| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
The primary target of AUTAC4 is the translocator protein (TSPO) on the outer mitochondrial membrane. TSPO is a mitochondrial protein that is involved in various cellular functions, including cholesterol transport, steroidogenesis, and mitochondrial permeability transition. AUTAC4 contains a 2-phenylindole-3-glyoxyamide ligand that binds to TSPO with high affinity. The compound also contains a p-fluorobenzyl guanine (FBnG) moiety that serves as an autophagy-inducing tag. By linking the TSPO ligand to the FBnG tag via a PEG linker, AUTAC4 recruits the autophagy machinery to mitochondria, promoting their selective degradation through mitophagy.
|
|---|---|
| ln Vitro |
AUTAC4 causes polyubiquitination associated to K63. It took around 8 hours of incubation for K63-linked polyubiquitin to accumulate[1]. AUTAC4 inhibits pro-caspase 3 cleavage in addition to the release of cytochrome c into the cytoplasm. AUTAC4 also maintains the intracellular ATP level following CCCP therapy, indicating that via enhancing mitochondrial quality control through mitophagy acceleration, AUTAC4 restored energy production. Via the stimulation of mitophagy, AUTAC4 protects cells against acute mitochondrial damage[1]. AUTAC4 (10 μM) can induce mitophagy in Detroit 532 cells around 24-72 hours post-treatment using mito-Rosella[1].
The in vitro activity of AUTAC4 has been characterized in cellular models of mitochondrial dysfunction. The compound induces selective mitophagy by tagging mitochondria for degradation via the autophagy-lysosome pathway. AUTAC4 downregulates cytosolic proteins while promoting selective mitochondrial degradation. The compound has been shown to reverse mitochondrial dysfunction through K63-linked ubiquitination. The half-maximal effective concentration (EC50) for mitophagy induction has not been detailed in the available literature, but the compound is described as a potent inducer of selective autophagy. AUTAC4 represents a novel therapeutic strategy for targeting mitochondrial dysfunction. |
| ln Vivo |
No detailed in vivo activity data for AUTAC4 has been published in the available literature. The compound has been primarily characterized in vitro as a tool for studying mitophagy and mitochondrial dysfunction. Further studies would be needed to evaluate the efficacy of AUTAC4 in animal models of mitochondrial diseases, neurodegenerative disorders, or cancer. The ability of AUTAC4 to cross biological barriers and reach target tissues in vivo would need to be assessed.
|
| Enzyme Assay |
The binding of AUTAC4 to TSPO on the outer mitochondrial membrane can be assessed using radioligand binding assays or surface plasmon resonance (SPR). In a typical radioligand binding assay, mitochondrial membranes or cells expressing TSPO are incubated with a radiolabeled TSPO ligand (such as ³H-PK11195) and varying concentrations of AUTAC4. The displacement of the radiolabeled ligand by AUTAC4 is measured, and the binding affinity (Ki) is calculated. The specificity of AUTAC4 for TSPO can be confirmed by using TSPO-knockout cells or by competition with known TSPO ligands.
|
| Cell Assay |
The cellular activity of AUTAC4 is assessed using cell lines or primary cells with mitochondrial dysfunction. Cells are treated with AUTAC4 at varying concentrations for defined periods. The induction of mitophagy is assessed by monitoring the colocalization of mitochondria (labeled with MitoTracker or by immunostaining for mitochondrial proteins) with autophagosomes or lysosomes (labeled with LC3 or LAMP1 antibodies) using confocal microscopy. The degradation of mitochondrial proteins, such as Tom20 or COX IV, is assessed by Western blotting. The effects of AUTAC4 on mitochondrial function are assessed by measuring mitochondrial membrane potential (using JC-1 dye), oxygen consumption rate (using a Seahorse analyzer), or ATP production.
|
| Animal Protocol |
No detailed in vivo animal model data for AUTAC4 has been published in the available literature. As a research tool, AUTAC4 has not been evaluated in animal models to assess its in vivo efficacy or pharmacokinetic properties. Future studies may involve the use of mouse models of mitochondrial diseases or neurodegenerative disorders to assess the therapeutic potential of AUTAC4.
