| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
ML349 (ML-349) is a novel, potent, reversible, highly selective and specific acyl protein thioesterase 2 (APT-2) inhibitor with a Ki of 120 nM. ML349 is also an inhibitor of LYPLA2 with an IC50 of 144 nM. Although ML349 is highly selective within the serine hydrolase enzyme family, it could still interact with other cellular targets. In human cell lysates, biotinylated-ML349 enriches a recurring set of proteins, including metabolite kinases and flavin-dependent oxidoreductases that are potentially enhanced by avidity-driven multimeric interactions. ML349 achieves target engagement and hydrolase selectivity in living mice.
| Targets |
Ki: 120 nM (APT-2)[1] IC50: 144 nM (LYPLA2)[2]
ML349 inhibits acyl protein thioesterase 1 and 2 (APT-1 and APT-2) with a Kis of >10000 and 120±20 nM, in that order[1]. Additionally, ML349 inhibits LYPLA1 and LYPLA2, with IC50 values of > 3000 and 144 nM, respectively[2]. ML348 and ML349 cause a slight activation of AKT in NRAS mutant cells, but they do not reduce cell viability[3]. |
|---|---|
| ln Vitro |
ML349 inhibits acyl protein thioesterase 1 and 2 (APT-1 and APT-2) with a Kis of >10000 and 120±20 nM, in that order[1]. Additionally, ML349 inhibits LYPLA1 and LYPLA2, with IC50 values of > 3000 and 144 nM, respectively[2]. ML348 and ML349 cause a slight activation of AKT in NRAS mutant cells, but they do not reduce cell viability[3].
|
| Enzyme Assay |
Steady-state kinetic assays were performed to determine inhibition constants (Ki). The fluorogenic substrate resorufin acetate (ResOAc) was used. Reactions were conducted in assay buffer, and hydrolysis was monitored by measuring the increase in fluorescence (excitation/emission) over time. Initial velocities were determined, and data were fit to appropriate inhibition models to calculate Ki values for ML349 and its analogs against wild-type and mutant APT2 enzymes [1].
Fluorescence polarization assays were conducted to measure direct binding of the ML349-FL conjugate to APT2. Increasing concentrations of APT2 were incubated with a fixed concentration of ML349-FL. Polarization values were measured, and data were fit to a binding isotherm to determine the dissociation constant (Kd). Competition experiments were performed by pre-incubating APT2 with unlabeled inhibitors (e.g., ML349, HDFP, or analogs) before adding ML349-FL to assess displacement [1]. Acyl-binding assays utilized the environmentally sensitive fluorophore BODIPY-FL-C16. A sub-micellar concentration of BODIPY-FL-C16 was incubated with increasing concentrations of APT1 or APT2. The increase in fluorescence intensity upon binding to the hydrophobic pocket was monitored. Saturation curves were generated to determine the Kd for lipid binding. For competition, enzymes were pre-incubated with inhibitors like ML349 before adding BODIPY-FL-C16 [1] |
| Cell Assay |
Cells are plated in 96-well plates with a density of 4000 to 8000 cells per well and incubated for 24 h at 37°C with 5% CO2. Then cells are treated with increasing drug (including ML349) concentrations and their combinations. Cell viability is measured with the cell viability assay according to the manufacturers protocol. |
| References | |
| Additional Infomation |
ML349 is a piperazine amide inhibitor containing a thiocyanate sulfone heterocycle in its structure [1]. Its molecular mechanism involves occupying hydrophobic acyl-binding channels near the catalytic triplet (Ser122, His210, Asp176) in APT2, thereby blocking substrate entry [1]. Unlike classic transition state analogs, the sulfonyl group of ML349 does not directly coordinate with the catalytic residues, but rather forms an indirect, water-mediated hydrogen bond network with the oxygen anion pore (the main chain amides of Gln123 and Leu33) and the catalytic histidine (His210) [1]. The selectivity of the inhibitor is influenced by the different binding environments created by β5-α2 ring residues (e.g., Leu78, Ala85, Pro86 in APT2) and G3 helices (e.g., His152, Arg153 in APT2) compared to APT1. Mutations between these residues can alter inhibitor selectivity between the two isoforms [1]. The structure of the APT2-ML349 complex has been resolved by X-ray crystallography at a resolution of 1.64 Å (PDB code: 5SYN) [1].
|
| Molecular Formula |
C23H22N2O4S2
|
|---|---|
| Molecular Weight |
454.561783313751
|
| Exact Mass |
454.102
|
| CAS # |
890819-86-0
|
| PubChem CID |
16193817
|
| Appearance |
White to yellow solid powder
|
| LogP |
3.4
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
31
|
| Complexity |
754
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1(=CC2=C(C3=CC=CC=C3S(=O)(C2)=O)S1)C(N1CCN(C2C=CC(=CC=2)OC)CC1)=O
|
| InChi Key |
JIFSCAIWCCQTRF-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C23H24N2O4S2/c1-29-18-8-6-17(7-9-18)24-10-12-25(13-11-24)23(26)20-14-16-15-31(27,28)21-5-3-2-4-19(21)22(16)30-20/h2-9,14,16,22H,10-13,15H2,1H3
|
| Chemical Name |
(5,5-dioxido-3a,9b-dihydro-4H-thieno[3,2-c]thiochromen-2-yl)(4-(4-methoxyphenyl)piperazin-1-yl)methanone
|
| Synonyms |
ML-349; ML349; ML 349.
|
| 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 : ~20 mg/mL (~44.00 mM)
H2O : < 0.1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2 mg/mL (4.40 mM) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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 20.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1999 mL | 10.9996 mL | 21.9993 mL | |
| 5 mM | 0.4400 mL | 2.1999 mL | 4.3999 mL | |
| 10 mM | 0.2200 mL | 1.1000 mL | 2.1999 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
COA