AUTAC4 is an autophagy-targeting chimera (AUTAC) that targets mitochondria. AUTAC4 downregulates cytoplasmic proteins and promotes targeted mitochondrial turnover through mitophagy.

Physicochemical Properties

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)

Biological Activity

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].

References

[1]. AUTACs: Cargo-Specific Degraders Using Selective Autophagy. Mol Cell. 2019 Dec 5;76(5):797-810.e10.

Solubility Data

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. 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. 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 corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

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.