|
| ADME/Pharmacokinetics |
No detailed pharmacokinetic data for AUTAC4 has been published in the available literature. As a heterobifunctional molecule with a PEG linker, AUTAC4 is likely to have distinct pharmacokinetic properties compared to small molecules. The compound’s absorption, distribution, metabolism, and excretion (ADME) properties would need to be characterized in future studies to determine its suitability for in vivo applications. Factors such as oral bioavailability, plasma protein binding, clearance, half-life, and tissue distribution would be important for understanding its pharmacokinetic profile.
|
| Toxicity/Toxicokinetics |
No detailed toxicity data for AUTAC4 has been published in the available literature. As a research compound, AUTAC4 has not been subjected to comprehensive toxicology studies. The mechanism of action, which involves the induction of mitophagy, could have off-target effects if the compound induces autophagy in normal tissues or if the guanine tag triggers immune responses. Further studies would be needed to assess the safety profile of AUTAC4.
|
| References | |
| Additional Infomation |
AUTAC4 is a first-generation autophagy-targeting chimera (AUTAC) designed to induce selective mitophagy. The compound comprises a p-fluorobenzyl guanine (FBnG) moiety joined by a PEG linker to a 2-phenylindole-3-glyoxyamide ligand that binds to the translocator protein (TSPO) on the outer mitochondrial membrane. AUTAC4 promotes the selective degradation of mitochondria through the autophagy-lysosome pathway. The compound is a valuable research tool for studying mitophagy and mitochondrial dysfunction and represents a novel therapeutic strategy for conditions associated with mitochondrial dysfunction.
|
| Molecular Formula |
C43H48FN9O8S
|
|---|---|
| Molecular Weight |
869.96
|
| Exact Mass |
869.333
|
| CAS # |
2267315-04-6
|
| PubChem CID |
137409907
|
| Appearance |
White to light yellow solid powder
|
| LogP |
3
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
12
|
| Rotatable Bond Count |
24
|
| Heavy Atom Count |
62
|
| Complexity |
1520
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
C1=C(F)C=CC(CN2C3N=C(N)NC(=O)C=3N=C2SC[C@H](NC(C)=O)C(=O)NCCCOCCOCCOCCCNC(=O)C(=O)C2=C(C3=CC=CC=C3)NC3=CC=CC=C23)=C1
|
| InChi Key |
IYRHHSZRPVKBSQ-XIFFEERXSA-N
|
| InChi Code |
InChI=1S/C43H48FN9O8S/c1-27(54)48-33(26-62-43-50-36-38(51-42(45)52-40(36)57)53(43)25-28-13-15-30(44)16-14-28)39(56)46-17-7-19-59-21-23-61-24-22-60-20-8-18-47-41(58)37(55)34-31-11-5-6-12-32(31)49-35(34)29-9-3-2-4-10-29/h2-6,9-16,33,49H,7-8,17-26H2,1H3,(H,46,56)(H,47,58)(H,48,54)(H3,45,51,52,57)/t33-/m0/s1
|
| Chemical Name |
(2R)-2-acetamido-3-[[2-amino-9-[(4-fluorophenyl)methyl]-6-oxo-1H-purin-8-yl]sulfanyl]-N-[3-[2-[2-[3-[[2-oxo-2-(2-phenyl-1H-indol-3-yl)acetyl]amino]propoxy]ethoxy]ethoxy]propyl]propanamide
|
| 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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : 170 mg/mL (195.41 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 4.25 mg/mL (4.89 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 42.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 4.25 mg/mL (4.89 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 42.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 4.25 mg/mL (4.89 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.1495 mL | 5.7474 mL | 11.4948 mL | |
| 5 mM | 0.2299 mL | 1.1495 mL | 2.2990 mL | |
| 10 mM | 0.1149 mL | 0.5747 mL | 1.1495 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
