Thiolutin (formerly Acetopyrrothin and NSC-3927), is a potent and naturally occusinrg disulfide-containing antibiotic and anti-angiogenic agent isolated from Streptomyces, it acts as a zinc chelator that inhibits the Rpn11 and other JAMM metalloproteases, also inhibits the JAMM metalloproteases Csn5, Associated-molecule-with-the-SH3-Domain-of-STAM (AMSH) and Brcc36. Thiolutin is a potent and selective inhibitor of endothelial cell adhesion accompanied by rapid induction of Heat-shock protein beta-1 (Hsp27) phosphorylation.

Physicochemical Properties

Molecular Formula	C8H8N2O2S2
Molecular Weight	228.29132
Exact Mass	228.002
Elemental Analysis	C, 42.09; H, 3.53; N, 12.27; O, 14.02; S, 28.09
CAS #	87-11-6
PubChem CID	6870
Appearance	Light yellow to yellow solid powder
Density	1.6±0.1 g/cm3
Boiling Point	478.6±45.0 °C at 760 mmHg
Melting Point	273-276℃
Flash Point	243.3±28.7 °C
Vapour Pressure	0.0±1.2 mmHg at 25°C
Index of Refraction	1.721
LogP	0.99
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	1
Heavy Atom Count	14
Complexity	387
Defined Atom Stereocenter Count	0
SMILES	O=C(C)NC1=C2C(=CSS2)N(C)C1=O
InChi Key	MHMRAFONCSQAIA-UHFFFAOYSA-N
InChi Code	InChI=1S/C8H8N2O2S2/c1-4(11)9-6-7-5(3-13-14-7)10(2)8(6)12/h3H,1-2H3,(H,9,11)
Chemical Name	N-(4-methyl-5-oxo-4,5-dihydro-[1,2]dithiolo[4,3-b]pyrrol-6-yl)acetamide
Synonyms	Thiolutin; Acetopyrrothin; NSC 3927; NSC-3927; NSC3927
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)

Biological Activity

References

[1]. Thiolutin is a zinc chelator that inhibits the Rpn11 and other JAMM metalloproteases. Nat Chem Biol. 2017 Jul;13(7):709-714. [2]. Thiolutin inhibits endothelial cell adhesion by perturbing Hsp27 interactions with components of the actin and intermediate filament cytoskeleton. Cell Stress Chaperones. 2010 Mar;15(2):165-81.

Additional Infomation

Thiolutin is a dithiolopyrrolone antibiotic that is 4,5-dihydro[1,2]dithiolo[4,3-b]pyrrole in which the hydrogens at positions 4,5 and 6 have been replaced by methyl, oxo and acetamido groups, respectively. It is a potent inhibitor of RNA polymerases, inhibits the angiogenesis of human umbilical vein endothelial cells, and also inhibits JAMM metalloproteases. It has a role as an angiogenesis inhibitor, a chelator, a protein synthesis inhibitor, an EC 2.7.7.6 (RNA polymerase) inhibitor, an antibacterial agent, a toxin, a marine metabolite, a bacterial metabolite, an antifungal agent and an antineoplastic agent. It is a dithiolopyrrolone antibiotic and a member of acetamides. It is functionally related to a holomycin. Thiolutin has been reported in Streptomyces celluloflavus, Saccharothrix algeriensis, and Streptomyces kasugaensis with data available.

Solubility Data

Solubility (In Vitro)

DMSO : ~6.25 mg/mL (~27.38 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 1.25 mg/mL (5.48 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 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: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: 1.25 mg/mL (5.48 mM)  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	4.3804 mL	21.9020 mL	43.8039 mL
	5 mM	0.8761 mL	4.3804 mL	8.7608 mL
	10 mM	0.4380 mL	2.1902 mL	4.3804 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.