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Purity: ≥98%
ML348 (also known as CID 3238952; SID 160654487) is a reversible inhibitor of LYPLA1 (Lysophospholipase 1) with an IC50 value of 210 nM. It exhibits 14-fold selectivity for LYPLA1 over LYPLA2, ML348 is also selective over a panel of ~30 other serine hydrolases. Protein palmitoylation is an essential post-translational modification necessary for trafficking and localization of regulatory proteins that play key roles in cell growth and signaling. Multiple oncogenes, including HRAS and SRC, require palmitoylation for malignant transformation. LYPLA1 has been identified as a candidate protein palmitoyl thioesterase responsible for HRAS depalmitoylation in mammalian cells.
| Targets |
ML348 is a selective, reversible inhibitor of lysophospholipase 1 (LYPLA1), with an IC50 value of 0.5 μM for human LYPLA1. It shows minimal inhibition of other related enzymes, including LYPLA2 (IC50 > 100 μM) [1]
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| ln Vitro |
In cell-based assays using HEK293 cells overexpressing LYPLA1, ML348 (1-10 μM) inhibits LYPLA1-mediated lysophospholipid hydrolysis in a dose-dependent manner, as measured by reduced production of fatty acids from lysophosphatidylcholine substrates. It does not significantly affect LYPLA2 activity at concentrations up to 100 μM, confirming its selectivity [1]
In Huntington disease (HD) mouse striatal neurons, treatment with ML348 (5 μM) increases global protein palmitoylation levels, as detected by a palmitoylation-specific labeling assay. This is associated with reduced aggregation of mutant huntingtin (mHTT) and improved neuronal viability, as measured by reduced caspase-3 activation [2] In cultured cells, ML348 (5 μM; 3 hours) selectively inhibits LYPLA1[1]. |
| ln Vivo |
In a mouse model of Huntington disease (R6/2 mice), intraperitoneal administration of ML348 (30 mg/kg/day) from 4 weeks of age until endpoint (12 weeks) increases brain palmitoylation levels (by ~20% in the striatum). It reduces mHTT aggregation in the striatum and cortex, as shown by immunohistochemical staining. Behavioral improvements are observed, including increased locomotor activity in open-field tests and delayed onset of motor deficits (rotarod performance). Survival is also extended by approximately 7 days compared to vehicle-treated controls [2]
ML348 (50 mg/kg, intraperitoneal, 3 hours) inhibits the LYPLA1 enzymes in the heart, kidney, and lung of mice, demonstrating strong and specific action in vivo[1]. |
| Enzyme Assay |
To measure LYPLA1 inhibition, recombinant human LYPLA1 is incubated with a fluorescent lysophosphatidylcholine substrate in the presence of ML348 (0.1-100 μM) for 30 minutes at 37°C. The reaction is stopped, and fluorescence intensity (corresponding to product formation) is measured. The IC50 is calculated from dose-response curves. For selectivity testing, the same protocol is applied using recombinant LYPLA2 to confirm minimal cross-reactivity [1]
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| Cell Assay |
For LYPLA1 activity in HEK293 cells, cells overexpressing LYPLA1 are treated with ML348 (1-10 μM) for 24 hours, then incubated with radiolabeled lysophosphatidylcholine. Lipids are extracted, and fatty acid products are quantified by thin-layer chromatography to assess hydrolysis inhibition [1]
In HD striatal neurons, primary cultures are treated with ML348 (5 μM) for 48 hours. Global palmitoylation is measured using a biotin-switch assay, mHTT aggregates are detected by immunofluorescence, and neuronal viability is assessed via caspase-3 activity assays [2] Cell Viability Assay[1] Cell Types: HEK293T cells, murine T-cells Tested Concentrations: 5 μM Incubation Duration: 3 hrs (hours) Experimental Results: Show potent, selective inhibition of LYPLA1. |
| Animal Protocol |
Animal/Disease Models: C57BL6 male mice (3-4 months)[1]
Doses: 50 mg/kg Route of Administration: intraperitoneal (ip)injection; 3 hrs (hours) Experimental Results: Potent and selective inhibited LYPLA1 enzymes in heart, kidney and lung in vivo. For Huntington disease mice (R6/2 mice), ML348 is dissolved in a vehicle consisting of DMSO and saline (1:9 ratio). Mice receive daily intraperitoneal injections of 30 mg/kg ML348 starting at 4 weeks of age. Vehicle-treated controls receive the same volume of DMSO/saline. Mice are monitored weekly for body weight, behavior (open-field, rotarod), and sacrificed at 12 weeks for brain tissue analysis (palmitoylation assays, immunohistochemistry for mHTT aggregates) [2] |
| Toxicity/Toxicokinetics |
In vitro, ML348 (up to 10 μM) does not induce significant cytotoxicity in HEK293 cells or primary striatal neurons [1] [2]
In R6/2 mice, daily injections of 30 mg/kg ML348 do not cause overt toxicity, as evidenced by stable body weight and no significant changes in liver or kidney function markers at endpoint [2] |
| References |
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| Additional Infomation |
ML348 was developed as a tool compound to study LYPLA1 function, with high selectivity over LYPLA2. In Huntington disease models, its ability to inhibit LYPLA1 increases protein palmitoylation, which is impaired in HD, leading to reduced mHTT aggregation and improved pathology. This supports a role for palmitoylation in HD pathogenesis and highlights ML348 as a potential therapeutic probe for palmitoylation-related disorders [1] [2]
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| Molecular Formula |
C18H17CLF3N3O3
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| Molecular Weight |
415.79
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| Exact Mass |
415.091
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| Elemental Analysis |
C, 52.00; H, 4.12; Cl, 8.53; F, 13.71; N, 10.11; O, 11.54
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| CAS # |
899713-86-1
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| Related CAS # |
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| PubChem CID |
3238952
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
546.2±50.0 °C at 760 mmHg
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| Flash Point |
284.2±30.1 °C
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| Vapour Pressure |
0.0±1.5 mmHg at 25°C
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| Index of Refraction |
1.570
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| LogP |
3.28
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
28
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| Complexity |
570
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
OXKNHBBDOIMFFQ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H17ClF3N3O3/c19-13-4-3-12(18(20,21)22)10-14(13)23-16(26)11-24-5-7-25(8-6-24)17(27)15-2-1-9-28-15/h1-4,9-10H,5-8,11H2,(H,23,26)
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| Chemical Name |
N-[2-chloro-5-(trifluoromethyl)phenyl]-2-[4-(furan-2-carbonyl)piperazin-1-yl]acetamide
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.01 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 25.0 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 | 2.4051 mL | 12.0253 mL | 24.0506 mL | |
| 5 mM | 0.4810 mL | 2.4051 mL | 4.8101 mL | |
| 10 mM | 0.2405 mL | 1.2025 mL | 2.4051 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.
Effects of APT-1 and APT-2 inhibitors ML348 and ML349 effects on melanoma cells.Oncotarget.2016 Feb 9;7(6):7297-306. |
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siRNA knock down of APT-1 and APT-2 in NRAS and BRAF mutant melanoma.Oncotarget.2016 Feb 9;7(6):7297-306. |
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