Analogs with modifications to the side chain or the tricyclic core structure were tested. Analog D, with a modified thiourea side chain, inhibited AAC import at 50 μM, while analogs A, B, and C showed no inhibitory activity, suggesting that specific properties of the ring structure and side chain are critical for the activity of MitoBloCK-1.[1] - Screening Methodology: MitoBloCK-1 was discovered through a high-throughput screen of a chemical library of approximately 40,000 compounds using the tim10-1 temperature-sensitive yeast mutant at the permissive temperature of 25°C to identify compounds causing synthetic lethality. Counter-screens using wild-type TIM10 and tim10-1 TIM10 rescue strains were performed to confirm specificity.[1] |
| Molecular Formula |
C14H14BRN3O2S
|
|---|---|
| Molecular Weight |
368.248860836029
|
| CAS # |
373370-73-1
|
| Appearance |
Off-white to light yellow solid powder
|
| SMILES |
BrC1C(=C(C=NNC(N)=S)C2=C(C=1)OC1CCCCC=12)O
|
| Synonyms |
MitoBloCK1; MB-1; MitoBloCK-1; 373370-73-1; MLS000777121; CHEMBL3197716; SCHEMBL13465732;
|
| 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 | 2.7155 mL | 13.5777 mL | 27.1555 mL | |
| 5 mM | 0.5431 mL | 2.7155 mL | 5.4311 mL | |
| 10 mM | 0.2716 mL | 1.3578 mL | 2.7155 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
